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Characterization, amino acid composition and in vitro digestibility of hemp ( Cannabis sativa L.) proteins
Abstract
The protein constituents and thermal properties of hemp (Cannabis sativa L.) protein isolate (HPI) as well as 11S- and 7S-rich HPIs (HPI-11S and HPI-7S) were characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and different scanning calorimetry (DSC), and their amino acid composition and in vitro digestibility were also evaluated, as compared to soy protein isolate (SPI). SDS-PAGE analysis showed that the edestin (consisting of acidic and basic subunits, AS and BS) was the main protein component for HPI and HPI-11S, while HPI-7S was composed of the BS of edestin and a subunit of about 4.8kDa. DSC analysis characterized thermal transition of the edestin component and the possible present form of different subunits. Except lysine and sulfur-containing amino acids, the essential amino acids of various HPIs met the suggested requirements of FAO/WHO for 2–5year old infants. The proportion of essential amino acids to the total amino acids (E/T) for HPI (as well as HPI-11S) was significantly higher than that of SPI. In an in vitro digestion model, various protein constituents of various HPIs were much easily digested by pepsin plus trypsin, to release oligo-peptides with molecular weight less than 10.0kDa (under reduced condition). Only after pepsin digestion, in vitro digestibility of HPIs was comparable to that of SPI, however after pepsin plus trypsin digestion, the digestibility (88–91%) was significantly higher than that (71%) of SPI (P
... The results showed that HPA and HPAU mainly consisted of 15, 25, and 50 kDa protein constituents, whereas the HPS and HPSU were mainly consisted of 10-15, 20, and 35 kDa protein constituents, respectively (Fig. 1d), indicating that the protein extraction conditions affected the protein composition or quality. In particular, edestin is composed of six identical subunits, and each subunit consists of an acidic (AS) and a basic (BS) subunit linked together by a disulfide bond; AS is characterized by a molecular weight of 33-34 kDa, and the BS consists of two subunits of approximately 20 and 18 kDa, respectively [27,28]. In addition, the edestin fraction was identified as salt-soluble [27], indicating that the electrophoresis bands corresponding to molecular weights of approximately 20 and 33 kDa in HPS and HPSU were presumably the components of edestin protein fractions, one AS subunit, and two BS subunits, respectively, which is similar to previous findings. ...
... In particular, edestin is composed of six identical subunits, and each subunit consists of an acidic (AS) and a basic (BS) subunit linked together by a disulfide bond; AS is characterized by a molecular weight of 33-34 kDa, and the BS consists of two subunits of approximately 20 and 18 kDa, respectively [27,28]. In addition, the edestin fraction was identified as salt-soluble [27], indicating that the electrophoresis bands corresponding to molecular weights of approximately 20 and 33 kDa in HPS and HPSU were presumably the components of edestin protein fractions, one AS subunit, and two BS subunits, respectively, which is similar to previous findings. Both HPA and HPAU contained a protein constituent of approximately 45 kDa, corresponding to the subunit of soy β-conglycinin, which was in agreement with observation over hemp seed protein isolates extracted under alkaline conditions reported previously [28,29]. ...
... Protein molecules can undergo conformational alterations that may reveal previously enclosed amino acid side chains, allowing them to interact with water [40]. The balance between the water-attracting and waterrepelling properties of amino acids in proteins, along with factors like the pH and ionic strength of the environment, and the ability of polar amino groups in proteins to dissolve in water, which are crucial for protein-water interactions, are all vital [27]. The effects of the extraction methods on the WHC were evaluated in this study, and the results displayed in Fig. 3f show that the WHC of HPA and HPAU were markedly higher than those of HPS and HPSU. ...
... The hemp seed protein comprises of 75% (w/w) of salt-soluble edestin and 25% (w/w) of water-soluble albumin and rich in arginine contributing to their nutritional value [1] but deficient in the lysine and tryptophan [2]. The acidic subunit of hemp protein is quite homogeneous and has a molecular weight of around 34.0 kDa, whereas the basic subunit is largely composed of components with molecular weights of about 20.0 and 18.0 kDa [3], Most of 7S-rich components are essential subunits, which have a molecular weight of 4.8 kDa. The by-products of oil processing from hemp seeds is referred to as hemp seed meal (HSM) which contain protein content of 50-60% (w/w) [4]. ...
... It showed improved concentration of glutamic acid (18.3 mg/g), arginine (9.51 mg/g), aspartic acid (10.2 mg/g), methionine (1.63 mg/g), and lysine (5.9 mg/g). Similar amino acid profile was observed for hemp protein in a previous study [3]. ...
... The protein digestibility for HSM, HPH, and blend (1:1) were 68.2, 89.5, and 82.4% respectively. Wang et al. [3] also reported similar digestibility of 89-91% for trypsin and pepsin treated proteins. Moreover, blending of PPI and HPH increased the digestibility of PPI. ...
Plant-based proteins show poor functional properties and when a balanced source of amino acids is required, it cannot be provided only by a single source of plant-based protein. This study aims to enhance the functional and digestibility properties of hemp seed meal (HSM) protein by hydrolyzation (HPH) through ultrasonication and enzymatic treatment. The blending of HPH with pea protein isolate (PPI) was conducted to improve the amino acid profile. During ultrasonication, the protein content was significantly affected (p < 0.05) by ultrasonication time and solid-to-solvent ratio but not by the amplitude. Degree of hydrolysis was significantly (p < 0.05) affected by all three variables. Optimal hydrolysate was prepared by ultrasonication at 80% amplitude for 10 min and solid-to-solvent ratio of 1:30, with a protein content of 64.57% and a degree of hydrolysis (DH) of 6.25%. Under these conditions, papain treated HPH exhibited an increase in protein concentration (84.2%) and DH (17.3%). The blend of HPH and PPI in the ratio of 1:1 showed improved protein content (85.5%) with significant (p < 0.05) increase in solubility (72.8% at pH 7). This blend showed a good emulsifying activity index (33.4 m²/g) emulsifying stability (35.3 min), oil holding capacity (2 g/g), water holding capacity (1.8 g/g), foaming capacity (195.8%), and foaming stability (85.2%). This blend also displayed a balanced amino acid profile, with methionine and lysine contents of 1.63 mg/g and 5.88 mg/g, respectively. The blend of PPI:HPH (1:1) showed integration of HPH into the PPI matrix, due water bridging between protein particle as observed by scanning electron microscopy (SEM). The amide I at 1627.6 cm⁻¹ in HSM shifted to 1633.7 cm⁻¹ in HPH and amid II at 1518.7 cm⁻¹ in HSM shifted to 1535.3 cm⁻¹ in HPH indicating a change in secondary structure of HSM due to ultrasonication and enzymatic hydrolysis. In Vitro protein digestibility of 1:1 blend was also higher compared to other samples. The blend of HPH and PPI blend have good potential for creating nutritionally enhanced and functionally superior plant-based food products.
... SDS-PAGE electropherograms were generated using a Gel Doc™ EZ Imager (Bio-Rad, Mississauga, ON, Canada) and analysed with Image Lab (version 4.1, Bio-Rad). The relative protein content for the major bands (subunit or protein band) was semi-quantified following the Foods 2024, 13, 531 7 of 20 procedure described by Wang et al. (2008) [36] based on the relative area of each band in relation to the total area. Samples were measured in duplicate. ...
... SDS-PAGE electropherograms were generated using a Gel Doc™ EZ Imager (Bio-Rad, Mississauga, ON, Canada) and analysed with Image Lab (version 4.1, Bio-Rad). The relative protein content for the major bands (subunit or protein band) was semi-quantified following the Foods 2024, 13, 531 7 of 20 procedure described by Wang et al. (2008) [36] based on the relative area of each band in relation to the total area. Samples were measured in duplicate. ...
