The Use of Crude Shrimp Shell Powder for Chitinase Production by Serratia marcescens WF
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Chitinase
Ultraviolet light and nitrosoguanidine were used to mutagenize a red pigmented culture of Serratia marcescens, strain EB415, which produced chitinase. After mutagenesis, a stable, non-pigmented mutant designated BL40 was isolated which produced larger colonies and zones of clearing on solid medium containing colloidal chitin. In liquid medium with colloidal chitin as the sole carbon source both strains grew similarly but BL40 produced 160 units/ml of chitinase compared with 60 units/ml for EB 415, an increase of 167%. When chitin concentration was increased in the medium, chitinase production also increased. Chitinase appeared to be extracellular, since the supernatant from washed, sonicated cells for both strains showed no detectable amount of chitinolytic activity.
Chitinase
Serratia
Carbon source
Ultraviolet light
Liquid medium
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Serratia marcescens strain was isolated at the laboratory from decomposed stalk of paddy straw mushroom (Volvariella volvacea) by using swollen chitin as a source of carbon. Morphological and biochemical characteristics of the bacterium were studied. The bacterium showed exponential growth up to 18 h after inoculation in batch culture. The maximum enzyme production by the bacterium was analyzed at 92 h of inoculation at 30°C. Similarly, with respect to different concentration of chitin, the minimal medium supplemented with 1.75% of swollen chitin produced maximum amount of chitinase enzyme. Therefore, the present study showed that isolated bacterium is a good source of chitinase. Moreover, though the bacterium was grown at the cheaper medium, the enzyme can be used for biodegradation of chitinous wastes as well as biological control of fungal pathogen at cheaper cost. Key words: Chitin, chitinase, swollen chitin, Serratia marcescens.
Chitinase
Volvariella volvacea
Serratia
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Swollen chitin, flake chitin, powder chitin, and mushroom paste were used as substrates for chitinase production by Serratia marcescens GG5 in submerged fermentation. Enzyme production was 0.3 U/ml when the organism was grown in M9 medium supplemented with 0.5% swollen chitin and 0.5% soluble starch. Scanning electron microscopy revealed that Serratia marcescens GG5 digested the chitin flakes by producing chitinase.
Chitinase
Serratia
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Chitinase
Marine bacteriophage
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Chitin from shrimp shells was prepared to evaluate the production of extracellular chitinase from Serratia marcescens QM B1466 in submerged culture under controlled conditions of temperature (28°C). shaking (150 rpm) and pH (7.7).To determine the composition of chitin on the shell, the material was ground, classified by sieving, deproteinized and demineralized. The proximate waste analysis indicates a high content of crude protein (36.11%). The highest chitinase activity during the enzymatic hydrolysis process was 1.91 µmols. equivalent to N-actetylglucosamine produced per mi of filtered culture: this determines the enzyme capacity of hydrolysis on the crystalline chitin. The percentage of chitin conversion during fermentation (73%) is the result of this activity. The N-acetylglucosamine produced is used by the bacterium for growth. which was adjusted to the Monod model.
Chitinase
Enzymatic Hydrolysis
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A sustainable, economical and convenient one-step process to produce high-value chitin oligomers and digestible shell residue from shrimp shell waste by a chitinase fused to a carbohydrate-binding module.
Chitinase
Carbohydrate-binding module
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Ultraviolet light and nitrosoguanidine were used to mutagenize a red pigmented culture of Serratia marcescens, strain EB415, which produced chitinase. After mutagenesis, a stable, non-pigmented mutant designated BL40 was isolated which produced larger colonies and zones of clearing on solid medium containing colloidal chitin.
In liquid medium with colloidal chitin as the sole carbon source both strains grew similarly but BL40 produced 160 units/ml of chitinase compared with 60 units/ml for EB 415, an increase of 167%. When chitin concentration was increased in the medium, chitinase production also increased. Chitinase appeared to be extracellular, since the supernatant from washed, sonicated cells for both strains showed no detectable amount of chitinolytic activity.
Chitinase
Serratia
Carbon source
Ultraviolet light
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Chitinase production by newly isolated Serratia 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 its 16S rRNA sequence and key morphological, physiological and biochemical characteristics. The cultivation of Serratia marcescens B4A in the appropriate liquid medium resulted in production of high levels of chitinase. The malt extract and colloidal chitin represented the best nitrogen and carbon sources, respectively. Chitinase production by Serratia marcescens B4A was optimized following the Taguchi orthogonal array (OA) for the design of experiments (DOE). Statistical experimental design via the Taguchi method was applied to determine the optimal levels of physical parameters and key media components in the medium, such as temperature, pH, NaCl and chitin concentrations. The results of this study showed that temperature of 30oC, pH 7.9, NaCl 0.1% (w/v) and chitin 1% (w/v) are optimal conditions for this protocol.
Chitinase
Serratia
Carbon source
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Chitinases play an important role in the decomposition of chitin and results in the utilization of chitin as a renewable resource. A total of 18 chitinolytic bacteria were isolated from the sand sample. Following primary and secondary screening in colloidal chitin medium, strain CBC-5 demonstrated the highest chitinolytic activity and was selected for further study. It was later identified as Serratia marcescens. Addition of easily metabolized sugars had inhibitory effect on the enzyme production and the highest yield was obtained with colloidal chitin as the sole source of carbon and yeast extract (1% w/w) as nitrogen source. 10.3 % increase in chitinase yield was observed when Triton X-100 was added to the medium. The optimum pH, temperature, and incubation period for chitinase production by the isolate was found to be 7.0, 30°C and 72 h respectively. Sodium dodecyl sulphate polyacrylamide gel electrophoresis revealed the apparent molecular weight of the enzyme to be 42 kDa. The results obtained in the present study showed that isolated bacterium is a potent producer of chitinase and the enzyme can be exploited for the biodegradation of chitinous wastes and may find applications as biocontrol agents against fungi and insects.
Chitinase
Yeast extract
Serratia
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