Article

Collagencin, an antibacterial peptide from fish collagen: Activity, structure and interaction dynamics with membrane

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Abstract

In this study, we first report characterization of collagencin, an antimicrobial peptide identified from fish collagen hydrolysate. The peptide completely inhibited the growth of Staphylococcus aureus at 1.88 mM. Although non-toxic up to 470 μM, collagencin was hemolytic at higher concentrations. The secondary structure of collagencin was mainly composed by β-sheet and β-turn as determined by CD measurements and molecular dynamics. The peptide is likely to form β-sheet structure under hydrophobic environments and interacts with both anionic (phosphatidylglycerol) and zwitterionic (phosphoethanolamine and phosphatidylcholine) lipids as shown with CD spectroscopy and molecular dynamics. The peptide formed several hydrogen bonds with both POPG and POPE lipids and remained at membrane–water interface, suggesting that collagencin antibacterial action follows a carpet mechanism. Collagenous fish wastes could be processed by enzymatic hydrolysis and transformed into products of high value having functional or biological properties. Marine collagens are a promising source of antimicrobial peptides with new implications in food safety and human health.

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... It is also the duty of them to safeguard the crayfish against in the marine world, diverse fish pathogens. The antimicrobial peptide astacidin was derived from the freshwater crayfish Pacifastacus leniusculus, and it has a large spectrum of bactericidal potential against both Gram-positive and Gram-negative bacteria (Jiravanichpaisal et al. 2007;Ennaas et al. 2016). In an In vitro study Ennaas et al. (2016) extracted and characterized Collagencin, a bactericidal peptide with good action against multidrug-resistant Staphylococcus aureus (Ennaas et al. 2016). ...
... The antimicrobial peptide astacidin was derived from the freshwater crayfish Pacifastacus leniusculus, and it has a large spectrum of bactericidal potential against both Gram-positive and Gram-negative bacteria (Jiravanichpaisal et al. 2007;Ennaas et al. 2016). In an In vitro study Ennaas et al. (2016) extracted and characterized Collagencin, a bactericidal peptide with good action against multidrug-resistant Staphylococcus aureus (Ennaas et al. 2016). Dermaseptin is a brand-new linear peptide with antimicrobial effects. ...
... The antimicrobial peptide astacidin was derived from the freshwater crayfish Pacifastacus leniusculus, and it has a large spectrum of bactericidal potential against both Gram-positive and Gram-negative bacteria (Jiravanichpaisal et al. 2007;Ennaas et al. 2016). In an In vitro study Ennaas et al. (2016) extracted and characterized Collagencin, a bactericidal peptide with good action against multidrug-resistant Staphylococcus aureus (Ennaas et al. 2016). Dermaseptin is a brand-new linear peptide with antimicrobial effects. ...
Chapter
Infectious diseases are one of the major considerable factors in the rising universal mortality and morbidity rate. The frequency of microbial infections is gaining higher in communities due to the sudden ingress of life-threatening newer pathogens or the acquired emergence of drug-resistant pathogenic microbes. Extensive misuse and overuse in clinical practices led to the development of reduced susceptibility of the drugs to the current armamentarium of antimicrobial agents. Undoubtedly, antimicrobial agents play a remarkable role to support science in order to eradicate microbial infections to a certain level and also help to accelerate the development of newer antimicrobial drugs to combat microbial drug-resistant issues. In the finding of newer drugs, natural sources or herbal medicines have been proved to be excellent resources of antimicrobial compounds in the present time. These potent newer natural derived molecules have been shown to save lives against severe infectious diseases and also reduce the burden of illness in certain clinical situations. Many natural compounds and antibiotics isolated from microbes or microbial strains also face strong challenges due to their ineffectiveness when other strains get developed resistance against such molecules. To eradicate these major issues, scientists and researchers are now well focused on the development of efflux pump inhibitors to combat this emerging seriousness against a wide variety of spectrum by giving life to the ineffective/old antibiotics. The issue we’re answering in this chapter is whether natural sources that contain a wide spectrum of bioactive secondary metabolites can able to confer new antibiotics to fight MDR microbes and novel chemotherapy stimulators to restore old antibiotics to become therapeutically effective. The key aim of this research is to illustrate the possible benefits of a number of naturally derived compounds as well as the role of phytochemicals in the growth of biocompatible therapeutics. This study also focuses on the different effects and efficacy of natural compounds in controlling MDR in bacteria, with the aim of promoting research into unexplored plants in order to find new antibiotics for global health benefits.
... Recently, collagen extracted from fish by-products has been used by the food, biomedical, pharmaceutical, and cosmetic industries because of its properties such as gelling and surface properties, excellent biocompatibility, and other biological characteristics (Peres et al., 2017). However, besides being soluble in water, the low-molecularweight peptides obtained from the hydrolysis of fish collagen usually present major bioavailability and a greater variety of bioactivities than the raw collagen (Hajfathalian, Ghelichi, García, Moltke, & Jacobsen, 2018); including antioxidant, antimicrobial, antibacterial, angiotensinconverting enzyme (ACE-inhibitory), anti-hyperglycemic anti-Alzheimer's and neuroprotective activity (Ennaas et al., 2016;Hajfathalian et al., 2018;Abuine, Rathnayake, & Byun, 2019;Wang, Zhao, Zhao, & Chi, 2020). ...
... Likewise, Trang and Pasuwan (2018) reported the effect of inhibition at 8% (mg/mL) extract powder on E. Coli (0157:H7) registering an inhibition zone of 13-18 mm, whereas the molecular weight of the hydrolysate was 5 kDa. Ennaas et al. (2016) also rereported inhibition on E. Coli (MC4100) by collagencin (MW 1 kDa), a peptide separated and identified with a sequence of GLPGP-LGPAGPK from fish collagen hydrolysate. Results for F2 can be associated with the presence of amino acids with a positive charge (16%), such as lysine, arginine, and histidine, in addition to the hydrophobic amino-acid content (60%) as alanine, leucine, isoleucine, valine, proline, phenylalanine, methionine, serine, tyrosine, threonine, and glycine (Table 2), because amino acids charged positively in peptide sequences penetrate bacterial membranes interacting with the negative charges of the cell wall of phospholipids, promoting cellular lysis (Hajfathalian et al., 2018). ...
Article
Fish by-products are excellent sources of collagen. Acid-soluble collagen (ASC) derived from a mixed by-product of different fish species was hydrolyzed to obtain peptide fractions and evaluate their biological and functional activities. All fractions obtained (F1: ≥30, F2: 10-30, F3: 5-10, F4: 1-5, and F5: ≤1kDa) exhibited antioxidant activity at concentrations of 5, 10, and 15 mg/mL. However, F5 registered the highest reducing power (absorbance 0.366) and hydroxyl-radical-scavenging activity (91%) at 15 mg/mL; whereas the highest DPPH• scavenging activity (81%) was also detected in F5 at 5 mg/mL. The solubility of F1, F2, and F3 was ≥95% at pH 7. The highest foaming capacity (78%), foaming stability (60%), and emulsion stability index (42 min) were registered for F1. However, the highest emulsifying activity index (130 m²/g) was for F3. These results place collagen obtained from a mixed by-product of different fish species as a potential biotechnological alternative for the industry.
... This experimental model provides a good platform to discover the phases of healing through re-epithelization process, granulation tissue angiogenesis and the histological pattern of the cells and extracellular matrix (ECM) [21,22] . Daily assessment of the wound indicates sign of significant differences during the healing process by fibroblasts and has a major role in the proliferative and remodeling phase [25] . Subsequently, the wound maturation phase started after the proliferative phase. ...
... It has been acknowledged that collagen plays an essential role in wound healing process and is one of the structural and functional components of extracellular matrix in the dermis. Collagen is also a good substitute for skin material, and the properties of marine collagens may include antibacterial, antioxidant and anti-skin-aging activities [25,29] . In general, it is believed that collagen is a good biomaterial source for tissue engineering due to its adeptness for cellular-adherence and biological affinity [29] . ...
Article
Fish collagen derived from its skins, scales or bones are known to have many advantages in medical and pharmaceutical applications. This study evaluates the effectiveness of collagen extracted from the scales of local Malaysian Oreochromis niloticus or red Nile tilapia fish, on wound healing in an animal model. 21 male BALB/c mice were divided into three different groups namely the control group, Oreochromis niloticus acid-solubilized collagen extracts treated group; and acriflavine treated group. A full thickness surgical wound of about 1.5 cm were inflicted on the dorsal area of the animal. Treatment with collagen extracts and acriflavine were applied daily on the wounds and measured every day for 9 days. The mice were randomly sacrificed on day 3, 6, and 9, and the skin samples were taken for histological assessment. After 3 days, treatment with collagen extracts and acriflavine recorded an average wound measurement of 7.84±0.32 mm and 7.14±0.32, respectively. On day 9, the progress of wound healing showed significant differences. Mice treated with collagen extracts recorded an average wound measurement of 1.27±0.08 mm, as compared to 3.26±0.05 mm of acriflavine treated, and 3.36±0.18 mm of the control group. The wound assessment showed that collagen extracted from the scales of local Oreochromis niloticus fish produced significant results based on the skin re-epithelialization process and formation of new blood vessels. This finding indicates that collagen extracted from the local Oreochromis niloticus scales possessed effective wound healing properties for surgical wounds as demonstrated on the skin of the animal model.