... As shown in Figure 5A, all varieties exhibited three major bands of 15 to 35 kDa (marked as the letters b, c and d) and one minor band of about 45kDa (marked as a). Previous studies have also shown the presence of three major bands corresponding to acidic and basic subunits (AS and BS) of edestin [13,36,63]. These authors reported that the AS is about 30-35 kDa, and the BS consists of two subunits of about 15-20 kDa, respectively [36,64]. ...
The increasing use of hempseed in food products highlights the need for a comprehensive database for scientific research and industrial applications. In food development, information about the techno-functional properties of raw materials plays a crucial role in determining the suitability of each product for specific applications. Thus, this study aims to characterise three hempseed varieties (Ferimon, Henola and Uso-31), comparing their physicochemical and nutritional compositions. Moreover, the study investigates the impact of hempseed varieties on the techno-functional, physical and thermal properties of the partially defatted hempseed flours (PDHFs) obtained from single screw pressing (SSP) oil extraction. The fatty acid and tocopherol profiles of the dehulled seeds and oil were also analysed. Significant variations in yield and physical properties were observed among hempseed varieties, influenced by genetics, adaptation to agro-climatic conditions and cultivation systems. Despite its lower yield (kg/ha), Uso-31 exhibited superior 1000-seed weight, dehulling yield and larger mean seed size (1.79 ± 0.02 mm). Hempseed oil was rich in unsaturated fatty acids, particularly linoleic (51.2–53.4 g/100 g oil) and α-linolenic (14.88–18.97 g/100 oil) acids, showing variations in γ- and α-tocopherols depending on the variety. The variety also influenced the least gelation concentration (LGC) and techno-functional properties such as water absorption capacity (WAC), emulsifying activity (EA) and emulsion stability (ES). SDS-PAGE and DSC measurements indicated the presence of 11S and 7S globulin proteins with denaturation temperatures above 87.8 °C. These findings confirm that the studied hempseed flours are valuable techno-functional and nutritional ingredients suitable for sustainable food formulations.
... In fact, soybeans are considered the main source of vegetable proteins, with a composition very close to that of animal-origin food. It should be noted that while soybeans contain anti-nutritional factors, such as trypsin inhibitors requiring thermal treatment for elimination, hemp contains a smaller amount, making its proteins more digestible [20]. The significant differences shown in Table 1 are related to the different methods used for determination. ...
... The apparent digestibility of GE values obtained appears high. [20] and [21] report that hemp flour has high digestibility related to a high degree of digestibility of proteins. [20] reports that this good digestibility can be linked to an immediate release of bio accessible amino acids. ...
... [20] and [21] report that hemp flour has high digestibility related to a high degree of digestibility of proteins. [20] reports that this good digestibility can be linked to an immediate release of bio accessible amino acids. ...
Aims: The aim of this study was to evaluate the utility of NIRS for predicting the energy content through the chemical characterization of the flour obtained after the cold pressing of Cannabis sativa L. seeds, as well as the possibility of predicting their energy content starting from the data obtained through the NIRs technique. Study Design: The chemical composition of 56 hemp flour samples was determined following the official protocols of the Association of Analytical Chemists and chemometric readings were conducted. GE, gross energy digestibility (GEd) and digestible energy (DE) were estimated using the equations proposed by INRA. A statistical analysis was performed to evaluate the potential use of NIR data to predict the energy content of hemp flour. Results: Data from laboratory and NIR assessments were 22.54 versus 20.44 for GE (MJ/kg DM), 90.72 versus 90.21 for GEd (MJ/kg DM), and 19.73 versus 20.13, respectively for the loss (%). The results indicated the feasibility of energy value prediction, although further studies are needed to refine the technique. NIR expands the calibration set, allowing increasingly accurate determinations, in the study of the chemical-nutritional characteristics of hemp sativa, even if further investigations are necessary. Conclusion: The study provides comprehensive insights into the chemical composition of hemp flour, explores its comparison with other seeds, evaluates different analysis methods, and establishes reliable prediction models for energy content.
... In comparison to soybean, all EAAs were higher (Figure 7). In fact, lysine would be the first limiting amino acid in hempseeds (2.13 vs. 0.85 g/100 g seed) that has been already stated in the literature [20,[42][43][44]. Both hempseed and soybean are similarly low in sulfurcontaining amino acids. ...
... In comparison to soybean, all EAAs were higher (Figure 7). In fact, lysine would be the first limiting amino acid in hempseeds (2.13 vs. 0.85 g/100 g seed) that has been already stated in the literature [20,[42][43][44]. Both hempseed and soybean are similarly low in sulfur-containing amino acids. ...
Hemp is considered one of the potential novel crops for human and animal nutrition. This study aimed to determine the complete chemical composition of 29 different varieties of whole hempseeds. Fatty acid composition, amino acid profile, mineral composition, and cannabinoids content were also evaluated. All hempseed varieties were milled to obtain whole hempseed flour. Differences between hempseed varieties were significant (p < 0.05) for all measured parameters. Proximate composition results showed that crude protein and fat contents varied from 21.6–28.9% and 21.1–35.7%, respectively. Fatty acid profiles revealed that the three major fatty acids were linoleic acid (52.79–57.13%) followed by α-linolenic acid (12.62–20.24%), and oleic acid (11.08–17.81%). All essential amino acids were detected in all varieties, with arginine (12.66–17.56 mg/100 g protein) present in abundance, whereas lysine was limiting. Substantial differences were found in the mineral content, and potassium (509.96–1182.65 mg/100 g) and iron (5.06–32.37 mg/100 mg) were the main macro- and microminerals found. All cannabinoids were found in small traces and tetrahydrocannabidiol (THC) was only detected in five varieties. To conclude, the nutritional composition of hempseeds with hull makes them suitable to be added into the diets of humans or animals as a highly beneficial novel ingredient.
... Hemp protein contains all nine essential amino acids, generally with high levels of glutamine and arginine (Leonard et al., 2020). The amino acid profile is comparable to that of soy, but with better enzyme digestibility (Wang et al., 2008). Given its rich amino acid profile, hemp seed is proposed as a good matrix for the development of functional food products with the nutritional claims "source of/high in" protein. ...
... The amino acid profile of Futura 75 (Table 3) hemp seed consistent with previous research on different varieties (Callaway, 2004;Wang et al., 2008). ...
Current trends in the snack bar market emphasize the incorporation of alternative protein sources while simultaneously addressing the criteria of being both “high in fiber” and “high in protein”. Hemp seed, the seed of non-psychoactive Cannabis Sativa L, stands out as a significant source of protein, dietary fiber, minerals, and unsaturated fats. This study aims to explore the nutritional potential of hemp seed to develop a functional food that responds to the needs of such a market highly demanding of plant-based alternatives. Along with seeds protein nutritional quality analysis and lipid profile characterization, three snack bar samples containing hemp seed as a functional ingredient were formulated, according to nutritional claims regulations. Two products with 20% hemp met sensory acceptability criteria. This study suggests hemp seed as a potential functional food ingredient to meet the demand for plant-based alternatives, offering quality protein, digestive benefits due to its high fiber content and an optimal omega 6 to omega 3 ratio. However, there appears to be an upper limit for hemp seed in product formulations due to sensory issues. This study showed that adding more than 40% to a snack bar significantly reduced consumer acceptability and purchase intent.
... Hemp seeds, derived from the mature seeds of Cannabis sativa L., a member of the Cannabaceae family [1], originated from China and were later cultivated in many regions across Europe, South America, Africa, and other countries [2]. Hemp seeds, which are recognized as one of the first groups of "medicinal and edible" traditional Chinese herbs, are abundant in a variety of nutrients and exhibit numerous physiological activities [3]. ...
