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Production of ?-Glucosidase from a Newly Isolated Aspergillus Species Using Response Surface Methodology.

ABSTRACT: A newly isolated fungus Aspergillus niger SOI017 was shown to be a good producer of ?-glucosidase from all isolated fungal strains. Fermentation condition (pH, cellobiose concentration, yeast extract concentration, and ammonium sulfate concentration) was optimized for producing the enzyme in shake flask cultures. Response surface methodology was used to investigate the effects of 4 fermentation parameters (yeast extract concentration, cellobiose concentration, ammonium sulfate concentration, and pH) on ?-glucosidase enzyme production. Production of ?-glucosidase was most sensitive to the culture medium, especially the nitrogen source yeast extract. The optimized medium for producing maximum ?-glucosidase specific activity consisted of 0.275% yeast extract, 1.125% cellobiose, and 2.6% ammonium sulfate at a pH value of 3.

SUBMITTER: Vaithanomsat P

PROVIDER: S-EPMC3118286 | biostudies-literature | 2011

REPOSITORIES: biostudies-literature

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Production of β-Glucosidase from a Newly Isolated Aspergillus Species Using Response Surface Methodology.

Vaithanomsat Pilanee P Songpim Molnapat M Malapant Taweesiri T Kosugi Akihiko A Thanapase Warunee W Mori Yutaka Y

International journal of microbiology 20110418

A newly isolated fungus Aspergillus niger SOI017 was shown to be a good producer of β-glucosidase from all isolated fungal strains. Fermentation condition (pH, cellobiose concentration, yeast extract concentration, and ammonium sulfate concentration) was optimized for producing the enzyme in shake flask cultures. Response surface methodology was used to investigate the effects of 4 fermentation parameters (yeast extract concentration, cellobiose concentration, ammonium sulfate concentration, and ...[more]

PMID: 21716658

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Project description:IntroductionA halophilic bacterium of the Halomonas elongata BK-AG25 has successfully produced ectoine with high productivity. To overcome the drawbacks of high levels of salt in the production process, a nonhalophilic bacteria of Escherichia coli (E. coli) was used to express the ectoine gene cluster of the halophilic bacteria, and the production of ectoine by the recombinant cell was optimized.MethodsThe ectoine gene cluster from the halophilic bacterium was isolated and inserted into an expression plasmid of pET30(a) and subsequently transformed into E. coli BL21 (DE3). Production of ectoine from the recombinant E. coli was investigated and then maximized by optimizing the level of nutrients in the medium, as well as the bioprocess conditions using response surface methodology. The experimental designs were performed using a central composite design.ResultsThe recombinant E. coli successfully expressed the ectoine gene cluster of Halomonas elongata BK-AG25 under the control of the T7 promoter. The recombinant cell was able to produce ectoine, of which most were excreted into the medium. The optimization of ectoine production with the response surface methodology showed that the level of salt in the medium, the incubation temperature, the optical density of the bacteria before induction, and the final concentration of the inducer gave a significant effect on ectoine production by the recombinant E. coli. Interestingly, the level of salt in the medium and the incubation temperature showed an inverse effect on the production of intracellular and extracellular ectoine by the recombinant cell. At the optimum conditions, the production yield was about 418 mg ectoine/g cdw (cell dry weight) after 12 hours of incubation.ConclusionThis study is the first report on the expression of an ectoine gene cluster of Halomonas elongata BK-AG25 in E. coli BL21, under the control of the T7 promoter. Optimization of the level of nutrients in the medium, as well as the bioprocess condition using response surface methodology, has successfully increased the production of ectoine by the recombinant bacteria.

Dataset Information

Production of ?-Glucosidase from a Newly Isolated Aspergillus Species Using Response Surface Methodology.

Publications

Production of β-Glucosidase from a Newly Isolated Aspergillus Species Using Response Surface Methodology.

Similar Datasets

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