... These elements differ from collagens derived from farmed land animals (cattle, pigs) and give them different physico-chemical properties due to their amino acid composition. Collagens of marine origin have antibacterial, antioxidant, neuroprotective, and anti-aging properties [48][49][50][51]. In addition, marine peptides are believed to promote skin healing in rats [52]. ...
... Although it is well-known that peptides from dietary supplements can be found in the bloodstream, the concentration of the dietary supplement that actually circulates in the body is unknown. For example, upon oral ingestion of gelatin hydrolysates derived from porcine skin, chicken feet, and cartilage after 12 h of fasting, the peptide form of hydroxyproline increases significantly and reaches a maximum level (20-60 nmol/mL of plasma) after 1-2 h, and then decreases to half the maximum level 4 h after ingestion [48]. The main constituents of food collagen peptides in human serum and plasma have been identified as Pro-Hyp. ...
Article
Full-text available
Articular cartilage experiences mechanical constraints leading to chondral defects that inevitably evolve into osteoarthritis (OA), because cartilage has poor intrinsic repair capacity. Although OA is an incurable degenerative disease, several dietary supplements may help improve OA outcomes. In this study, we investigated the effects of Dielen® hydrolyzed fish collagens from skin (Promerim®30 and Promerim®60) and cartilage (Promerim®40) to analyze the phenotype and metabolism of equine articular chondrocytes (eACs) cultured as organoids. Here, our findings demonstrated the absence of cytotoxicity and the beneficial effect of Promerim® hydrolysates on eAC metabolic activity under physioxia; further, Promerim®30 also delayed eAC senescence. To assess the effect of Promerim® in a cartilage-like tissue, eACs were cultured as organoids under hypoxia with or without BMP-2 and/or IL-1β. In some instances, alone or in the presence of IL-1β, Promerim®30 and Promerim®40 increased protein synthesis of collagen types I and II, while decreasing transcript levels of proteases involved in OA pathogenesis, namely Htra1, and the metalloproteinases Mmp1-3, Adamts5, and Cox2. Both Promerim® hydrolysates also decreased Htra1 protein amounts, particularly in inflammatory conditions. The effect of Promerim® was enhanced under inflammatory conditions, possibly due to a decrease in the synthesis of inflammation-associated molecules. Finally, Promerim® favored in vitro repair in a scratch wound assay through an increase in cell proliferation or migration. Altogether, these data show that Promerim®30 and 40 hold promise as dietary supplements to relieve OA symptoms in patients and to delay OA progression.
... Similarly, antibacterial peptides obtained from by-products of snow crab and Atlantic rock crab (cancer irroratus) were also showed inhibition against several Gram-positive and Gram-negative bacteria (Beaulieu et al., 2013). Ennaas et al. (2016), isolated and characterized collagencin, an antimicrobial peptide obtained from collagen hydrolysate of fish. The collagencin showed best antibacterial activity at 1.88 mM and inhibited the growth of Staphylococcus aureus (Ennaas et al., 2016). ...
... Ennaas et al. (2016), isolated and characterized collagencin, an antimicrobial peptide obtained from collagen hydrolysate of fish. The collagencin showed best antibacterial activity at 1.88 mM and inhibited the growth of Staphylococcus aureus (Ennaas et al., 2016). Bioactive peptides obtained from skin of black barred halfbeak were also tested for their antibacterial activity against three Gram-positive (Micrococcus luteus (ATCC 4698), Staphylococcus aureus (ATCC 25923), and Bacillus cereus (ATCC 11778)) and three Gram-negative (Salmonella enterica (ATCC 43972), Salmonella typhi (ATCC 19430), (Klebsiella pneumoniae (ATCC 13883)) bacteria (Abdelhedi et al., 2017). ...
Article
Mostly, collagen is obtained from mammalian sources, but its use is limited because of high cost and various allergic reactions. In this review, different alternative sources of collagen were explored and methods for isolation and peptide generations are summarized. Bioactive peptides are short sequences of 2–20 amino acid residues with positive effect on human health. This review summarizes various biological activities of sea food derived peptides/hydrolysates includes antioxidant, inflammatory, antifreeze, angiotension-I converting enzyme (ACE-I) inhibition, antimicrobial, antiaging, wound healing and anticoagulant activities. Moreover, this review also highlights the therapeutic potential and importance of sea-food derived peptides in various pharmaceutical, biomedical, food and cosmetic industries. This review also proposes biological solution for utilization of seafood derived waste in the development of collagen-based food ingredients that is otherwise cause environmental pollution.
... Marine fish skin is a good source of collagen, with antibacterial and antioxidant properties due to collagen peptides and omega-3 polyunsaturated fatty acids, particularly eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Thus, it is unlikely to be a source of infectious disease (3,(5)(6)(7)(8). ...
... This prevents scarring while promoting the healing of wounds. In addition, unlike the gauze bandages, the tilapia skin does not need to be changed every day (3,5,6,(9)(10)(11). ...
Preprint
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Background Tilapia fish skin has demonstrated promise as a stable and practical biological dressing to be used in wound and burn management. However, the appropriate sterilization technique of the Tilapia fish skin is crucial before its clinical application. The standard sterilization technique must eliminate harmful pathogens but maintain the structural and biochemical properties that could compromise the dressing function. This study investigated and compared the efficiency of three sterilizing agents; chlorhexidine gluconate 4% (CHG), povidone iodine 10% (PVP-I), and silver nanoparticles (25 µg/mL) (AgNPs), at three different times (5, 10, and 15 min) on Tilapia fish skin based on the microbial count, histological and collagen properties. Results Among the sterilization procedures, AgNPs showed rapid and complete antimicrobial activity, with a 100% reduction in microbial growth of the fish skin throughout the treated times. Furthermore, AgNPs did not impair the cellular structure or collagen fibers content of the fish skin. However, CHG and PVP-I caused alterations in the collagen content. Conclusion This study demonstrated that the AgNPs treatment of Tilapia fish skin provided sterile skin while preserving the histological properties and structural integrity. These findings provide an efficient and quick sterilization method suitable for Tilapia fish skin that could be adopted as a biological dressing.
... Marine fish skin is a good source of collagen, with antibacterial and antioxidant properties due to collagen peptides and omega-3 polyunsaturated fatty acids, particularly eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Thus, it is unlikely to be a source of infectious disease (3,(5)(6)(7)(8). ...
... This prevents scarring while promoting the healing of wounds. In addition, unlike the gauze bandages, the tilapia skin does not need to be changed every day (3,5,6,(9)(10)(11). ...
Article
Full-text available
Tilapia fish skin has demonstrated promise as a stable and practical biological dressing to be used in wound and burn management. However, the appropriate sterilization technique of the Tilapia fish skin is crucial before its clinical application. The standard sterilization technique must eliminate harmful pathogens but maintain the structural and biochemical properties that could compromise the dressing function. This study investigated and compared the efficiency of three sterilizing agents; chlorhexidine gluconate 4% (CHG), povidone iodine 10% (PVP-I), and silver nanoparticles (25 µg/mL) (AgNPs), at three different times (5, 10, and 15 min) on Tilapia fish skin based on the microbial count, histological and collagen properties. Among the sterilization procedures, AgNPs showed rapid and complete antimicrobial activity, with a 100% reduction in microbial growth of the fish skin throughout the treated times. Furthermore, AgNPs did not impair the cellular structure or collagen fibers content of the fish skin. However, CHG and PVP-I caused alterations in the collagen content. This study demonstrated that the AgNPs treatment of Tilapia fish skin provided sterile skin while preserving the histological properties and structural integrity. These findings provide an efficient and quick sterilization method suitable for Tilapia fish skin that could be adopted as a biological dressing.
... The high bioavailability of collagen peptides makes them available for digestion by the gastric enzymes, get absorbed, and transported. The chemotactic properties of collagen peptides promote cell migration and proliferation, which is important for wound healing (13)(14)(15). Collagen derived from bovine and porcine has been used for a long time, but frequent cases of encephalopathy, hypersensitive reactions, and foot-and-mouth illness have restricted their use in biomedical applications. On the other hand, collagen derived from marine sources imparts desired biomedical property and are currently used for several applications (16). ...
Article
Full-text available
Fabrication of 3D composite scaffolds was carried out by lyophilization of variable concentrations of collagen and chitosan gel solutions. Fibrinogen and thrombin aerosol were deposited over the surface of scaffolds to enhance hemostasis and wound healing. Composite scaffolds were characterized using differential scanning calorimetry, thermogravimetric analysis, and Fourier-transform infrared spectrophotometer to ascertain the aerosol deposition and stability. Scanning electron microscope showed multilayered porosity with pore size of ~30 μm and mushroom-like fibril growth of aerosol. A detailed investigation by surface plasmon resonance confirmed higher binding affinity of collagen toward the human blood platelets and erythrocytes in comparison to chitosan and was found to increase with the increase in blood cell concentration from 480.8 to 886.4 RU for erythrocytes. Scaffolds showed higher binding response for platelets than erythrocytes, while fibrinogen and thrombin showed no or limited interaction. Highest blood sorption of 83 ± 4% was observed in case of aerosol deposited scaffolds. Aerosol deposited scaffolds showed minimum clotting time of 20 ± 3 s and bleeding time of 38 ± 4 s, which was significantly lower compared to the scaffolds without aerosol treatment. Aerosol deposited composite scaffolds with 2:1 concentration of chitosan/collagen showed complete wound contraction by day 14, while 50% was observed in case of the control group. In vivo studies revealed that chitosan had a crucial role in the inflammatory phase, while collagen played an important role in the proliferation and maturation phase. The present study suggests that the fabricated 3D composite scaffolds with bioactive moieties may be a potential candidate for enhanced hemostasis and wound healing applications.