Hemp seeds are a rich source of protein with potential health benefits, and the extraction and purification of bioactive peptides from these can make the development of novel functional food ingredients and therapeutic agents. Single-factor and response surface experiment were carried out with the yield of peptides as the main determination index to identify the optimal process conditions. The highest peptides yield, which was 66%, was achieved with the 2% enzyme addition, the temperature of 60 °C, and the duration of 4 h. Next, a component with enhanced activities, named HSP4-1, was prepared from the hydrolysate after ultrafiltration and Sephadex G-15 chromatography. HSP4-1 exhibited significant antioxidant activity, as evidenced by its scavenging effects on DPPH, hydroxyl radicals, and ABTS, with IC50 values of 0.19, 0.14, and 0.52 mg·mL⁻¹, respectively. Additionally, it demonstrated α-glucosidase inhibitory activity, with an IC50 of 0.17 mg·mL⁻¹. The sequences of the peptides were subsequently determined by high-performance liquid chromatography-mass spectrometry (HPLC–MS), as AGFLGVDEFR, AGLFNSR, and LAFDR, which are hypothesized to contribute to the bioactivities. The identified peptides have the potential to serve as raw materials for natural antioxidants and hypoglycemic drugs. This not only facilitates their application in the development of functional foods and pharmaceuticals but also offers valuable process parameters and theoretical support for the deep processing and high-value utilization of hemp seeds.
... Additionally, given its substantial dietary fiber content (18.47%), the obtained HPC can further enrich the nutritional profile of choux pastry. According to the literature sources, hemp protein demonstrates superior bioavailability, with digestibility rates between 88% and 91% [52], significantly outpacing that of traditional sources like soybean, which shows digestibility rates ranging from 20.58% to 50.21% depending on the processing method [53]. Additionally, Liener (1994) notes that while soybeans possess high nutritional value, they also contain anti-nutritional factors such as trypsin and chymotrypsin inhibitors, which impede protein digestion and overall nutrient absorption in the intestine [54]. ...
This study explores the production and valorization of hemp seed cake protein concentrate (HPC) as a functional ingredient to enhance the nutritional quality and sensory attributes of choux pastry products, specifically éclairs. By integrating varied concentrations of HPC (0%, 1%, 5%, 10%, 15%, and 20%) into traditional formulations, the physicochemical properties, proximate composition, amino acid profile, and sensory characteristics of the resulting pastries were assessed. Sensory attributes were assessed using the check-all-that-apply (CATA) method, where a trained panel selected applicable descriptors from a predefined list. Results indicated that the incorporation of HPC significantly increased protein content from 8.23% in the control sample (HPC0%) to 11.32% in the HPC20% formulation and improved moisture retention, leading to greater exterior and interior éclairs volume, increasing from 42.15 cm³ to 51.5 cm³ and from 18.34 cm³ to 38.47 cm³, respectively. Furthermore, sensory evaluation revealed pronounced differences in attributes such as flavor, appearance, and mouthfeel, with optimal sensory profiles noted at 10% HPC inclusion. The amino acid analysis demonstrated a balanced composition, particularly of essential amino acids, emphasizing HPC’s potential as a valuable protein source, with significant contributions from leucine (8.17 g/100 g protein), isoleucine (5.56 g/100 g protein), and phenylalanine (6.31 g/100 g protein), as well as notable levels of immunoactive amino acids such as arginine (10.92 g/100 g protein) and glutamic acid (20.16 g/100 g protein). These findings highlight the significant nutritional benefits of HPC enrichment, supporting the development of healthier bakery products and contributing to sustainable food practices within the industry.
... What distinguishes this food source from other plant-based foods is the presence of all the essential amino acids required by humans and high levels of arginine, which plays a dual role in immune regulation by modulating the inflammatory response [8], and glutamine, which supports intestinal epithelium integrity and immune cell function [9][10][11]. In addition, hemp seed proteins are less allergenic, making it a safer option for people sensitive or prone to food-related immune reactions [12] and easier to digest than soy protein [13]. The hydrolysis of hempseed proteins results in hydrolysates that exert different biological activities, such as antioxidant [14,15], antihypertensive [16,17], and anti-inflammatory properties [17,18]. ...
Hempseeds, from the Cannabis sativa plant, and its derivates are a versatile food option for various dietary preferences. Due to their aminoacidic profile, researchers have studied the presence of bioactive peptides in hempseed proteins. In this study, the water-soluble fraction of hempseed protein was extracted, and the derived peptides were analyzed. The investigation focused on their biological function, particularly their antioxidant activity. Several biological functions have arisen, such as angiotensin-converting enzyme inhibition activity, dipeptidyl-peptidase IV, dipeptidyl-peptidase III inhibition, and ubiquitin-mediated proteolysis activation. The hydrolysates show greater 2,2-azinobis-[3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical scavenging activity compared to the proteins (97.95 ± 4.48 versus 81.04 ± 10.63). Furthermore, the impact of these proteins and peptides on the U937 cell line was evaluated to assess cell viability and their potential role in modulating inflammation associated with gastrointestinal autoimmune diseases. Protein treatment resulted in a significant reduction in cell viability, as opposed to hydrolysates, which did not affect it.
... Globulins are generally more susceptible to hydrolysis by digestive enzymes [40]. Furthermore, it has been reported that the 7S and 11S fractions of globulins exhibit different in vitro digestibility profiles [41]. ...
This review aims to understand the techno-functional and structural properties of bean proteins, highlighting their strengths and weaknesses while presenting them as a robust alternative protein source with high potential to become a competitive ingredient in the protein market. For this purpose, ScienceDirect and Scopus were used as databases with the keywords “bean proteins”, “protein modifications + beans”, and “techno-functional properties + beans” to consult the relevant literature. This could reduce global dependence on soy and pea proteins. The study compiles various current articles that address desirable techno-functional properties and potential modifications for a wide range of food industry applications. Based on the gathered findings, bean-derived proteins exhibit a more hydrophobic nature and a more compact structure compared to soy and pea proteins. Consequently, they demonstrate superior emulsifying properties and an excellent oil absorption capacity, making them promising ingredients for emulsified products and baked goods. On the other hand, soy and pea proteins perform better in meat-based products and confectionery due to their higher water absorption capacity and good stability.
... Hemp seed, in particular, has gained attention as a "superfood" due to its high protein content, which accounts for 20-26% of unhulled seeds (Callaway 2004;Potin and Saurel 2020). Hemp proteins contain all nine essential amino acids and exhibit excellent digestibility (Wang et al. 2008). Notably, hemp seeds are rich in arginine, which makes up about 12% of their amino acid profile, compared to less than 7% in most food-based proteins (Callaway 2004). ...
In Europe, hemp fiber has historically been utilized in textile, paper, and construction industries prior to the emergence of synthetic fibers. The demand for hemp fibers in the European Union (EU) has led to a significant increase in cultivation area, rising by 46.5% from 22,010 hectares (ha) in 2016 to 32,250 ha in 2022. Recently, the European Parliament relaxed hemp regulations by raising the allowable THC level from 0.2% to 0.3%. France stands as the largest hemp fiber producer, contributing 78% of EU production, equivalent to 121,720 tonnes in 2022. This increase in production is driven by the fibers’ use in textiles, clothing, paper, and hemp seed in food products. Notably, the building industry has seen a rise in the use of hemp, particularly in materials such as hemp insulation and hemp concrete. Furthermore, innovations in hemp include the development of bioplastics and the replacement of glass fibers with hemp fibers in the automotive sector. This review explores the regulatory landscape, industrial applications of hemp fiber, and the future potential of hemp by-products as alternative agricultural commodities in EU countries.
... The in vitro digestion process and protein digestibility determination were performed according to the method described by Wang et al. [37]. Protein digestibility was calculated based on the difference between the introduced protein and the protein determined in the sample after the in vitro digestion process. ...
