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Four variable factors, their relative levels and assignment of experimental factors of the L9 Taguchi's array and relative enzyme activities.
Source publication
marcescens B4A was optimized following Taguchi’s
array methods. Twenty-three bacterial isolates were
screened from shrimp culture ponds in the South of
Iran. A chitinase-producing bacterium was isolated
based on it’s ability to utilize chitin as the sole carbon
source. The isolate designated as B4A, was identified
as Serratia marcescens based on it...
Contexts in source publication
Context 1
... optimization of medium constituents was carried out to improve chitinase activity of the selected strain. The identification of important factors was carried out in accordance with the enzymatic activity of the cells after 48 h of growth; thus 4 variables at 3 levels were investigated (Table 1). To examine the effects of these factors, Taguchi's arrays were used. ...
Context 2
... L9 array was selected to determine the effect of the four 3-level fac- tors on chitinase production. Based on the L9 Taguchi's design, 9 experiments were carried out in triplicate (Table 1). In the full-factorial experimental design procedures, at least 64 experiments are neces- sary to reach the same conclusions as those of the Taguchi's array method using this variety of factors (Ross, 1998;Roy, 1990). ...
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Citations
... However, the antagonistic effect of other microorganisms, such as strains of the genus Serratia, on Colletotrichum spp. or A. alternata has not been reported. There are some reports of the inhibitory effect of Serratia on other fungal genera [10,11]. It is possible that strains of the genus Serratia could be used for biological control of Colletotrichum spp. ...
... Colloidal chitin was prepared as reported by Zarei et al. [11]. Briefly, 24 g chitin powder obtained from shrimp shell (C7170, Sigma-Aldrich Co., St. Louis, MO, USA) was added to 790 mL concentrated HCl and left at 4 • C overnight with vigorous stirring. ...
... The cultures were washed twice in sterile isotonic saline solution (ISS; 0.85% of NaCl) by centrifuging at 3500× g for 20 min and resuspending the pellets in ISS at about 10 9 CFU/mL. However, according to Zarei et al. [11], a minimum culture (MC) medium containing colloidal chitin was prepared with the following ingredients and proportions: 0.5% colloidal chitin, 0.03% peptone, 0.03% yeast extract, 0.07% K 2 HPO 4 , 0.03% KH 2 PO 4 , 0.05% MgSO 4 .7H 2 O, 1.5% agar, 0.2% NH 4 NO 3 , 0.1% NaCl (w/v), and 0.1% (v/v) trace elements (pH 7). The culture medium was sterilised by autoclaving and poured into Petri dishes. ...
Microorganisms represent a viable option for the control of phytopathogens. From the surface of healthy mangoes, different bacteria were isolated. For all isolated bacterial strains, we determined their antimicrobial activity against a fungal strain that caused anthracnose in mangoes and against Alternaria alternata, both in the culture medium and directly on mangoes. The bacterial strains with the highest antifungal activity were identified by sequencing the 16s rRNA gene. Two species of Serratia were identified: marcescens and nematodiphila. Finally, the chitinolytic, glucanolytic, and cellulolytic activity and prodigiosin production of bacteria with antifungal activity was determined. Five fungal strains were isolated from mangoes with anthracnose. Only one strain was responsible for anthracnose in mangoes. This fungal strain was identified as Colletotrichum siamense. Against C. siamense and A. alternata in vitro and in mango selected strains of Serratia showed antifungal activity. Finally, the Serratia strains produced chitinases, glucanases, cellulases and prodigiosin, and the two S. marcescens strains did not produce hemolysins. The three Serratia strains isolated in this study can potentially be used in the biological control of anthracnose caused by C. siamense and A. alternata on mango.
... Prodigiosin is a red microbial pigment with a tripyrrole ring structure [1][2][3]. It is mainly produced by Serratia marcescens [4,5], Pseudomonas, Actinomycetes, and some marine bacteria [6][7][8]. Prodigiosin appears red under acidic and neutral conditions and yellow under alkaline conditions [9,10]. ...