... The mechanism of collagen peptide follows the carpet model. 33 Peptides accumulate on the surface of the membrane to reach threshold concentrations to cover the surface forming the carpet. This interaction will affect the integrity of the membrane, resulting in cell lysis.. 34 ...
... [10][11][12] In spite of implementing this method to extract collagen, many scientists before only focused on reporting the relative molecular weight and amino acid compositions. [13][14][15][16] Nevertheless, the characteristics of collagen extracted from a mixture of fresh-water fish scales by biochemical method have been limited in publication. In our previous manuscript, we reported the characteristics of collagen extracted from carp fresh water fish scales by chemical method. ...
Article
Full-text available
This paper describes the characteristics including morphology, amino acid composition, element composition, and denaturation temperature of collagen extracted from freshwater fish scales caught in Vinh Phuc province, Vietnam by biochemical method in acetic acid solution containing pepsin. The pepsin concentration used for the extraction process of collagen varied from 1 to 3 wt.% (correspondence to the weight of initial fish scale). The weight and denatured temperature of collagen at different content of pepsin were determined. The obtained results showed that the ratio of 2 wt.% of pepsin is the most suitable concentration for the extraction of collagen from freshwater fish scales. Some typical properties of collagen such as morphology, functional groups, amino acid composition, and elemental composition were investigated by Infrared spectroscopy (IR), Ultraviolet-visible spectroscopy (UV-Vis), Energy-dispersive X-ray spectroscopy (EDX), High performance liquid chromatography (HPLC), and Field emission scanning electron microscopy (FESEM) methods and discussed.
... Hence, the incorporation of an antibacterial effect in Col further contributes to its physicochemical characteristics. Col can be integrated with silver nanoparticles (AgNP) [63], collagencin [64], epoxidized safrole [65], polyhexamethylene biguanide [66], titanium dioxide [67], etc. Similarly, native gelatin does not show any antibacterial effect. ...
Article
Full-text available
Collagen (Col) and gelatin are most extensively used in various fields, particularly in pharmaceuticals and therapeutics. Numerous researchers have proven that they are highly biocompatible to human tissues, exhibit low antigenicity and are easy to degrade. Despite their different sources both Col and gelatin have almost the same effects when it comes to wound healing mechanisms. Considering this, the bioactivity and biological effects of both Col and gelatin have been, and are being, constantly investigated through in vitro and in vivo assays to obtain maximum outcomes in the future. With regard to their proven nutritional values as sources of protein, Col and gelatin products exert various possible biological activities on cells in the extracellular matrix (ECM). In addition, a vast number of novel Col and gelatin applications have been discovered. This review compared Col and gelatin in terms of their structures, sources of derivatives, physicochemical properties, results of in vitro and in vivo studies, their roles in wound healing and the current challenges in wound healing. Thus, this review provides the current insights and the latest discoveries on both Col and gelatin in their wound healing mechanisms.
... Antimicrobial peptide P4 showed complete inhibition of Listeria innocua and Escherichia coli, implicating the action of the amino acid sequence. The antimicrobial action (complete inhibition) of collagencin peptide extracted from fish collage against Staphylococcus aureus was reported by [111]. Partial action against Staphylococcus pyogenes, Escherichia coli, Listeria innocua, Lactococcus lactis and Carnobacterium divergens was reported. ...
Article
Full-text available
Developing peptide-based drugs are very promising to address many of the lifestyle mediated diseases which are prevalent in a major portion of the global population. As an alternative to synthetic peptide-based drugs, derived peptides from natural sources have gained a greater attention in the last two decades. Aquatic organisms including plants, fish and shellfish are known as a rich reservoir of parent protein molecules which can offer novel sequences of amino acids in pep-tides, having unique bio-functional properties upon hydrolyzing with proteases from different sources. However, rather than exploiting fish and shellfish stocks which are already under pressure due to overexploitation, the processing discards, regarded as secondary raw material, could be a potential choice for peptide based therapeutic development strategies. In this connection, we have attempted to review the scientific reports in this area of research that deal with some of the well-established bioactive properties, such as antihypertensive, anti-oxidative, anti-coagulative, antibac-terial and anticarcinogenic properties, with reference to the type of enzymes, substrate used, degree of particular bio-functionality, mechanism, and wherever possible, the active amino acid sequences in peptides. Many of the studies have been conducted on hydrolysate (crude mixture of peptides) enriched with low molecular bioactive peptides. In vitro and in vivo experiments on the potency of bioactive peptides to modulate the human physiological functions beneficially have demonstrated that these peptides can be used in the prevention and treatment of non-communicable lifestyle mediated diseases. The information synthesized under this review could serve as a point of reference to drive further research on and development of functionally active therapeutic natural peptides. Availability of such scientific information is expected to open up new zones of investigation for adding value to underutilized secondary raw materials, which in turn paves the way for sustainability in fish processing. However, there are significant challenges ahead in exploring the fish waste as a source of bioactive peptides, as it demands more studies on mechanisms and structure-function relationship understanding as well as clearance from regulatory and statutory bodies before reaching the end user in the form of supplement or therapeutics. Citation: Phadke, G.G.; Rathod, N.B.; Ozogul, F.; Elavarasan, K.; Karthikeyan, M.; Shin, K.-H.; Kim, S.-K. Exploiting of Secondary Raw Materials from Fish Processing Industry as a
... Clavanins, hedistin, piscidin, myxinidin, pleurocidin and styelins are marine AMPs included in this group (Lehrer et al., 2001;Pundir et al., 2014). Proline-and arginine-rich callinectin (Khoo et al., 1999;Noga et al., 2011), histidinerich chrysophsin (Iijima et al., 2003;Mason et al., 2007), and proline-and glycine-rich collagencin (Ennaas et al., 2016) are linear or helical peptides with an abundance of one amino acid (II). The third group is peptides forming a hairpin-like βsheet or αhelical/β-sheet mixed structures stabilized by intramolecular disulphide bonding (III). ...
Article
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Antimicrobial Peptides (AMPs) are compounds, which have inhibitory activity against microorganisms. In the last decades, AMPs have become powerful alternative agents that have met the need for novel anti-infectives to overcome increasing antibiotic resistance problems. Moreover, recent epidemics and pandemics are increasing the popularity of AMPs, due to the urgent necessity for effective antimicrobial agents in combating the new emergence of microbial diseases. AMPs inhibit a wide range of microorganisms through diverse and special mechanisms by targeting mainly cell membranes or specific intracellular components. In addition to extraction from natural sources, AMPs are produced in various hosts using recombinant methods. More recently, the synthetic analogs of AMPs, designed with some modifications, are predicted to overcome the limitations of stability, toxicity, and activity associated with natural AMPs. AMPs have potential applications as antimicrobial agents in food, agriculture, environment, animal husbandry, and pharmaceutical industries. In this review, we have provided an overview of the structure, classification, and mechanism of action of AMPs, as well as discussed opportunities for their current and potential applications.
... Esta proteína se obtiene de diferentes fuentes naturales como son: de origen porcino (cerdos), de origen bovino (vacas) y de origen marino (peces) (Gómez et al., 2011). La industria productora de colágeno tiene tendencia a sustituir los agentes sintéticos con otros naturales, para ello, se están buscando nuevas fuentes naturales de donde extraer el colágeno, considerándose a los subproductos de especies acuáticas una fuente prometedora de este material, y que se pueden convertir en una alternativa rentable debido a que son usualmente descartados (Ennas et al., 2016). De acuerdo a la información emitida en La Organización de las Naciones Unidas para la Alimentación y la Agricultura -FAO (2018), indica que los recursos pesqueros se han convertido en uno de los más importantes en la economía de varios países alrededor del mundo. ...
Article
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En los últimos años, el colágeno ha tomado importancia en nuestra dieta alimenticia por mantener firmeza y flexibilidad en nuestra piel, uñas y cabello. En el presente trabajo, se obtuvo colágeno natural a partir de escamas de pescado de “cachema” (Cynoscion analis), “bonito” (Sarda chiliensis) y “corvina” (Cilus gilberti). Las especies fueron seleccionadas por ser las más vendidas en los mercados populares de la ciudad, se compraron y se lavaron para luego extraer las escamas con ayuda de un cuchillo dentado, las escamas se secaron a temperatura ambiente y se esterilizaron a 80°C por 30 minutos en vapor seco. Para la obtención del colágeno se hidrataron las escamas y luego se pulverizó por método mecánico, que consistió en colocar las escamas en una olla adicionando 1 litro de agua, luego se hirvieron por espacio de media hora, finalmente se licuó y tamizó. El producto obtenido se conservó a 4°C, y fue sometido a análisis de caracteres sensoriales y contenido de proteína total por método de Kjeldahl. Se concluye que no hay diferencia de sabor, textura y aroma, entre el colágeno obtenido en los tres tratamientos, siendo el contenido de proteínas mayor en corvina (67%).
... [19][20][21][22][23][24][25] Collagen is a kind of macromolecular protein found in skin, blood vessels and bones, which obtained from different animals has good health function. 26,27 Chitosan have been identified as having universal antibacterial property, good coagulation ability and biocompatibility functional agent, it also has anticancer, anti-ulcer and anti-infection effects. 28,29 Sericin and its hydrolysates have good pharmacological effects, such as antibacterial and anti-inflammatory, anticoagulant, anticancer, etc. 30,31 Immobilization of biological molecules onto PP fabric has already made some progress. ...