The aim of this research was to develop a technology for the production of plant-based burgers (PBBs) based on potato protein, also containing high content of iron and appropriately selected fats. The produced PBBs were characterized in terms of their nutritional and bioactive properties both before and after the in vitro digestion process. It was found that the produced burger was characterized by high protein content, ranging from 20.80 to 22.16 g/100 g. It was also shown to have a high dietary fiber content, ranging from 8.35 to 9.20 g/100 g. The main fraction of dietary fiber in the tested samples was insoluble fiber, which accounted for approximately 89% of the total fiber content. In addition, noteworthy is the high digestibility of the protein, reaching approximately 95% for the potato fiber used in the formulation, and about 85% for the oat fiber. Produced PBBs also provide significant amounts of iron, with the use of an organic iron source greatly increasing its quantity in the final product. The analyzed antioxidant properties before and after the digestion process showed a tenfold increase in biological activity after digestion, indicating that the examined PBBs may counteract oxidative stress. Analyzing the chemical and biological properties, it is impossible not to assess consumer attractiveness. It has been shown that PBB1, which contains potato fiber and powdered sprouts enriched with ferritin, received the highest attractiveness ratings among respondents.
... One of the advantages of hemp protein compared to other vegetable proteins is its easy digestibility, which is about 88-91 % [31], therefore, studies into the effectiveness of its use in meat products technologies are needed. ...
The object of this study was the technology of cooked sausages with hemp seed protein. The research considers determining the effectiveness of using protein from hemp seeds in the technology of cooked sausages to increase the nutritional value and improve the functional and technological indicators when replacing part of the meat raw material. The subject of the research was meat model systems, cooked sausage with hemp seed protein. Three prototypes of cooked sausages based on the analog were developed. In experimental samples of cooked sausages, second-grade beef was replaced with protein from hemp seeds (Cannabis Sativa L.) (ToV "Desnaland", Ukraine) in amounts of 12, 14, and 16 %. The functional and technological indicators of hemp seed protein were previously investigated at hydromodules of 1:1, 1:2, and 1:3. It was found that hydrated hemp seed protein in a ratio of 1:1 had the best parameters for introduction into minced meat systems. It was determined that the introduction of proteins from hemp seeds (hydromodule 1:1) in the amount of 12–16 % increases the water-binding capacity of minced meat to 97.8 %, plasticity by 16.19–23.85 %, and the content of total moisture in samples of cooked sausages after heat treatment by 5.08–7.08 %, yield of the finished product up to 129.83 %. Replacing second-grade beef in cooked sausage with protein from hemp seeds causes an increase in the mass fraction of protein in finished products by 14.99–19.98 %, and mineral substances by 68.52–97.22 %. The organoleptic indicators of the experimental cooked sausages met the regulatory requirements. The safety of the developed products was established according to microbiological indicators. It has been proven that the use of hydrated protein from hemp seeds in the technology of cooked sausages in the amount of 12–14 % makes it possible to obtain a meat product with a high level of functional and technological indicators of model meat systems and organoleptic indicators of finished products. The use of protein from hemp seeds in the production of cooked sausages makes it possible to expand the range of products popular among the population
... In modern times, the sativa subspecies is legally cultivated in many countries, not only for its seeds, an excellent source of lipids, proteins, carbohydrates, minerals, and vitamins, but also for the production of "Cannabidiol = CBD" (non-psychoactive cannabinoid), which is of significant importance (Ferrini et al. 2021). The existing literature primarily focuses on CBD, overlooking the biologically valuable molecules present in both the seeds and leaves of Cannabis sativa, which hold high nutritional value (Callaway 2004;Wang et al. 2008;Da Porto et al. 2012;Teh and Birch 2013;Pojic et al. 2014;Liang et al. 2015;Schluttenhofer and Yuan 2017;Mikulec et al. 2019;Rapa et al. 2019). ...
This study was conducted to evaluate the wound-healing activities of a Cannabis sativa L. plant extract and cannabidiol on incision wounds. An incision was created and sutured in rats under anaesthesia. Routine wound care procedures were applied for 10 days, followed by histological wound examinations. The cellular bioactivities of the hemp extract and CBD were assessed for MCP-1, EGF, BFGF, IL-8, and COL-1 using ELISA on the rat skin wound healing activity. A one-way ANOVA was used for the data analysis. The EGF values in the plasma were similar in the povidone-iodine, hemp seed oil, and hemp essential oil groups (P > 0.05). However, the EGF levels were lower in the CBD group compared to the other groups (P < 0.001, P < 0.005). The MCP-1 values in the hemp seed oil, hemp essential oil, and CBD were similar (P > 0.05), whereas povidone iodine exhibited lower MCP-1 levels compared to the other groups (P < 0.001, P < 0.005). It was determined that the plasma BFGF, IL-8, and COL 1 values of the groups were similar (P > 0.05). To our knowledge, this study is the first to evaluate the effects of CBD, seed oil, and hemp leaf extract on incision wound healing. It demonstrates that hemp extract holds greater potential benefits for wound healing compared to CBD.
... La fertilización es un factor agronómico importante en el cultivo de cáñamo (Amaducci et al., 2015), ya que posee gran impacto en el rendimiento y calidad de biomasa (Van Der Werf, 2002;Schumacher et al., 2020). Para el tipo de cannabis de fibra, la tasa de fertilización que se sugiere es de alrededor de 50 a 200 kg N/ha Ehrensing, 1998); sin embargo, es difícil estimar la necesidad en el caso del cannabis que tiene un uso medicinal o recreativo, debido a la diferencia de especies, las condiciones en los cultivos (Wright y Niemiera, 1987) y la demanda de nutrientes que se presentan en las etapas de crecimiento (Wang, 2008). En las figuras 19 a 21, se describe las deficiencias y excesos de los principales elementos NPK. ...
El cáñamo ha sido de gran importancia económica como proveedora de fibras,
alimento y 29 medicinas durante más de seis mil años, se puede señalar también
que se cultivaba en casi todos los países europeos y asiáticos (Ramos, 2019). El
cáñamo ha sido modificado desde años atrás, por esta razón es muy difícil saber a
ciencia cierta el origen geográfico, se discute su procedencia, pero algunos autores
coinciden en que es originaria de Asia Central, donde luego se expresando por todo
el mundo (Alonso et al., 2021). Los primeros usos que se le dieron a la planta de
cannabis están documentados en la India, el Medio Oriente, Grecia, Roma y en la
gran mayoría del continente asiático (Franco, 2019).
... The amino acid profile of hemp protein is comparable to high-quality protein sources such as egg white and soy proteins [81]. While the literature on the bioavailability of hemp protein is still scarce, the in vitro digestion of isolated hemp proteins showed greater digestibility than isolated soy proteins [82,83]. ...
This review discusses different protein sources and their role in human nutrition, focusing on their structure, digestibility, and bioavailability. Plant-based proteins, such as those found in legumes, nuts, and seeds, may contain anti-nutritional factors that impact their bioavailability apart from structural and compositional differences from animal proteins. Animal proteins are generally highly digestible and nutritionally superior to plant proteins, with higher amino acid bioavailability. Alternative protein sources are also processed in different ways, which can alter their structure and nutritional value, which is also discussed.
... Hemp seed protein has a nutritionally relevant amino acid content. Wang, Tang, Yang, and Gao (2008) reported that the essential amino acid content available in hemp seed is highly digestible and meets the recommended FAO/WHO (Food and Agriculture Organization/World Health Organization) requirements recommended for infants. The weight of total phenolic substances in hemp flour was 980.71 ± 21.93 mg GAE*kg-1, which is around 2.25 times more than wheat flour, while the antioxidant content weighed 1584.11 ...
In the present research, the use of hemp flour in gluten-free cake production for celiac patients was investigated. Samples were produced by adding hemp flour at varying ratios (25%, 50%, 75%, 100%) to the gluten-free flour mixture, and physical, chemical, and sensorial analyses were performed. The addition of hemp flour to the formulation resulted in a variation in ash content ranging from 3.50% to 7.08%, while the fat content varied between 22.64% and 37.04%. The dietary fiber content increased by 5.00% to 9.55%, and the protein content increased by 9.96% to 17.89%. Furthermore, the total phenolic content increased to 188.30 mg GAE/g, and the antioxidant content showed an increase ranging from 49.92% to 82.16%. Whereas the nutritional value of gluten-free cake samples increased with the use of hemp flour, adding over 50% to the formulation resulted in adverse effects on physical and sensorial properties. As a result of the research, it was concluded that hemp flour is usable in gluten-free cake samples in terms of increasing the nutritional value and obtaining a new product, but it should not be used at a rate of over 50%.