Prodigiosin is a red pigment produced by Serratia marcescens with anticancer, antimalarial, and antibacterial effects. In this study, we extracted and identified a red pigment from a culture of S. marcescens strain ZPG19 and investigated its effect on the growth performance and intestinal microbiota of Kunming mice. High-performance liquid chromatography/mass spectrometry revealed that the pigment had a mass-to-charge ratio (m/z) of 324.2160, and thus it was identified as prodigiosin. To investigate the effect of prodigiosin on the intestinal microbiota, mice (n = 5) were administered 150 μg/kg/d prodigiosin (crude extract, 95% purity) via the drinking water for 18 days. Administration of prodigiosin did not cause toxicity in mice. High-throughput sequencing analysis revealed that prodigiosin altered the cecum microbiota abundance and diversity; the relative abundance of Desulfovibrio significantly decreased, whereas Lactobacillus reuteri significantly increased. This finding indicates that oral administration of prodigiosin has a beneficial effect on the intestinal microbiota of mice. As prodigiosin is non-toxic to mouse internal organs and improves the mouse intestinal microbiota, we suggest that it is a promising candidate drug to treat intestinal inflammation.
... NRG4 were 1.41 mg mL -1 and 74.07 µM (µg h) -1 , respectively [9]. Km and Vmax chitinase from Serratia marcescens B4A was 8.3 mg mL -1 and 2.4 mmol min -1 [10], Km and Vmax chitinase from B. cereus 11 UJ were 29.71 µg mL -1 and 1.035 × 10 -1 µg (mL s) -1 , respectively [11]. The lower the Km, the higher the enzyme activity towards colloidal chitin as substrat. ...
Chitinases is an enzyme capable of degrading chitin into oligomers to produce chitin derivatives products which are more useful. Thermostable-chitinase is of important in the relevant industrial application, since the degradation process oftently requires prety high temperature. This research report a characterization of chitinase isolated from thermophilic microorganism. The chitinase was obtained from Bacillus licheniformis B2 isolated from Ijen hot spring, East Java. It has the best chitinolytic activity at pH 7 when colloidal chitin was used as substrate. The enzyme exhibited activity in broad temperature range, from 50 °C to 70 °C, optimally at 55 °C. It was stable at 50 °C up to 90 min, at 60 °C up to 60 min and at 70 °C up to 30 min. At neutral pH this enzyme has negative charge but further purification is needed to determine its pI. The K m and V max of this chitinase for colloidal chitin were 101.96 mg mL ⁻¹ and 2.72 μmol (min mL) ⁻¹ , respectively. Addition of NaCl, KNO 3 and MgSO 4 decreased the activity of chitinase following mixed inhibitor mode. This enzyme should be a good candidate for applications in the recycling of chitin waste.
... In addition, microbial isolates were screened on agar plates containing 0.5% colloidal chitin (prepared according to [11]), 0.8% Nutrient Broth, 1% malt extract, 1% peptone, 0.1% NaCl and 2% bacteriological agar (w/v) [12] to observe chitinolytic activity (CHIT). Chitinase quantification was assayed by measuring the release of N-acetyl-D-glucosamine (NAGA) from colloidal chitin [13]. ...
Strains isolated during composting processes of plant waste, and identified as Actinobacteria, proved to be significant producers of compounds that actively participate in the control of phytopathogens, such as those that cause Damping-off disease. Although most of the actinomycetes analyzed showed to be antagonistic strains against common phytopathogens, only some 30% proved to be capable of producing bioactive substances, such as siderophores, salicylic acid, chitinase enzymes or cyanide, so that antibiosis could be considered the most probable antagonistic mechanism for a high proportion of the strains investigated. 6% of the microorganisms identified in this work, were selected as potential strains to be investigated in depth, since they further stimulated plant growth (germination index tests greater than 100%). Microbacteriaceae was one of the most prominent families.