Article
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In order to realize the immobilization of collagen, chitosan and sericin on the surface of polypropylene fabric, the fabric is treated by ammonia and nitrogen low temperature plasma to produce reactive groups, and then the epoxy compounds (ethylene glycol diglycidyl ether, trimethylolpropane triglycidyl ether) are used as crosslinkers for a bridging process. Single factor experiment is carried out to investigate the influence of crosslinking parameters (temperature, time and the dosage of crosslinker) on immobilization yield. Chemical composition changes on the surface of the treated samples is analyzed by infrared spectroscopy, which indicated that the ammonia plasma treatment can produce more active groups. The results showed that immobilization rate of samples crosslinked by ethylene glycol diglycidyl ether is superior to that of trimethylolpropane triglycidyl ether. It also can be found that the fabric had a better wettability and antimicrobial properties. The whiteness and comfort property of chitosan-polypropylene fabric decreased slightly.
... At the same time, collagen derived peptides are an abundant source of functional chemicals. Collagen peptides or hydrolysates have been proven to have lots of special functions, such as benefits for skin and bone (Zdzieblik et al., 2017), antioxidant (Ketnawa et al., 2016), antimicrobial (Ennaas et al., 2016), antifreeze (Cao et al., 2016). These special functions of collagen peptides mean that collagen peptides can be used as functional foods, medicine, cosmetics, food additives and so on. ...
... The negative GRAVY value of analog 4 means that it is more hydrophilic. Analog 5 has the highest GRAVY value, which is due to the substitution of the hydrophilic E 26 and D 32 residues with hydrophobic A residues. Analog 7 has the second-highest GRAVY value amongst the full-length (32 amino acid residues) analogs because of the three A 18 , A 19 and A 29 that have been substituted with more hydrophobic leucine (L) residues. ...
Article
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The structure–activity relationships and mode of action of synthesized glyceraldehyde-3-phosphate dehydrogenase (GAPDH)-related antimicrobial peptides were investigated. Including the native skipjack tuna GAPDH-related peptide (SJGAP) of 32 amino acid residues (model for the study), 8 different peptide analogs were designed and synthesized to study the impact of net charge, hydrophobicity, amphipathicity, and secondary structure on both antibacterial and antifungal activities. A net positive charge increase, by the substitution of anionic residues or C-terminal amidation, improved the antimicrobial activity of the SJGAP analogs (minimal inhibitory concentrations of 16–64 μg/mL), whereas the alpha helix content, as determined by circular dichroism, did not have a very definite impact. The hydrophobicity of the peptides was also found to be important, especially for the improvement of antifungal activity. Membrane permeabilization assays showed that the active peptides induced significant cytoplasmic membrane permeabilization in the bacteria and yeast tested, but that this permeabilization did not cause leakage of 260 nm-absorbing intracellular material. This points to a mixed mode of action involving both membrane pore formation and targeting of intracellular components. This study is the first to highlight the links between the physicochemical properties, secondary structure, antimicrobial activity, and mechanism of action of antimicrobial peptides from scombrids or homologous to GAPDH.
... Haemolytic activity of collagen type 1 was estimated by measuring the release of haemoglobin from human RBC (Ennaas et al. 2016). 5 ml of blood was diluted 40 times with 0.05 M phosphate buffer saline and incubated for 1 h with 10 mg/ml of collagen type 1. ...
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In this study, a comparative structural and bioactive analysis of collagen extracted from the skin of bony and cartilaginous fishes. The acid-soluble method was followed to extract the collagen from Hypophthalmichthys molitrix (silver carp) and Rhizoprionodon acutus (milk shark) followed by purification using Ion exchange chromatography. A higher yield of collagen was obtained from the skin of SCsk (69.45%) as compared to SHsk (55.29%). SDS PAGE displayed the characteristic α, β bands of the collagen type1. The native conformation and secondary structure stability of collagen were confirmed by FTIR, XRD and CD studies. The SEM micrographs exhibited the layered and fibrillar nature of the collagen from SHsk and SCsk, respectively. Relative solubility and thermal denaturation analysis showed SCsk to be more stable, but SHsk could withstand higher temperatures. 53.65% of antioxidant activity was observed in SCsk collagen as compared to SHsk (45.9%). Haemocompatibility, cell viability and adhesion results also displayed silvercarp skin to be a better source than Shark skin collagen. The results establish the potential of silver carp collagen as a biomaterial that can have many commercial applications in tissue engineering, cosmetics and food industries.
... The identification of these peptides can be performed using spectroscopic techniques, such as CD and FTIR. The FTIR to identify a collagen peptide with an immunologic role and good cellular apoptosis tolerance extracted from cartilage of Prionace glauca (Blue shark) [17] ; while Ennaas et al. [216] used CD to identify and characterize a peptide (Collagencin) with antibacterial (Gram + and Gram-) properties, preventing the growth of Staphylococcus aureus . Some collagen-based food products are described in Table 5 . ...
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Collagenous biopolymers can be analyzed using physic-chemical, densitometric, and spectroscopic analysis to obtain their main characteristics, aiming at their biotechnological manipulation. Collagen extracted from fishery resources is a product of high added value, with potential use in food, biopharmaceutical and cosmetic industries. Some factors make the use of the polymer from fishery resources promising, such as: high availability; greater ontogenetic distance between fish and humans; absence of sociocultural barriers; and absence of toxicity. The collagen extraction method (acid-soluble, pepsin-soluble, electrodialysis, ultrasound, isoelectric precipitation) will directly influence its properties. Thus, this work aims to provide an overview of the extraction methods, characterization techniques (solubility, zeta potential, viscosity, thermogravimetry, differential scanning calorimetry, SDS-PAGE, densitometry, gel strength, centesimal composition, color, aminogram, hydroxyproline determination, X-ray diffraction, circular dichroism, ultraviolet, Raman, and FTIR spectroscopy), and potential analysis with a focus on aquaculture and fisheries sources, through a compilation of scientific information that can be useful to guide aquatic biotechnology professionals, considered that its properties are similar to collagen extracted from mammals.
... In model systems, 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC) liposomes are often used to represent mammalian membrane. Ennaas et al. studied the interaction dynamics of antibacterial peptide collagencin and membrane using DPPC liposome (Ennaas et al., 2016). Investigating the interaction between other antimicrobial peptides or antigenic peptides with membrane based on DPPC liposome has also reported in previous studies (Dayane, Maria, João & Natalia, 2016;Sospedra, Mestres, Haro, Muñoz, & Busquets, 2002). ...
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Understanding the interaction of food derived angiotensin converting enzyme (ACE) inhibitory peptides and intestinal epithelial cell membrane may help to improve their absorption. This research aimed to study the molecular interaction of ACE inhibitory tri-peptides ADF, FGR, and MIR with DPPC membrane during absorption process. The DPPC liposome was prepared and characterized, then used as a model membrane. The permeability of tri-peptides across the membrane was investigated using Fluorescence spectroscopy. The effect of tri-peptides on the structure and dynamics of DPPC bilayers was determined using Fourier transform infrared spectroscopy. The effect of tri-peptides on the phase transition temperature in the DPPC membrane was also analyzed using Differential scanning calorimetry. The results showed that ACE inhibitory tri-peptides ADF, FGR, and MIR can penetrate into both the membrane-water interface and hydrophobic region of DPPC bilayer, and the tri-peptide FGR have higher permeability across the membrane.
... Finally, MC represents a useful source of various antimicrobial peptides [179]. Among them, collagencin has been shown to inhibit the growth of a broad spectrum of bacteria, by interacting with both anionic (phosphatidylglycerol) and zwitterionic (phosphoethanolamine and phosphatidylcholine) lipids [180]. ...
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... Since then, many different AMPs have been found in various fish species in different environments. For instance, AMPs have been isolated from salmon liver (Richards et al., 2001), Atlantic cod mucus (Bergsson et al., 2005), tilapia side streams (Robert et al., 2015), Atlantic mackerel and its side streams (Ennaas et al., 2016), and from rainbow trout side streams (Wald et al., 2016). In addition to AMPs naturally found in various parts of fish, it is also possible to produce AMPs enzymatically from different fish species and their processing by-products (Table 2). ...
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The current practice of fish processing generates increasing quantities of side streams and waste, such as skin, heads, frames, viscera, and fillet cut offs. These may account for up to 70% of the fish used in industrial processing. Low-value fish catches, and under-utilized fish species comprise another source of side streams. These side streams have been discarded in the environment leading to environmental problems or they have ended up as low commercial value products, such as feed for fur animals and aquaculture. However, several studies have shown that fish side streams contain valuable bioactive ingredients and fractions, such as fish oils, proteins and peptides, collagen, gelatin, enzymes, chitin, and minerals. These compounds and fractions may provide the opportunity to develop novel applications in health promoting foods, special feeds, nutraceuticals, pharmaceuticals, and cosmetic products. Better utilization of side streams and low-value fish would simultaneously improve both the environmental and ecological sustainability of production. This review summarizes the current knowledge on fish and fish side streams as sources of high-value components such as peptides with antimicrobial, antioxidative, antihypertensive, and antihyperglycemic properties, proteins such as fish collagen and gelatin, fish enzymes, fish oils and fatty acids, polysaccharides like glucosaminoglycans, chitin and chitosan, vitamin D, and minerals. Production technologies for recovering the high-value fractions and potential product applications are discussed. Furthermore, safety aspects related to the raw material, technologies, and fractions are considered.