... The amino acids profile hasn't been described extensively yet. The main protein found in hemp genus plants is edestine, which is rich in valuable amino acids like glutamic acid and aspartic acid (Wang et al. 2008). Hemp seed protein is rich in sulphurcontaining amino acids, methionine and cystine (Odani et al. 1998) and has higher levels of arginine and glutamic acid than soybean meal (Callaway, 2004). ...
Initially, the nutritional value of the three main by-products of Cannabis sativa (hempseed cake, second-rate quality hemp seed and hemp hay), cultivated under Greek conditions and collected from four plantations in the Macedonian region was estimated. A second part of the study was the investigation of the effects of diet inclusion with hempseed cake (HSC) on the performance of Holstein dairy cows. The HSC used for this experiment was the one analyzed in the first part of the study. A total number of 20 lactating cows were allocated into two equal groups in a randomized block design. Cows of both groups were offered a total mixed ration (TMR) on the same feed allowance. The diet of the experimental group was formulated with 3.5 % hemp cake, at a quantity of 1kg of hemp cake per cow per day. Milk yield was recorded individually, and feed refusals were recorded on a pen basis daily, during the first 40 days of lactation. Individual milk samples were analyzed for their chemical composition, lipid oxidative stability and fatty acid composition and tetrahydrocannabinol (THC) and cannabidiol (CBD) content, as well as feed samples of each group. Individual blood samples were received for biochemical indices assessment. All data were subjected ANOVA statistical analysis. The results of hemp by-products chemical profile showed similar nutritional values among the different cultivation regions. The experimental results indicated that the inclusion of HSC did not affect milk production and composition (P > 0.05). However, diet inclusion with HSC favorably influenced milk fatty acid profile. Finally, serum NEFA concentration was lower for the HSC group while serum urea levels were higher (P < 0.05).
... The seeds contain valuable proteins, which exhibit an antioxidant effect due to their amino acid composition [Girgih, Udenigwe and Aluko 2013]. The protein composition of these seeds make them an excellent natural source of amino acids [Wang et al. 2008]. ...
The purpose of the study was to evaluate consumer awareness of the medicinal properties of hemp and its applications in various industries. Before commencing the survey, the following research hypothesis was made: the survey respondents would lack basic knowledge about hemp. The study group of survey respondents consisted of 50 randomly selected people from the Pomeranian Voivodeship, Poland. The research tool was an original survey questionnaire containing 11 questions and a section for sociodemographic data. The survey study was conducted from October to December 2018. As a result of the study, the research hypothesis was positively verified. It was found that most members of the study group were unfamiliar with the term hemp, but still associated it positively. This was related to the erroneous attribution of its properties to Cannabis indica. More than half of the study group were unable to state whether the plant in question was used in medicine. The level of knowledge among the study group concerning the applications of hemp in various industries was low.
... From the metabolomic point of view, our study confirmed hemp as an attractive non-food crop, that produces a wide variety of resources [56]. Building on this perspective, the advantages of incorporating hemp into industrial products are evident. ...
Agricultural diversification and high-quality products deriving from sustainable crops such as hemp can represent a solution to revitalize marginal areas and reverse land abandonment. This study aimed at comparing four different hemp cultivars (Carmagnola Selezionata, “CS”; Futura 75, “FUT”; Felina 32, “FEL”; Secuieni Jubileu, “JUB”) to provide information to select the best suited cultivar for cultivation in mountain marginal areas and for specific end-use applications. Hemp cultivars were cultivated in a single experimental field to compare their ecological and agronomic behavior (duration of life cycle phases, plant size and biomass allocation, and plant resource-use strategies). Through metabolomic analysis of both vegetative and reproductive parts of the plants we tested the presence of substances of nutraceutical interest and traced seed nutritional profile. The four cultivars had different ecological and agronomic behavior, and nutritional profile. We found several compounds with potential pharmaceutical and nutraceutical values in all parts of the plant (leaves, inflorescences, and stems). JUB resulted the most suitable for seed production while CS showed the highest content of bioactive compounds in flowers and leaves. FUT, showed the best suitability for multi-purpose cultivation, while FEL seemed to be not appropriate for the cultivation in mountain area. The multi-disciplinary approach we adopted was effective in distinguish across hemp cultivars and provided information to farmers for the selection of the best hemp cultivar to select. Hemp had a high potential for cultivation in marginal lands, demonstrating to be an economic resource due to its multi-purpose use and to the possibility to generate high-added values products. Our results could serve as a stimulus for the reintroduction of this culture in the study area and in other similar environments.
... Therefore an additional basic redispersing and centrifugation step of oleosomes was applied to remove these proteins. The SDS-page of the washed i.e., purified, oleosomes are shown in Fig. 1a, lane 2. Clearly, after the washing only the proteins at 15 KDa remained, which mostly like are the proteins that are absorbed on the oleosome membrane and cannot be removed with the washing step (Aluko, 2017;Garcia et al., 2021;Wang, 2008). ...
The interest in Cannabis sativa L. has been on the rise recently, driven by its potential applications in various sectors, including the food industry, the medical sector, and other key areas. This crop possesses a diverse profile of essential fatty acids and a range of bioactive compounds, which exhibit properties that are highly significant for functional food ingredients and nutraceutical purposes. The objective of this study was to investigate the characteristic lipid and bioactive profiles of different plant parts (e.g., inflorescences and leaves) to ascertain their possible uses in nutritional and therapeutic fields. The fat content of the plant material was determined by the Soxhlet method, and gas chromatography was employed for the assessment of the fatty acids and selected bioactive compounds profile. In addition, some lipid quality indices were calculated with the purpose of providing a more in-depth discussion of these aspects beyond the traditional n-6/n-3 ratio. A distinct lipid composition was evident among the various plant parts. Compared to inflorescence samples, leaves typically contain higher proportions of SFAs, MUFAs, PUFAs, and n-3 fatty acids, along with a more favorable n-6/n-3 ratio, which may significantly impact nutritional value. Phytol-rich leaves can suggest its potential application as a functional feed or even a nutraceutical. Furthermore, the occurrence of hexacosane and related antimicrobial and antifungal compounds serves to enhance the practical utility of the leaves. Notably, hemp leaves are not merely a by-product, but rather offer significant practical applications.
Hemp seed protein isolates (HPI) were used to produce a gel through probiotic fermentation. This study assessed how fermentation time (ranging from 0 to 16 h) affected the physicochemical properties of the HPI gel. The results indicated that gel formation began after 8 h of fermentation, as demonstrated by a pH decrease, an increase in particle size, and the development of aggregation observed through fluorescence and scanning electron microscopy. The gel produced after 16 h of fermentation showed the highest viscosity, storage modulus, and gel strength, attributed to stronger molecular interactions, including non-covalent and covalent crosslinking. However, the gel produced after 12 h of fermentation showed the highest water-holding capacity, and extending the fermentation beyond 12 h caused a decrease in water-holding capacity. Additionally, the subunits tended to form polymers after fermentation, suggesting that gel formation was influenced by both acidification and specific covalent crosslinking. These findings propose that HPI could serve as a viable alternative for developing plant-based gel products.