... This could be attributed to the presence of nitrogenous compounds, growth factors, and oligomers of GlcNAc in this medium and its addition had a stimulatory effect on cell growth (Nawani and Kapadnis, 2005). Zarei et al. (2010) reported that malt extract was the optimum nitrogen source for chitinase production by S. marcescens B4A. ...
Thirty-four bacterial isolates were isolated from soil samples collected from the North Western Coast and a water sample collected from brackish water at Siwa Oasis, Matrouh Governorate, Egypt. Only six isolates showed chitinase activity when screened on colloidal chitin agar medium. The highest chitinolytic activity was achieved by a bacterial isolate labeled as A.S. This isolate was identified as Aeromonas hydrophila based on analysis of 16S rRNA gene sequence and morphological, physiological, and biochemical characteristics. Optimization of the cultural conditions for maximum chitinase production by A. hydrophila revealed that the highest level of chitinase was recorded when the bacterium was grown in malt nitrogen-based medium containing 1% colloidal chitin at pH7 for 48 h incubation at 30 °C. Crude chitinase from isolate A. hydrophila was evaluated against first instar larvae of the greater wax moth; Galleria mellonella L. (Lepidoptera: Pyralidae) at different concentrations of 0, 185, 205, 235, 265, 295 U/mg protein. It increased larval and pupal mortality rates in a concentration-dependent manner. The tested crude chitinase significantly induced a decrease in adults’ emergence rate and their fecundity.
... The optimum incubation period for optimum chitinase production by Serratia marcescens was after 6 days of incubation [35], which in accordance to the result from this research. This result is also supported by a previous research which stated that the optimum activity and the optimum amount of chitinase enzyme produced by S. marcescens were produced at pH 7.9 [39]. ...
Objective: The aim of this research was to determine the optimum condition for Serratia marcescens to produce optimum amount of N-acetyl glucosamine using chitin isolated from tiger shrimp (Penaeus monodon) shells.Methods: This research was conducted using submerged fermentation method. The treatments used were various fermentation temperatures (20, 30, and 37°C), pH (6, 7, and 8), and incubation period (2, 4, 6, and 8 days).Results: Chitinolytic index of Serratia marcescens was 2.203±0.59 after 2 days of incubation. Optimum temperature for N-acetyl glucosamine production using S. marcescens was at 30°C. Optimum pH and incubation period for N-acetyl glucosamine production were at pH 8 and 6 days of incubation period.Conclusion: S. marcescens is able to ferment chitin from shrimp shell to produce N-acetyl glucosamine of 41,166.11±4,480.59 mg/l at optimum fermentation condition.
... In the current study, the production of the chitinolytic enzyme from B. atrophaeus A7 was observed since the first day following the bacterial cultivation under experimental conditions; however, the decrease in activity with a prolonged growth regime after the 8 th day might be due to the reduced level of nutrients in the culture medium and/or denaturation of chitinase by proteases (Zarei et al., 2010). The optimum enzyme activity and stability of the isolated chitinase from B. atrophaeus A7 at different temperatures and pH values coincided with previously reported bacterial chitinases. ...
Microbial chitinases are important environmental biomolecules with biotechnological and medicinal applications in addition to being a source of environmental friendly biopesticides. They are considered as safe alternatives to some available chemical insecticides, especially against insects that may act as an intermediate hosts as well as vectors between manifested plant materials and humans. A crude chitinolytic enzyme was isolated from isolate A7 (Bacillus atrophaeus Nakamura 1989). The isolate was identified based on the morphological and biochemical characteristics as well as the sequencing of 16S rRNA. The produced enzyme had a total activity of 68.9±1.03 mU/mL; a specific activity of 2670±40.2 mU/mg protein, and was optimally active at 40°C, 4-9 pH with stability for one hour at 30-40°C and 6-7 pH. It was inhibited by Cu²⁺, Fe³⁺, Ni³⁺, Zn²⁺ and Ba²⁺ metal ions and impeded the development of 50 % of Drosophila melanogaster larvae into adults (LD50) at 17.3±1.4 mU/mL. In this study, the larvicidal activity of chitinase from B. atrophaeus is explored for the first time with the potential of being applied as environmental friendly biopesticide technologies.