... The first step in interaction of numerous natural and artificial hydrophobic or amphiphilic compounds with the cell is their interaction with the lipid matrix of plasma membrane followed by changing its structural and functional characteristics which called the membranotropic effect [67]. Affecting the lipid layer or its components, bioactive compound is capable to modify the membrane-associated proteins' activity and, thus, to implement its effect on the cell by receptor-free mechanism [68,69]. ...
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Pyrrole derivatives (PDs) chloro-1-(4-chlorobenzyl)-4-((3-(trifluoromethyl)phenyl)amino)-1H-pyrrole-2,5-dione (MI-1) and 5-amino-4-(1,3-benzothyazol-2-yn)-1-(3-methoxyphenyl)-1,2-dihydro-3H-pyrrole-3-one (D1) were synthesized as inhibitors of several protein kinases including EGFR and VEGFR. The aim of the study was to reveal the exact mechanisms of PDs’ action EGFR and VEGFR are involved in. We observed, that both PDs could bind with EGFR and VEGFR and form stable complexes. PDs entered into electrostatic interactions with polar groups of phospholipid heads in cell membrane, and the power of interaction depended on the nature of PD radical substituents (greater for MI-1 and smaller for D1). Partial intercalation of MI-1 into the membrane hydrophobic zone also occurred. PDs concentrations induced apoptosis in malignant cells but normal ones had different sensitivity to those. MI-1 and D1 acted like antioxidants in inflamed colonic tissue, as evidenced by reduce of lipid and protein peroxidation products (by 43-67%) and increase of superoxide dismutase activity (by 40 and 58%) with restoring these values to control ones. MI-1 restored reduced hemoglobin and normalized elevated platelets and monocytes in settings of colorectal cancer, whereas D1 normalized only platelets. Thus, MI-1 and D1 could be used as competitive inhibitors of EGFR and VEGFR and antioxidants, which might contribute to realization of their anti-inflammatory, proapoptotic and antitumor activity.
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Background: Collagen is the most abundant structural protein in the mammalian connective tissue and represents approximately 30% of animal protein. The current study evaluated the potential capacity of collagen extract derived from Nile tilapia skin in improving the cutaneous wound healing in rats and investigated the underlying possible mechanisms. A rat model was used, and the experimental design included a control group (CG) and the tilapia collagen treated group (TCG). Full-thickness wounds were conducted on the back of all the rats under general anesthesia, then the tilapia collagen extract was applied topically on the wound area of TCG. Wound areas of the two experimental groups were measured on days 0, 3, 6, 9, 12, and 15 post-wounding. The stages of the wound granulation tissues were detected by histopathologic examination and the expression of vascular endothelial growth factor (VEGF), and transforming growth factor (TGF-ß1) were investigated using immunohistochemistry. Moreover, relative gene expression analysis of transforming growth factor-beta (TGF-ß1), basic fibroblast growth factor (bFGF), and alpha-smooth muscle actin (α-SMA) were quantified by real-time qPCR. Results: The histopathological assessment showed noticeable signs of skin healing in TCG compared to CG. Immunohistochemistry results revealed remarkable enhancement in the expression levels of VEGF and TGF-β1 in TCG. Furthermore, TCG exhibited marked upregulation in the VEGF, bFGF, and α-SMA genes expression. These findings suggested that the topical application of Nile tilapia collagen extract can promote the cutaneous wound healing process in rats, which could be attributed to its stimulating effect on recruiting and activating macrophages to produce chemotactic growth factors, fibroblast proliferation, and angiogenesis. Conclusions: The collagen extract could, therefore, be a potential biomaterial for cutaneous wound healing therapeutics.
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The effects of L. barbarum (LB) and/or fish collagen (FC) in cheese on the proteolytic degradation profile with anti‐ACE activity were investigated. Four types of cheeses were made plain cheese (control), LB‐cheese, FC‐cheese (control), and LB‐cheese+FC. Acidification of cheese, milk proteins proteolysis, and angiotensin‐converting enzyme (ACE) inhibitory activity were performed during 28 days. The OPA peptides content of LB‐cheese was higher (p<0.05) than control on 28 days. The addition of FC in LB‐cheese improved OPA peptides content. The proteins degradation of β‐lactoglobulin, α‐ and β‐caseins increased in the presence of LB in cheese whereas α‐ and β‐caseins degradation improved in LB + FC cheese. ACE inhibitory activity was decreased (p<0.05) in LB‐cheese but not in LB+ FC. The combination of LB and FC in cheese showed the most enhancements on the production of peptides that may act as anti‐ACE activity.
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Background The health benefits associated with consuming fish products are mainly attributed to their desirable nutrition profiles, including vitamins, minerals, essential amino acids, and polyunsaturated fatty acids. However, large quantities of fish proteins are presently underutilized or discarded as waste. Effective strategies to utilize fish proteins are therefore needed. Recently, researchers have focused on generating and characterizing bioactive fish protein hydrolysates and peptides and then studying their potential health benefits. Scope and Approach The major methods of producing, separating, and purifying protein hydrolysates are initially given. Then, the biological activities and potential mechanisms of action of protein hydrolysates and peptides are discussed. Finally, current limitations and future possibilities of fish peptide identification, production, and bioactivity are identified and discussed. Key findings and conclusions Fermentation, chemical synthesis, and enzymatic hydrolysis are effective methods of obtaining hydrolysates from underutilized fish protein by-products. These hydrolysates can then be purified by membrane separation and chromatographic methods to obtain bioactive peptides. The molecular characteristics of the peptides can then be identified using mass spectrometry. Fish hydrolysates/peptides have multiple biological activities, including antioxidative, lipid homeostasis modulation, anti-inflammatory, anticancer, neuroprotective, and antihypertensive activities, which make them promising nutraceutical ingredients for application in foods. Moreover, they often have emulsifying, foaming, and gelling properties, which means they may be suitable as multipurpose functional ingredients. Thus, waste-derived fish by-products may be turned into value-added functional ingredients designed to address chronic diseases. However, further research is required to develop large-scale commercially viable extraction and purification methods, develop robust structure-function relationships for peptides, and perform in vivo human studies of peptide bioactivity.
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Four types of cheeses were prepared included plain- cheese (control), Codonopsis pilosula (CP)- cheese, plain- cheese with fish collagen (FC; control) and CP- cheese with FC. The effects of cheese samples on acidification, proteolysis of milk proteins using three methods (cadmium-ninhydrin method, O-phthaldialdehyde (OPA) assay, and electrophoresis assay), and angiotensin-converting enzyme (ACE)-inhibitory activity were investigated during 0, 2, & 4 weeks of ripening. In addition, the sensory evaluation was also investigated during 0, 2, 4, & 8 weeks of ripening. The presence of FC in CP- cheese increased the numbers of free amino acids (FAA) at 0 and 2 weeks. The addition of CP both in the presence and absence of FC affected positively (p<0.05) on the concentrations of OPA peptide in cheese compared to their respective controls. The presence of CP and/or FC in cheese increased the degradation of milk proteins (α-, β-, & κ- caseins, β-lactoglobulin, and α-lactalbumin) compared to their respective controls during ripening periods. The highest ACE inhibitory activity was shown at 4 weeks of ripening for CP- cheese both in the absence (67.75 ± 14.15 %) and the presence (78.65 ± 2.85 %) of FC. In addition, 8-week-old CP-cheese in the presence and absence of fish collagen had similar organoleptic characteristics to plain-cheese. In conclusion, C. pilosula and/or fish collagen may lead to the development in the production and formulation of cheese with anti-ACE activity.
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Fish gelatin is a class of biopolymers acquired from the hydrolysis of fish collagen, and it contains abundant amino acids for nutritive uses as dietary products. The natural characteristics of fish gelatin mean that fish gelatin products are relatively harmless to the body compared with clinical therapies and drugs. Thus, the dietary use of fish gelatin can potentially have excellent benefits for people with chronic diseases such as hypertension, osteoporosis, and diabetes. Moreover, researchers have found multiple routes for the administration of fish gelatin products such as external use or injection. In this review, we summarize the new usage of fish gelatin products and provide an update on the latest research on other properties and applications of fish gelatin. These new applications will provide multiple uses for fish gelatin products in the future, though further research is still needed.
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This study investigated peptide fractions from fish skin collagen for antibacterial activity against Escherichia coli and Salmonella strains. The collagen was hydrolyzed with six commercial proteases, including trypsin, Alcalase, Neutrase, Flavourzyme, pepsin and papain. Hydrolyzed samples obtained with trypsin and Alcalase had the largest number of small peptides (molecular weight < 10 kDa), while the hydrolysate produced with papain showed the lowest degree of hydrolysis and highest number of large peptides. Four hydrolysates were found to inhibit the growth of the Gram-negative bacteria, with papain hydrolysate showing the best activity against E. coli, and Neutrase and papain hydrolysates showing the best activity against S. abony; hydrolysates produced with trypsin and pepsin did not show detectable antibacterial activity. After acetone fractionation of the latter hydrolysates, the peptide fractions demonstrated enhanced dose-dependent inhibition of the growth (colony-forming units) of four Salmonella strains, including S. abony (NCTC 6017), S. typhimurium (ATCC 13311), S. typhimurium (ATCC 14028) and S. chol (ATCC 10708). Shotgun peptidomics analysis of the acetone fractions of Neutrase and papain hydrolysates resulted in the identification of 71 and 103 peptides, respectively, with chain lengths of 6–22 and 6–24, respectively. This work provided an array of peptide sequences from fish skin collagen for pharmacophore identification, structure–activity relationship studies, and further investigation as food-based antibacterial agents against pathogenic microorganisms.