Industrial hemp ( Cannabis sativa L.) is a multifunctional raw material with a wide spectrum of applications. A huge interest in hemp has arisen in the food industry comparatively recently. Seeds and cake of hemp have the high protein content (20.00-38.70% and 27.90-40.70%, respectively), which makes them a promising raw material for production of concentrates, isolates, and hydrolysates for the food industry. Hemp cake is distinguished by the high content of fiber (17.41-60.38%) and can be used as a prebiotic component of food. In terms of the amino acid content, the amino acid composition of hemp exceeds the corresponding indicators of the reference protein recommended by WHO. The content of lysine is the only indicator, in which hemp protein is inferior to the “ideal” protein. Digestibility of protein of hulled hemp seeds is in a range from 90.8% to 97.5%, which is comparable to digestibility of casein. Digestibility of hemp isolate is 88-91%, which is 21.9% higher than that of soy protein isolate. Peptides and amino acids contained in hydrolysates of hemp protein can show the high biological activity. Hydrolysates attract interest among researchers not only because of their bioactivity but also because of their high digestibility and nutritional value. Targeted proteolysis is a tool that facilitates an improvement in the functional-technological properties of protein. Seeds and cake are a promising raw material for using in food technologies to produce vegetable oil, dietary fiber, protein preparations and functional products. The aim of this paper is to analyze the main methods for industrial hemp processing and promising directions of using protein products from waste of hemp oil production in the food industry.
Hempseed meal, a co-product of hemp oil extraction, contains about 40 % proteins, which has recently become abundant because of the large demands of hemp oil related products. It has been an acute issue for hemp industries to utilize the hempseed meal for value-added products. On the other hand, wood industries are constantly seeking plant protein alternatives not only to secure plant protein sources but also to improve wet adhesion properties. In this study, native hempseed protein was identified to have great potential for wood adhesive applications. Hemp proteins were extracted from two types of hempseed meals: Hlesia and commercial hempseed meals. Then the extracted hemp proteins with and without modification were used to prepare wood adhesives. Glutaraldehyde, previously identified as a toxic but effective crosslinker for soybean protein adhesives, was used as the crosslinking reagent for hemp protein in this study. Results showed that the wet adhesion strength of hemp proteins without modification was similar or superior compared to published data such as modified soy protein-based adhesives. Glutaraldehyde crosslinker only improved wet adhesion of commercial hemp protein at low press temperature such as 150 °C and 150–170 °C for Hlesia hemp protein. At press temperature 170–190 °C, wet adhesion of commercial hemp protein was not affected by glutaraldehyde crosslinker, while for Hlesia hemp protein, wet adhesion was not affected by glutaraldehyde crosslinker at 190 °C. The highly hydrophobic amino acid contents of hempseed proteins were identified to attribute to the unique wet adhesion performance for low water solubility and water resistance, which is essential for plant protein-based adhesive application. This is the first time to demonstrate that native hemp proteins could be a feasible and valuable alternative source of plant protein-based adhesives.
Plant proteins are attracting increased research attention, and the modification of their functional properties is a key area of current research. Industrial hemp is a new source of protein which can be produced in a sustainable manner. Hemp protein is rich in essential amino acids such as leucine, lysine, and phenylalanine, and also comes with good digestibility and various health benefits. Despite its potential, the scope of its application is limited due to its less-than-desirable technofunctional properties including solubility, emulsifying, encapsulating and gelling properties. This review provides a snapshot of various physicochemical and enzymatic methods that are currently used to enhance the aforementioned properties of plant proteins, particularly focusing on the modification of hemp protein. It compares the structural characteristics, physicochemical properties that can be positively affected by these methods and explains the underlying principles. This review highlights the fact that combination of two or more methods and particularly implementation of protein–polysaccharide complex coacervation and protein–polyphenol conjugation and peptide–polysaccharide conjugation greatly improve the technofunctional properties and help broaden the scope of application of hemp protein.
The scientific monograph presents theoretical and practical aspects of the development of science, technology, and innovation in the context of the global transformation of society. It covers general issues of technical, physical, and mathematical sciences, biological and agricultural sciences, sociological sciences, social communications, and historical sciences. The publication is intended for scientists, educators, postgraduate students and undergraduates, as well as the general readership.
More and more people in recent years have turned to oilseeds for their daily nutritional needs. Some of the most potent sources of nutrients and bioactive compounds can be found in the byproducts of whole seeds, oils, meal, and cake. Nutritionists and food manufacturers alike tout their benefits, so it’s no surprise that customers are showing a lot of enthusiasm for these novel pro-health additives. As a result, many scientists, activists, and government agencies are looking into new trends in the food science and industry to partially replace animal proteins with proteins of vegetable origin, which would not only improve the pro-health values of meat products but also correspond to the need to reduce meat production for ethical reasons and care for the environment. Oilseed proteins are a valuable functional component or alternative source of protein, particularly for the bakery and meat industries, due to the presence of biologically active proteins and peptides with antioxidant, antihypertensive, or neuroprotective properties. This book chapter provides a concise synopsis of the nutritional and pro-health aspects of selected oilseeds, focusing on their use in enhancing the qualities of food products, oilseed protein composition, proteins and peptides biological activity, and the possibility of allergenicity.
Hemp (Cannabis sativa L.) seeds contain a high concentration of proteins and biologically active compounds. The protein content is even higher in case of lipid part removal in oil production. The remaining part is considered a leftover, usually being used in animal feed. The aim of this study was to investigate the physicochemical composition of hemp seed cake flour, its nutritional quality and its impact on bread quality parameters. The properties of hemp seed cake flour were assessed in terms of protein quality, mineral composition, polyphenols and antioxidant activity. Hemp seed cake proved to be an important source of high-quality protein (31.62% d.m.) with the presence of eight essential amino acids. The biologically active potential of hemp seed cake has been demonstrated by the high content of polyphenols, especially those from the Cannabisin group. Hemp seed cake flour was incorporated in wheat flour at levels from 5 to 40% (w/w) to investigate its influence on bread quality parameters. The addition of hemp seed cake flour increased the total phenol content of bread, thus greatly enhancing the antioxidant activity. The protein content of bread was found to be enhanced from 11.11% d.m (control sample) to 18.18% d.m (for sample with 40% hemp seed cake flour). On the other hand, the addition of hemp seed cake flour led to decreased bread porosity, increased hardness and decreased resilience in the seed cake. Although, all bread samples recorded sensorial attributes ranging between “slightly like” and “like it very much”.
Even though plant proteins are more plentiful and affordable than animal proteins in comparison, direct usage of plant-based proteins (PBPs) is still limited because PBPs are fed to animals as feed to produce animal-based proteins. Thus, this work has comprehensively reviewed the effects of various factors such as pH, temperature, pressure, and ionic strength on PBP properties, as well as describes the protein interactions, and extraction methods to know the optimal conditions for preparing PBP-based products with high functional properties and health benefits. According to the cited studies in the current work, the environmental factors, particularly pH and ionic strength significantly affected on physicochemical and functional properties of PBPs, especially solubility was 76.0% to 83.9% at pH = 2, while at pH = 5.0 reduced from 5.3% to 9.6%, emulsifying ability was the lowest at pH = 5.8 and the highest at pH 8.0, and foaming capacity was lowest at pH 5.0 and the highest at pH = 7.0. Electrostatic interactions are the main way for protein interactions, which can be used to create protein/polysaccharide complexes for food industrial purposes. The extraction yield of proteins can be reached up to 86–95% with high functional properties using sustainable and efficient routes, including enzymatic, ultrasound-, microwave-, pulsed electric field-, and high-pressure-assisted extraction. Nondairy alternative products, especially yogurt, 3D food printing and meat analogs, synthesis of nanoparticles, and bioplastics and packaging films are the best available PBPs-based products. Moreover, PBPs particularly those that contain pigments and their products showed good bioactivities, especially antioxidants, antidiabetic, and antimicrobial.