... Using this approach, we apply a system of arrays which permits estimation of a maximum number of key effects in an orthogonal fashion with a minimum number of experimental runs [14]. This method was applied in different fields of biotechnology such as food and industrial fermentations [15][16][17], Molecular biology [18][19][20], wastewater treatment and bioremediation [16,21,22] and health care [23][24][25]. In this work, we explained the application of this method for optimization of medium culture parameters, for producing FSH, a complex glycoprotein hormone, in the recombinant CHO-c111 cell line. ...
... The micrograph suggests that it is a motile, rod-shaped bacterium with rounded ends, which are the most common characteristic features of S. marcescens. Zarei et al. (2010) and Khanam and Chandra (2015) also reported the same morphological features for S. marcescens B4A and S. marcescens KC1. To characterize and identify the bacterial isolate S. marcescens ABHI001 that exhibited the maximum degradation percentage of p-cresol, both its biochemical and physiological characteristics were examined. ...
... Furthermore, biochemical tests, such as urease, catalase, oxidase, and indole production; methyl-red; Voges-Proskauer; and citrate (IMViC) utilization tests were performed, and the Table 1. Zarei et al. (2010) performed biochemical and microbiological analyses to characterize the newly isolated S. marcescens B4A and found that this isolate was indole negative, methyl-red negative, and citrate positive. They also performed the Voges-Proskauer test and found red color formation, indicating its positive reaction. ...
This study evaluated the capability of Serratia marcescens ABHI001 to effectively degrade p-cresol through different techniques. The molecular identity of the laboratory isolate S. marcescens ABHI001 was confirmed through the 16S ribosomal DNA gene pattern, and its morphological features were investigated through field-emission scanning electron microscopy. In addition, the degradation behavior of the isolate for cresol was verified using several techniques, including UV–visible spectroscopy, followed by high-performance liquid chromatography (HPLC), gas chromatography, and Fourier transform infrared spectroscopy. The maximum degradation percentage of 85% for p-cresol could be achieved after 18 h of incubation with S. marcescens ABHI001. The formation of p-hydroxybenzaldehyde, p-hydroxybenzoate, and protocatechuate metabolites was confirmed through HPLC. The study results indicate that S. marcescens ABHI001 may have applications in the bioremediation of organic pollutants, such as p-cresol.
... It has been found that waste generated on the processing of shrimps and crabs contains up to 50% of chitin by dry weight [4,5]. Chitin, a highly insoluble biopolymer, is one of the most abundant organic compounds in nature (after cellu-most of these organisms have been reported to utilize pure chitin as a carbon source [15,16]. Some organisms have been reported to produce chitinase [17,18] and chitin oligosaccharides [19,20,10] utilizing seafood waste. ...
Disposal of chitinaceous waste is a major problem of seafood industry. Most of the known chitinolytic organisms have been studied with respect to pure chitin as substrate. Use of these organisms for degradation of seafood waste has not been explored much. In present study a marine bacterium capable of proficiently degrading shrimp waste with co-production of value added products like chitinase and chitin oligosaccharides was isolated from seafood waste dumping sites. On 16s rRNA and biochemical analysis bacterium was found to be a novel species of genus Paenibacillus.Under optimized condition complete shrimp waste degradation (99%) was achieved along with chitinase yield of 20.01IUml-1. SEM and FTIR showed the structural changes and breakage of bonds typical to that of chitin, which indicated that this process can be used for the degradation of other chitinaceous material also. Thin layer chromatography revealed the presence of chitin oligosaccharides of various degree of polymerization in the hydrolysate. Complete degradation of shrimp waste by Paenibacillus sp. AD makes it a potential candidate for the bioremediation of seafood waste at large scale. Concomitant production of chitinase and chitin oligosaccharides further makes the process economical and commercially viable.