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There are several reviews that separately cover different aspects of fish gelatin including its preparation, characteristics, modifications, and applications. Its packaging application in food industry is extensively covered but other applications are not covered or covered alongside with those of collagen. This review is comprehensive, specific to fish gelatin/hydrolysate and cites recent research. It covers cosmetic applications, intrinsic activities, and biomedical applications in wound dressing and wound healing, gene therapy, tissue engineering, implants, and bone substitutes. It also covers its pharmaceutical applications including manufacturing of capsules, coating of microparticles/oils, coating of tablets, stabilization of emulsions and drug delivery (microspheres, nanospheres, scaffolds, microneedles, and hydrogels). The main outcomes are that fish gelatin is immunologically safe, protects from the possibility of transmission of bovine spongiform encephalopathy and foot and mouth diseases, has an economic and environmental benefits, and may be suitable for those that practice religious-based food restrictions, i.e., people of Muslim, Jewish and Hindu faiths. It has unique rheological properties, making it more suitable for certain applications than mammalian gelatins. It can be easily modified to enhance its mechanical properties. However, extensive research is still needed to characterize gelatin hydrolysates, elucidate the Structure Activity Relationship (SAR), and formulate them into dosage forms. Additionally, expansion into cosmetic applications and drug delivery is needed.
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Collagen is the most abundant extracellular matrix protein in food-producing animals. Gelatin is partially degraded collagen. Collagen peptides refer to the peptides with specific properties identified from collagen hydrolysate who produced by hydrolysis of collagen/gelatin. Due to the specific structural and bio- and physical-chemical properties, collagen and its derivatives are used in the field of food industry. In this review, the structure of the collagen molecule and its biosynthetic process in vivo are introduced, and the production methods and structures of gelatin and collagen peptides described. Then the inherent self-assembly property of collagen, the mechanical properties of collagen and gelatin gels, functional properties of collagen and gelatin, and bioactive properties of collagen peptides are reviewed. Finally, the applications of collagen and its derivatives that are correlated with their properties in food industry are summarized. The mechanisms and advantages of the applications of collagen and its derivatives in food industry are raised, and the limitations and challenges of these applications are also discussed. And possible studies to address the challenges of the applications in different areas are indicated.
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                                             
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Recently, proteins are gaining attention as potential materials for antibacterial therapy. Proteins possess beneficial properties such as biocompatibility, biodegradability, low immunogenic response, ability to control drug release, and can act as protein-mimics in wound healing. Different plant- and animal-derived proteins can be developed into formulations (films, hydrogels, scaffolds, mats) for topical antibacterial therapy. The application areas for topical antibacterial therapy can be wide including bacterial infections in the skin (e.g., acne, wounds), eyelids, mouth, lips, etc. One of the major challenges of the healthcare system is chronic wound infections. Conventional treatment strategies for topical antibacterial therapy of infected wounds are inadequate, and the development of newer and optimized formulations is warranted. Therefore, this review focuses on recent advances in protein-based systems for topical antibacterial therapy in infected wounds. The opportunities and challenges of such protein-based systems along with their future prospects are discussed.
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In this study, we introduced a novel polymerization method of polyester using collagen peptides derived from fish scale waste. After the extraction process of collagen peptide from fish scales, putting collagen peptide, ethylene glycol and Benzenedicarboxylic acid into a container, and mixing them to form a mixture; heating the mixture for executing an esterification reaction, to product esters and water; heating the esters, and stirring the esters via a mixer; in a specific period, decreasing the pressure in the container for executing a polycondensation reaction; decreasing the pressure in the container to a second pressure, and stirring the esters via the mixer, to produce a collagen modified polyester. Collagen peptides are rich in glycine, proline, and hydroxyproline, and by forming a triple helix structure, such as that of the copolyester, gain better hydrophilicity, antistaticity, and ductility. As a result, the produced collagen modified polyester fiber keeps the characteristics of the traditional polyethylene terephthalate fibers including strength, durability, and resistance to wrinkle and shrink. However, the supramolecular collagen modified polyester containing animal collagen peptides has naturally a soft touch and champagne-like color. Consequently, it can be used as a suitable material for skin-friendly functional clothes with or without additional dying. In brief, • This study introduces a novel method for collagen modified polyester. • Upcycled fish scale waste brings the sustainable benefits of circular economy. • Collagen modified polyester provides a new direction for future technological development in the textile industry.
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In the present scenario, there is a great demand for targeted treatment to cure various diseases; researchers are searching for an efficient drug to treat pathologies. Aquatic peptides identified from various bio-resources of the aquatic environment such as aquatic plants, microbes and animals are one of the important therapeutic molecules. The aquatic organisms are native to innate immune system that exclusively synthesis numerous immune protein and bioactive peptides to protect the organisms from a variety of life-threatening environmental factors and subsequent pathogenic attack. Aquatic peptides from both freshwater and marine aquatic ecosystems have promising active molecules against pathogenic infection. Each peptide has unique properties to treat the disease. Meanwhile, peptide-based drug development is not an easy task; it requires overcoming various hurdles even in the current advanced techniques available today. This review discusses the potential of aquatic peptides and the limitations in developing them as drug and the current techniques and strategies to achieve a successful peptide-based drug development.
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By-product removal in fish processing is estimated to be between 25 and 70% due to improper fish production handling and significant problems in the fish industry today. Therefore, one of the ways to manage the raw material of by-product is through protein hydrolysis. However, one of the most effective methods for managing this raw material, which includes skin, bones, heads, and viscera, is to convert their protein into peptides via hydrolysis methods, resulting in fish protein hydrolysate (FPH). FPH has been shown to have bioactive properties such as antibacterial, antihypertensive, antioxidative, anticancer, and anticoagulant properties. Bioactivity could be fully utilised in the future in both the nutraceutical and food industries. Numerous studies have been published on the acceptability of FPH in obtaining bioactive properties from various fish, particularly antibacterial activity. For example, the antibacterial peptide was identified as FPIGMGHGSRPA, consisting of 12 amino acids. Its antibacterial activity was tested against B. subtilis using 800 g/mL ampicillin. The inhibition zone increased with peptide concentration. This review discusses functional bioactive peptides derived from fish protein hydrolysate that can be used as antibacterial agents by inhibit Gram-positive and Gram-negative bacterial growth. It also covers fish species, parts, and hydrolysis methods to maximise yields.
Article
PurposeStaphylococcus epidermidis is one of the most common causative pathogens of nosocomial-related infections. It is known to form biofilms on indwelling medical devices. The negatively charged extracellular matrix of the biofilm can entrap bacteria, thus limiting the diffusion of therapeutic agents and contributing to the ineffectiveness of therapeutic agents against the bacteria. To combat this, we investigated the effectiveness of a cationic peptide, K16ApoE, with bioadhesive properties to serve as an anti-adhesion agent.Methods The ability of K16ApoE to serve as an anti-adhesive agent was evaluated using crystal violet staining to quantify the degree of inhibition of biofilm formation. The minimum inhibitory concentration was determined by concurrently incubating the S. epidermidis inoculum with K16ApoE (0–250 µg/mL) for 24 h, after which a relative biofilm density assay was performed. Inhibition of the surface adhesion of biofilms to various matrices was also evaluated by coating K16ApoE on ceramic discs as well as polyvinyl chloride (PVC) or silicone catheter extension tubing. Bovine serum albumin (BSA), pure unconjugated ApoE or K16 peptide were used as controls.ResultsK16ApoE (250 µg/mL) was very effective at reducing biofilm integrity by 99.92%. ApoE or K16 alone was largely ineffective compared to K16ApoE. In addition, catheter tubing pretreated with K16ApoE showed a significant (83 ± 12.7%) reduction in the formation of adherent biofilms.Conclusion These results demonstrate that K16ApoE can serve as an effective anti-adhesive agent to prevent the formation of an adherent bacterial biofilm on matrices with different surface properties.
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Background Bioactive peptides have been studied in several sources due to their valuable potential in pharmaceutical, food and cosmetic industries. Algae, mussels and fish are examples of marine sources of bioactive peptides. Several biological properties have been identified in these peptides, such as antioxidant, antiatherosclerosis, anticancer, anticoagulant, anti-inflammatory, antihypertensive and antimicrobial activities. Furthermore, peptides derived from marine sources may also be of great interest due to their functional properties, such as solubility, emulsifying and foaming properties, which could be beneficial for industrial application. Scope and approach This paper aims to review information about bioactive peptides isolated from different marine sources, focusing on the different biological activities and functional properties already described, and the different industrial applications exploited. Key findings and conclusions Enzymatic hydrolysis appears as the most used method for peptide production. Bioactive peptides have not been widely used individually, although marine protein hydrolysates are already used with different industrial purposes. However, due to all the bioactivities showed, marine peptides may be of great interest to industrial applications, such as active ingredients for food or cosmetic products; preservatives for food or cosmetics; anti-inflammatory cosmeceuticals; pharmaceutical or nutraceutical products to treat or prevent diseases, with potentially fewer side effects when compared with synthetic drugs. To incorporate peptides in industrial matrices it is essential to guarantee that they will not suffer modifications or lose bioactivity inside new matrices. To empower peptides action in pharmaceuticals, food or cosmetics, they must be able to resist to adverse external factors. Encapsulation may be of one of the most interesting approaches to increase peptides protection.