Protein isolates of beach pea were prepared using sodium hydroxide (NaOH) and sodium hexametaphosphate (SHMP). Functional properties of the isolates so prepared were investigated and compared with those of other pea samples. Protein isolate of beach pea, prepared via NaOH extraction, had a protein content of 86.6%, while SHMP-extracted isolates contained 85.1% protein. Corresponding values for NaOH- and SHMP-extracted green pea and grass pea were 90.6, 89.9, 90.6 and 88.3%, respectively. Sulphur-containing amino acids were more prevalent in SHMP-extracted beach pea and green pea, while they were higher in NaOH-extracted grass pea. Tryptophan content was higher in NaOH-extracted than SHMP-extracted isolates. The predicted biological value and protein efficiency ratio of beach pea protein isolates indicated the high quality of products so prepared. Beach pea protein isolates exhibited a minimum solubility at pH 4.5. The pH and NaCl concentration effectively changed the functional properties of protein isolates. Beach pea protein isolates (NaOH- and SHMP-extracted) had in-vitro digestibility of 80.6 to 82.6% for pepsin-trypsin and 78.6 to 79.2% for pepsin-pancreatin.
The chapter discusses the protein stability with emphasis on compact globular proteins representing a single cooperative system. All the small compact globular proteins represent cooperative systems; they exhibit an extreme cooperativity that integrates the whole of their structure into a single structural unit. The large proteins, to which fibrillar proteins are also related, do not present single cooperative systems, but are subdivided into definite cooperative subsystems—structural blocks or domains. The advances in studying the stability of complicated proteins are connected with two methodical achievements: (1) the appearance of the precise scanning microcalorimetric technique, which affords reliable information on the heat capacity function of proteins in a broad temperature range; and (2) realization of the fact that the complicated heat effect of disruption of a complex macromolecular structure can be analyzed thermodynamically. The thermodynamic specificity of collagen has been considered. The volume of globular proteins does not increase at denaturation but decreases, as seen from their ability to denature under high pressure. The results of calorimetric studies are discussed, presenting the specific melting enthalpy of various protein structures—globular proteins, double-stranded coiled coils, and triplestranded coiled coils. The practical importance of thermodynamic studies of protein stability—that is, its importance not only for understanding the principles of organization of these molecules, but just for obtaining structural information on the domain level is emphasized.
Heat denaturation of several proteins from plant sources was studied using differential scanning calorimetry (DSC). Differences in temperatures of denaturation (Td), overall thermal transitions (ΔH), and half band widths (HBW) were noted for these proteins, with oats having the highest Td (112°C) and ΔH (4.45 cal·g−1) and the lowest HBW (9.2°C). Vital gluten, on the other hand, did not exhibit a characteristic thermogram so these parameters could not be measured. Using fababean protein as a model system, heat denaturation was shown to be irreversible. The effect of pH on the heat stability of the fababean protein indicated decreases in Td and ΔH on either side of the isoelectric point to a situation where the protein was believed to be completely denatured if the pH were taken below pH 2.5 or above 11.5. The irreversibility of this phenomenon in the alkaline region was demonstrated by DSC of alkaline extracted, isoelectrically precipitated fababean protein isolates.
Protein digestibility (in vitro) of dry seeds of moth bean cultivars varied significantly from 58.69 to 62.06%. The protein digestibility improved significantly when the seeds were given different domestic processing and cooking treatments including soaking, sprouting, ordinary cooking and pressure cooking. Pressure cooking of the seeds presoaked in plain water or mixed mineral salt solution had the most pronounced effect.
The effect of a dry heat treatment on trypsin inhibitors, protein quality and molecular weight of products obtained from enzymatic hydrolysis of chickpea flours was analysed in order to use chickpeas as food protein hydrolysates. Chickpea flour obtained from dehusked seeds was processed under different conditions, either by heating or enzymatic treatment. Heat treatment at 140 °C for varying times (1–24 h) inactivated trypsin inhibitors and facilitated enzymatic treatment but showed an unacceptable loss in the nutritional quality of the protein for heating times longer than 6 h. Enzymatic treatment with a commercial protease, Alcalase 0.6L, at pH 8 and 50 °C, increased the protein nutritional value of the chickpea by breaking the protein chains into shorter peptide chains more suitable to human nutrition.
Gelation of globular proteins on heating involves two separate stages. The first is partial unfolding (denaturation) of the native globular structure; the second is intermolecular aggregation. Denaturation involves dissociation of intramolecular bonds (non-covalent and, in some cases, disulfide) and is therefore an endothermic process. The aggregation step involves formation of new bonds between protein molecules, and would therefore be expected to give a differential scanning calorimetry (DSC) exotherm on heating, but numerous previous studies of the thermal gelation of whey proteins, carried out on conventional (fast scanning) DSC calorimeters (typical sample mass ∼15–50 mg), have shown only endothermic transitions. In the present work, however, we have observed the endothermic (denaturation) and exothermic (aggregation) processes in thermogelation of whey protein isolate (WPI) as separate transitions in DSC heating traces recorded on a Setaram microcalorimeter (sample mass ∼850 mg). Under conditions where aggregation occurs much more slowly than denaturation (low protein concentration; low ionic strength) the two transitions are well resolved, with the exotherm from aggregation following the endotherm from denaturation. The position of the exotherm, however, appears to be time-dependent rather than temperature-dependent. On reduction in heating rate, the apparent peak-maximum temperature of the aggregation exotherm decreases towards the (essentially constant) position of the denaturation endotherm, and, at sufficiently low scan rates, the exotherm becomes obscured by the more intense endotherm. Progressive displacement of the exotherm into the temperature range of the denaturation endotherm also occurs in response to changes that accelerate intermolecular aggregation and accompanying gelation (addition of salt; increasing protein concentration). The absence of a detectable exotherm in previous studies using conventional calorimeters is attributed to the much smaller sample mass than in the Setaram instrument, giving much faster heat transfer, which may cause the exothermic heat flow from the slow aggregation process to be swamped by the endothermic heat flow from the more rapid denaturation process.
Chromatography of edestin on DEAE-cellulose at 50° yields substantially one component. Edestin, however, has been fractionated into three components on DEAE-cellulose by a discontinuous temperature sodium chloride elution scheme. One of these components is a Lowry-positive, non-nitrogeneus material extracted from the seed and precipitated with the protein. The other two components are protein, and their amino acid compositions as well as their solubilities are distinct. The fact that twice-reprecipitated edistin is homogeneous with respect to ultracentrifugation but can be fractionated at 50° into distinct components gives strong support to the hypothesis that the material that has been classically regarded as edestin consists of several protein and non-protein components, among which there is a high degree of interaction. The solubility and chromatographic properties of crystalline edestin vary significantly with the previous history of the hempseed source. Edestin prepared from heat-treated, non-viable seed is insoluble at ordinary temperatures and has distinctly different elution characteristics.
The seed of Cannabis sativa L. has been an important source of nutrition for thousands of years in Old World cultures. Non-drug varieties of Cannabis, commonly referred to as hemp, have not been studied extensively for their nutritional potential in recent years, nor has hempseed been utilized to any great extent by the industrial processes and food markets that have developed during the 20th century. Technically a nut, hempseed typically contains over 30% oil and about 25% protein, with considerable amounts of dietary fiber, vitamins and minerals. Hempseed oil is over 80% in polyunsaturated fatty acids (PUFAs), and is an exceptionally rich source of the two essential fatty acids (EFAs) linoleic acid (18:2 omega-6) and alpha-linolenic acid (18:3 omega-3). The omega-6 to omega-3 ratio (n6/n3) in hempseed oil is normally between 2:1 and 3:1, which is considered to be optimal for human health. In addition, the biological metabolites of the two EFAs, gamma-linolenic acid (18:3 omega-6; GLA) and stearidonic acid (18:4 omega-3; SDA), are also present in hempseed oil. The two main proteins in hempseed are edestin and albumin. Both of these high-quality storage proteins are easily digested and contain nutritionally significant amounts of all essential amino acids. In addition, hempseed has exceptionally high levels of the amino acid arginine. Hempseed has been used to treat various disorders for thousands of years in traditional oriental medicine. Recent clinical trials have identified hempseed oil as a functional food, and animal feeding studies demonstrate the long-standing utility of hempseed as an important food resource.