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Fish farming (aquaculture) provides a stable source of nutritional protein, especially in Asia's coastline, distributed throughout the South China region of the mainland, southwestern Taiwan, and Southeast Asia, providing a continuous supply chain system. Novel textile material, from upcycling of fish scales, has been manufactured. The material constitutes of collagen peptide amino acid ingredients extracted from fish scales, which are used to create collagen modification polyester via a supramolecular method. The development of the collagen modification polyester, UMORFIL®T, can make a valuable contribution to the textile industry. After various experiments and characterizations, it was shown that the collagen modification polyester in this study contained a certain amount of collagen and retained the advantages of regular polyester. In addition, the characteristics of the original regular polyester were changed by the presence of collagen. The new properties included a natural champagne‑gold color, better hand-feel, and odor control, which make the material a premium and sustainable choice for functional textiles. This study creates a new direction for a circular economy product, collagen modification polyester, that can protect the ecology, reduce environmental pollution, protect planet soil, provide premium value for the textile industry, and raise the value of aquaculture.
Chapter
Byproducts from fish‐processing industries are rich sources of collagen, which could be extracted and further hydrolyzed by specific proteolytic enzymes to liberate biologically active peptides. The interest for the production of collagen‐derived biopeptides and their application in several fields, such as cosmetics, food additives, biomedical materials, and pharmaceuticals, has been increased during the past decade due to their wide range of biological activities, as well as their beneficial effects for humans. In fact, different activities have been attributed to collagen‐derived peptides including antioxidant, antihypertensive, antimicrobial, cholesterol‐lowering ability, hypoglycemic, anticancer, anti‐aging, antifreeze, etc. The present review focuses on the bioactive peptides derived from fish collagen byproducts. It describes the processes of collagen and gelatin extraction as well as collagen peptides (CPs) production. This review provides also an overview on the biological properties of collagen peptides as well as their potential applications in functional foods and pharmaceutical industries.
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Proteins from fish by-product sources are valuable source of bioactive peptides and show promise as functional foods ingredients. The objective of the present study was to isolate and characterize antibacterial peptides from protamex hydrolysates of Atlantic mackerel (Scomber scombrus) by-products. Four sequences SIFIQRFTT (P4), RKSGDPLGR (P8.1), AKPGDGAGSGPR (P8.2) and GLPGPLGPAGPK (P11) were identified in peptide fractions separated using RP-HPLC. At 200 μg mL(-1), while peptides P8.1, P8.2 and P11 exhibited partial inhibition, P4 totally inhibited tested Gram-positive (Listeria innocua) and Gram-negative (Escherichia coli) bacterial strains. These results suggest that the protein hydrolysate derived from mackerel by-products could be used as an antimicrobial ingredient in both functional food and nutraceutical applications. Copyright © 2015 Elsevier Inc. All rights reserved.
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A mixture of hexafluoroisopropanol–dichloromethane (1:4 v/v) acts as a fast, effective and convenient reagent for cleaving protected peptide fragments with a minimal amount of racemization from 2-chlorotrityl chloride resin.
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In molecular dynamics (MD) simulations the need often arises to maintain such parameters as temperature or pressure rather than energy and volume, or to impose gradients for studying transport properties in nonequilibrium MD. A method is described to realize coupling to an external bath with constant temperature or pressure with adjustable time constants for the coupling. The method is easily extendable to other variables and to gradients, and can be applied also to polyatomic molecules involving internal constraints. The influence of coupling time constants on dynamical variables is evaluated. A leap‐frog algorithm is presented for the general case involving constraints with coupling to both a constant temperature and a constant pressure bath.
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The PROCHECK suite of programs provides a detailed check on the stereochemistry of a protein structure. Its outputs comprise a number of plots in PostScript format and a comprehensive residue-by-residue listing. These give an assessment of the overall quality of the structure as compared with well refined structures of the same resolution and also highlight regions that may need further investigation. The PROCHECK programs are useful for assessing the quality not only of protein structures in the process of being solved but also of existing structures and of those being modelled on known structures.
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Phosphatidylethanolamine (PE) and phosphatidylglycerol (PG) are the main lipid components of the inner bacterial membrane. A computer model for such a membrane was built of palmitoyloleoyl PE (POPE) and palmitoyloleoyl PG (POPG) in the proportion 3:1, and sodium ions (Na+) to neutralize the net negative charge on each POPG (POPE-POPG bilayer). The bilayer was simulated for 25 ns. A final 10-ns trajectory fragment was used for analyses. In the bilayer interfacial region, POPEs and POPGs interact readily with one another via intermolecular hydrogen (H) bonds and water bridges. POPE is the main H-bond donor in either PEPE or PEPG H-bonds; PGPG H-bonds are rarely formed. Almost all POPEs are H-bonded and/or water bridged to either POPE or POPG but PE-PG links are favored. In effect, the atom packing in the near-the-interface regions of the bilayer core is tight. Na+ does not bind readily to lipids, and interlipid links via Na+ are not numerous. Although POPG and POPE comprise one bilayer, their bilayer properties differ. The average surface area per POPG is larger and the average vertical location of the POPG phosphate group is lower than those of POPE. Also, the alkyl chains of POPG are more ordered and less densely packed than the POPE chains. The main conclusion of this study is that in the PE-PG bilayer PE interacts more strongly with PG than with PE. This is a likely molecular-level event behind a regulating mechanism developed by the bacteria to control its membrane permeability and stability consisting in changes of the relative PG/PE concentration in the membrane.
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The fibrous collagens are ubiquitous in animals and form the structural basis of all mammalian connective tissues, including those of the heart, vasculature, skin, cornea, bones, and tendons. However, in comparison with what is known of their production, turnover and physiological structure, very little is understood regarding the three-dimensional arrangement of collagen molecules in naturally occurring fibrils. This knowledge may provide insight into key biological processes such as fibrillo-genesis and tissue remodeling and into diseases such as heart disease and cancer. Here we present a crystallographic determination of the collagen type I supermolecular structure, where the molecular conformation of each collagen segment found within the naturally occurring crystallographic unit cell has been defined (P1, a ≈ 40.0 Å, b ≈ 27.0 Å, c ≈ 678 Å, α ≈ 89.2°, β ≈ 94.6°, γ ≈ 105.6°; reflections: 414, overlapping, 232, and nonoverlapping, 182; resolution, 5.16 Å axial and 11.1 Å equatorial). This structure shows that the molecular packing topology of the collagen molecule is such that packing neighbors are arranged to form a supertwisted (discontinuous) right-handed microfibril that interdigitates with neighboring microfibrils. This interdigitation establishes the crystallographic superlattice, which is formed of quasihexagonally packed collagen molecules. In addition, the molecular packing structure of collagen shown here provides information concerning the potential modes of action of two prominent molecules involved in human health and disease: decorin and the Matrix Metallo-Proteinase (MMP) collagenase. • x-ray • fiber • crystallography • fibril • extracellular matrix
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Abstract Atlantic mackerel and its processing wastes were hydrolyzed separately using four commercial proteases, namely: Protamex, Neutrase, papain, and Flavourzyme. Mackerel and its by-products hydrolysates were rich in essential amino acids such as lysine, leucine and phenylalanine (3.44 − 7.81 mg g−1). Hydrolysis using protamex, neutrase or papain generated a high number of small peptides (300-700 Da). Both crude hydrolysates of mackerel and its by-products (1 mg mL−1) inhibited at least 40% of Listeria innocua HPB13 and Escherichia coli MC4100 growth, with crude by-products hydrolysate obtained by papain and neutrase being the most antilisterial with 69.7% and 65.2% of inhibition, respectively. Dose-dependent inhibition curves were obtained and complete inhibition of both L. innocua and E. coli was obtained after acetone fractionation of whole mackerel and its by-products hydrolyzed with protamex at 20 mg mL−1. Due to its high nutritional value, coupled with excellent functional properties mackerel by-products are of interest as functional foods.
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Horse purslane, a C4 species, is a branched, prostrate, and annual weed of upland field crops throughout the tropics. Experiments were conducted to determine the influence of various environmental factors on seed germination and seedling emergence of two populations of horse purslane. Seeds were collected from rice fields of the International Rice Research Institute (the IR population) and from sorghum fields of the University of the Philippines (the UP population); the two sites were 5 km apart in Los Baos, Philippines. Germination response of both populations was greater at 30/20 C and35/25 C day/night temperatures than they were at 25/15 C alternating day/night temperatures. Germination of both populations was greater in the light/dark regime than in darkness. In dark, depending on the temperature, seed germination of the UP population ranged from 37 to 62%, whereas seed germination of the IR population was <20%. Exposure to 5 min at 117 and 119 C for the IR and UP populations, respectively, reduced germination to 50% of maximum germination. Osmotic potential of-0.26 MPa inhibited germination to 50% of the maximum for the UP population, whereas the corresponding value for the IR population was-0.37 MPa. Seeds placed on or near the soil surface had maximum emergence, and emergence declined with increase in seed burial depth. Seedling emergence of the UP and IR populations was 74% and 13%, respectively, for seeds placed on the soil surface. For both populations, no seedlings emerged from a soil burial depth of 6 cm or more. Germination and emergence responses to light and seed burial depth differed between the two populations of horse purslane. Residues on the soil surface of up to 6 Mg ha -1 did not influence seedling emergence of either populations. Knowledge gained in this study could contribute to developing components of integrated weed management strategies for horse purslane.
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Our objective was to evaluate the angiotensin I converting enzyme (ACE) inhibitory activity of skate skin protein hydrolysates and its corresponding fractions. The skate skin hydrolysates were obtained by enzymatic hydrolysis using alcalase, α-chymotrypsin, neutrase, pepsin, papain, and trypsin. Amongst the six hydrolysates, the α-chymotrypsin hydrolysate had the highest ACE inhibitory activity compared to other hydrolysates. The amino acid sequences of the purified peptides were identified to be Pro–Gly–Pro–Leu–Gly–Leu–Thr–Gly–Pro (975.38 Da), and Gln–Leu–Gly–Phe–Leu–Gly–Pro–Arg (874.45 Da). The purified peptides from skate skin had an IC50 value of 95 μM and 148 μM, respectively, and the Lineweaver–Burk plots suggest that they act as a non-competitive inhibitor against ACE. Our study suggested that novel ACE inhibitory peptides derived from skate skin protein may be beneficial as anti-hypertension compounds in functional foods.
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Seed extracts of three plant species that grow wild in the arid regions of Tunisia, Juniperus phoenicea L. (Cupressaceae), Pistacia atlantica Desf. (Anacardiaceae), and Oudneya africana R. Br. (Brassicaceae), were examined for antimicrobial activity against bacterial food pathogens. Aqueous extracts were prepared and then precipitated with methanol or acetone. Extracted acetone fractions (pH 7.2) showed powerful antimicrobial activity, especially against Listeria monocytogenes, Listeria innocua, and Listeria ivanovii (Gram-positive) and were also active against Gram-negative strains Escherichia coli and Pseudomonas aeruginosa. Extracts selected for high antimicrobial activity were stable in the presence of organic solvents (chloroform, hexane, acetonitrile, methanol, and acetone), and withstand thermal treatments up to 100°C for 30 min. L. monocytogenes LSD530 and E. coli ATCC 25922 appeared to be inhibited by Juniperus and Pistacia extracts with a minimum concentration of 1.56 and 3. 12 mg/mL, respectively. This study established the potential of medicinal plants growing wild in arid regions of Tunisia as a source of antimicrobial agents.
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CHARMM27 is a widespread and popular force field for biomolecular simulation, and several recent algorithms such as implicit solvent models have been developed specifically for it. We have here implemented the CHARMM force field and all necessary extended functional forms in the GROMACS molecular simulation package, to make CHARMM-specific features available and to test them in combination with techniques for extended time steps, to make all major force fields available for comparison studies in GROMACS, and to test various solvent model optimizations, in particular the effect of Lennard-Jones interactions on hydrogens. The implementation has full support both for CHARMM-specific features such as multiple potentials over the same dihedral angle and the grid-based energy correction map on the , ψ protein backbone dihedrals, as well as all GROMACS features such as virtual hydrogen interaction sites that enable 5 fs time steps. The medium-to-long time effects of both the correction maps and virtual sites have been tested by performing a series of 100 ns simulations using different models for water representation, including comparisons between CHARMM and traditional TIP3P. Including the correction maps improves sampling of near native-state conformations in our systems, and to some extent it is even able to refine distorted protein conformations. Finally, we show that this accuracy is largely maintained with a new implicit solvent implementation that works with virtual interaction sites, which enables performance in excess of 250 ns/day for a 900-atom protein on a quad-core desktop computer.
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Research on fish composition has demonstrated that they constitute a promising source of molecules with health benefits. In the present study, Atlantic mackerel (Scomber scombrus) was processed using Protamex® and the distribution of nutrients in various fractions obtained following membrane filtration was analysed. The fish starting material was approximately 84% water and the dry matter contained 42.7% proteins, 45.2% lipids and 5.6% minerals. The recovery of fish dry matter in the liquid hydrolysate was 77.8%. Most of the fractions were protein-enriched and characterised by a well-balanced amino acid composition, notably in terms of the most essential amino acids and by molecules of relatively low molecular weight (≤42 kDa). Even before process optimising, the biochemical and nutritional analyses indicate that the Atlantic mackerel may provide high-value products for future applications in the health and food sectors.
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9-Fluorenylmethoxycarbonyl (Fmoc) amino acids were first used for solid phase peptide synthesis a little more than a decade ago. Since that time, Fmoc solid phase peptide synthesis methodology has been greatly enhanced by the introduction of a variety of solid supports, linkages, and side chain protecting groups, as well as by increased understanding of solvation conditions. These advances have led to many impressive syntheses, such as those of biologically active and isotopically labeled peptides and small proteins. The great variety of conditions under which Fmoc solid phase peptide synthesis may be carried out represents a truly "orthogonal" scheme, and thus offers many unique opportunities for bioorganic chemistry.
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Circular Dichroism (CD) relies on the differential absorption of left and right circularly polarised radiation by chromophores which either possess intrinsic chirality or are placed in chiral environments. Proteins possess a number of chromophores which can give rise to CD signals. In the far UV region (240-180 nm), which corresponds to peptide bond absorption, the CD spectrum can be analysed to give the content of regular secondary structural features such as alpha-helix and beta-sheet. The CD spectrum in the near UV region (320-260 nm) reflects the environments of the aromatic amino acid side chains and thus gives information about the tertiary structure of the protein. Other non-protein chromophores such as flavin and haem moieties can give rise to CD signals which depend on the precise environment of the chromophore concerned. Because of its relatively modest resource demands, CD has been used extensively to give useful information about protein structure, the extent and rate of structural changes and ligand binding. In the protein design field, CD is used to assess the structure and stability of the designed protein fragments. Studies of protein folding make extensive use of CD to examine the folding pathway; the technique has been especially important in characterising molten globule intermediates which may be involved in the folding process. CD is an extremely useful technique for assessing the structural integrity of membrane proteins during extraction and characterisation procedures. The interactions between chromophores can give rise to characteristic CD signals. This is well illustrated by the case of the light harvesting complex from photosynthetic bacteria, where the CD spectra can be analysed to indicate the extent of orbital overlap between the rings of bacteriochlorophyll molecules. It is therefore evident that CD is a versatile technique in structural biology, with an increasingly wide range of applications.
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A series of engineered linear analogues [coded as F6, W6, Y6, A6, S6 and C(Acm)6] were modeled, designed, synthesized and structurally characterized by mass spectra, circular dichroism, hydrophobicity analysis and molecular modeling. We have screened antimicrobial activity, hemolysis to rabbit erythrocytes, and cytotoxicity to human conjunctival epithelial cells. No significant hemolytic effect was observed for hBD3 or from five of the six analogues [F6, Y6, A6, S6 and C(Acm)6] over the range of 3-100 microg mL(-1). The six linear analogues have reduced cytotoxicity to human conjunctival epithelial cells over the range of 6-100 microg mL(-1) compared to hBD3. By tuning the overall hydrophobicity of linear hBD3 analogues, reduced cytotoxicity and hemolysis were obtained while preserving the antimicrobial properties. The decreased cytotoxicity of the linear analogues is suggested to be structurally related to the removal of disulfide bridges, and the flexible structure of the linear forms, which seem to be associated with loss of secondary structure. These results suggest a new approach for guiding the design of new linear analogues of defensin peptides with strong antibiotic properties and reduced cytotoxicity to mammalian cells.
Functional characterization of a protein sequence is a common goal in biology, and is usually facilitated by having an accurate three-dimensional (3-D) structure of the studied protein. In the absence of an experimentally determined structure, comparative or homology modeling can sometimes provide a useful 3-D model for a protein that is related to at least one known protein structure. Comparative modeling predicts the 3-D structure of a given protein sequence (target) based primarily on its alignment to one or more proteins of known structure (templates). The prediction process consists of fold assignment, target-template alignment, model building, and model evaluation. This unit describes how to calculate comparative models using the program MODELLER and discusses all four steps of comparative modeling, frequently observed errors, and some applications. Modeling lactate dehydrogenase from Trichomonas vaginalis (TvLDH) is described as an example. The download and installation of the MODELLER software is also described.
  • J Lee
  • X Cheng
  • J M Swails
  • M S Yeom
  • P K Eastman
  • J A Lemkul
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  • J C Jeong
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  • D A Case
  • C L Brooks
  • A D Mackerell
  • J B Klauda
  • W Im
  • Charmm-Gui Input Generator For
  • Namd Gromacs
  • Amber Openmm
J. Lee, X. Cheng, J.M. Swails, M.S. Yeom, P.K. Eastman, J.A. Lemkul, S. Wei, J. Buckner, J.C. Jeong, Y. Qi, S. Jo, V.S. Pande, D.A. Case, C.L. Brooks, A.D. MacKerell, J.B. Klauda, W. Im, CHARMM-GUI input generator for NAMD, GROMACS, AMBER, OpenMM, and CHARMM/OpenMM Simulations using the CHARMM36 additive force field, J. Chem. Theory Comput. 12 (1) (2016) 405e413.
The PyMOL Molecular Graphics System, Version 1.3r1
  • L L C Schrodinger
L.L.C. Schrodinger, The PyMOL Molecular Graphics System, Version 1.3r1, 2010.
  • M C Omez-Guill En
  • B Gim Enez
  • M E Caballero
  • M P Montero
M.C. G omez-Guill en, B. Gim enez, M.E. L opez-Caballero, M.P. Montero, Functional and bioactive properties of collagen and gelatin from alternative sources: a review, Food Hydrocoll. 25 (2011) 1813e1827.