Soybean (Glycine maxL. Merr., cv. Dare) protein subunits were separated by gradient gel electrophoresis and analyzed by two-dimensional densitometry
with computer-aided volume integration. Significant differences in the time required to achieve equilibrium staining with
Coomassie Blue were revealed among the various polypeptides. Bands corresponding to lipoxygenase reached staining equilibrium
in 2.7 h, whereas longer periods were required for polypeptides of β-conglycinin (5.5 to 6.7 h) and of glycinin (8.6 to 9.2
h). These differences among polypeptides could be attributed in part to changes in gradient concentration within the polyacrylamide
gel. Optimal staining intensity among all soluble proteins extracted from soybean seed was reached after staining for 8 h.
Shorter than optimal staining times lead to significant underestimation of parameters such as the percentage of β-conglycinin
and glycinin of total soluble protein.
Proteins of Phaseolus angularis, Phaseolus calcaratus and Dolichos lablab seeds (24.9 to 26.5% dry weight) were mainly contributed by their cotyledons which had a proportion of 82.9 to 90.8% by weight of the whole seeds. The levels of all the essential amino acids of the legume seeds (415 to 443 mg g−1 protein) were above the FAO/WHO, (1990) requirement with the essential amino acid scores of the methionine and cystine being 1.02 to 1.19. Appreciable amounts of oligosaccharides of the raffinose family were found mainly in the cotyledons (3.52 to 4.72% dry weight). The total dietary fibre of the legume seeds (13.5 to 19.3% dry weight) consisted predominantly of insoluble dietary fibre (IDF). The cotyledons contained 45.9 to 51.4% dry weight of starch but had low contents of cellulose and lignin. Hulls were highly lignified and consisted predominantly of IDF, with values ranging from 70.7 to 74.0% dry weight. The principal sugar residue analysis revealed that pectins were the major polysaccharide present in the SDF of both the cotyledons and the hulls. While xylans and cellulose were the major polysaccharides present in the IDF of the hulls, xyloglucans and arabinose-rich pectic substances were the principal non-starch polysaccharides in the IDF of the cotyledons.
The thermal properties of Phaseolus angularis (red bean) globulin were studied by differential scanning calorimetry under various medium conditions. Red bean globulin (RGB) was fractionated by ion-exchange chromatography into a major fraction, with SDS–PAGE pattern corresponding to the 7S vicilin, and two minor fractions, probably representing residual vicilin and the 11S legumin, respectively. The thermogram of RBG showed a major endothermic peak at 86.4°C and a minor transition at 92.2°C. Vicilin exhibited two endothermic peaks (87.7 and 94.1°C), while legumin showed one transition at 89.5°C. The progressive increase in denaturation temperature (Td) with increase in salt concentration, suggests a more compact conformation for RBG with higher thermal stability. Decreases in enthalpy and Td were observed under the influence of highly acidic and alkaline pHs, chaotropic salts, and protein perturbants such as sodium dodecyl sulfate, urea and ethylene glycol, indicating partial denaturation and decrease in thermal stability. Dithiothreitol and N-ethylmaleimide have little effect on the thermal properties of RBG since the major protein component, vicilin, is devoid of disulfide bonds.
Effects of limited enzymatic hydrolysis induced by trypsin on the physicochemical and functional properties of hemp (Cannabis sativa L.) protein isolate (HPI) were investigated. The enzymatic hydrolysis was confirmed by sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and size exclusion chromatography (SEC). SEC and differential scanning calorimetry (DSC) analyses confirmed the presence of aggregates in the corresponding hydrolysates (with the degree of hydrolysis of 2.3–6.7%). Functional properties, including protein solubility (PS), thermal properties, emulsifying and foaming properties, and water holding and fat adsorption capacities (WHC and FAC) were evaluated. The PS was remarkably improved by the limited enzymatic hydrolysis at all tested pH values. However, the enzymatic hydrolysis led to the marked decreases in emulsifying activity index, foaming capacity and foam stability, WHC and FAC. These decreases were to a great extent related to the presence of aggregates in the hydrolysates.
Using an improved method of gel electrophoresis, many hitherto unknown
proteins have been found in bacteriophage T4 and some of these have been
identified with specific gene products. Four major components of the
head are cleaved during the process of assembly, apparently after the
precursor proteins have assembled into some large intermediate
structure.
Edestin, a legumin class reserve protein from hemp seeds having six identical subunits was crystallized from ammonium phosphate at pH 5 and subsequently characterized by X-ray diffraction. The crystals are of space group R32 with a = 127 A and gamma = 116 degrees having an equivalent triply centered hexagonal cell of a = b = 215 A, c = 80 A. There is one hexameric protein in the rhombohedral unit cell, hence the subunits of the Edestin molecule must be arranged with 32 point group symmetry.
A 10-kDa protein was isolated from resting seeds of hemp (Cannabis sativa) by buffer extraction, gel filtration, ion-exchange chromatography, and reversed-phase high-pressure liquid chromatography. The protein did not inhibit bovine trypsin. It consisted of subunits composed of 27 and 61 residues and was held together by two disulfide bonds. The complete amino acid sequence was identified by protein analysis, and had 20 mole% of amino acids containing sulfur. The protein was most similar to a methionine-rich protein of Brazil nut (Bertholletia excelsa) and to Mabinlin IV, a sweetness-inducing protein of Capparis masaikai. The high methionine content and the absence of trypsin inhibitory activity suggested that the seed protein can be used to improve the nutritional quality of plant food-stuffs.
An in vitro protein digestion study, using pepsin, was carried out in uncooked and cooked sorghum and maize flour samples. The digestibility values from the uncooked samples showed that sorghum presents digestibility values similar to those of maize. In the case of the cooked samples, it was found that a wet cooking procedure promotes a decrease in sorghum protein digestibility when compared to maize. Electrophoresis was used to follow the in vitro pepsin sequential digestion procedure, and infrared spectroscopy was applied to establish its efficiency. SDS-PAGE results showed that both uncooked samples (sorghum and maize) behave in a similar way. The wet cooking procedure increases the amount of high molecular weight aggregates and promotes the appearance of two nonreducible and nondigestible 45 and 47 kDa proteins. These two protein fractions are directly related to the loss of digestibility. It was also shown that in cooked sorghum the monomers (gamma-, alpha-, and beta-) are more resistant to digestion than the corresponding uncooked samples.
The amino acid composition and physicochemical and functional properties of hemp (Cannabis sativa L.) protein isolate (HPI) were evaluated and compared with those of soy protein isolate (SPI). Edestin, a kind of hexameric legumin, was the major protein component. HPI had similar or higher levels of essential amino acids (except lysine), in comparison to those amino acids of SPI. The essential amino acids in HPI (except lysine and sulfur-containing amino acids) are sufficient for the FAO/WHO suggested requirements for 2-5 year old children. The protein solubility (PS) of HPI was lower than that of SPI at pH less than 8.0 but similar at above pH 8.0. HPI contained much higher free sulfhydryl (SH) content than SPI. Differential scanning calorimetry analysis showed that HPI had only one endothermic peak with denaturation temperature (T(d)) of about 95.0 degrees C, attributed to the edestin component. The T(d) of the endotherm was nearly unaffected by 20-40 mM sodium dodecyl sulfate but significantly decreased by 20 mM dithiothreitol (P < 0.05). The emulsifying activity index, emulsion stability index, and water-holding capacity of HPI were much lower than those of SPI, and the fat adsorption capacity was similar. The data suggest that HPI can be used as a valuable source of nutrition for infants and children but has poor functional properties when compared with SPI. The poor functional properties of HPI have been largely attributed to the formation of covalent disulfide bonds between individual proteins and subsequent aggregation at neutral or acidic pH, due to its high free sulfhydryl content from sulfur-containing amino acids.
The analysis of amino acid and elements in traditional chinese medicine cannabis. Plant Fiber and Products
- Y Yang
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Yang, Y., & Bai, W. (2001). The analysis of amino acid and elements in
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Protein quality evaluation
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