Dataset Information

OPTIMIZATION OF ALKALINE ?-AMYLASE PRODUCTION BY THERMOPHILIC BACILL US SUBTILIS.

ABSTRACT: Starch-degrading amylase enzyme is important in biotechnological applications as food, fermentation, textile, paper and pharmaceutical purposes. The aim of current study to isolate alkaline thermostable ?-amylase bacteria and then study the composition of medium and culture conditions to optimize cells growth and a-amylase production.Thermophilic amylase producing bacterium was isolated from local hot water-springs in Gazan city Saudi Arabia.Phylogenetic analysis of 16 S rRNA sequence for the strain revealed that the strain have the same sequence of Bacillus subtilis. Maximum amylase production was observed, when B. subtilis cultured in medium containing starch at concentration 0.5%, and 10 g/L peptones as nitrogen source at pH 8.5 in when it was incubated for 48 h at 45°C.An amylase-producing bacterium were isolated from hot-spring water and was identified as B. subtilis. Amylase produced from B.subtilis had optimum temperature 45°C and pH 8.5 in shaking media.

SUBMITTER: Al-Johani NB

PROVIDER: S-EPMC5411881 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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Publications

OPTIMIZATION OF ALKALINE Α-AMYLASE PRODUCTION BY THERMOPHILIC <i>BACILL US SUBTILIS</i>.

Al-Johani Nuha Bakeet NB Al-Seeni Madeha N MN Ahmed Youssri Mohamed YM

African journal of traditional, complementary, and alternative medicines : AJTCAM 20161123 1

<h4>Background</h4>Starch-degrading amylase enzyme is important in biotechnological applications as food, fermentation, textile, paper and pharmaceutical purposes. The aim of current study to isolate alkaline thermostable α-amylase bacteria and then study the composition of medium and culture conditions to optimize cells growth and a-amylase production.<h4>Materials and methods</h4>Thermophilic amylase producing bacterium was isolated from local hot water-springs in Gazan city Saudi Arabia.<h4>R ...[more]

PMID: 28480407

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Project description:BackgroundOur laboratory has constructed a Bacillus stearothermophilus α-amylase (AmyS) derivative with excellent enzymatic properties. Bacillus subtilis is generally regarded as safe and has excellent protein secretory capability, but heterologous extracellular production level of B. stearothermophilus α-amylase in B. subtilis is very low.ResultsIn this study, the extracellular production level of B. stearothermophilus α-amylase in B. subtilis was enhanced by signal peptide optimization, chaperone overexpression and α-amylase mutant selection. The α-amylase optimal signal peptide (SPYojL) was obtained by screening 173 B. subtilis signal peptides. Although the extracellular α-amylase activity that was produced by the resulting recombinant strain was 3.5-fold greater than that of the control, significant quantities of inclusion bodies were detected. Overexpressing intracellular molecular chaperones significantly reduced inclusion body formation and further increased α-amylase activity. Error-prone PCR produced an amylase mutant K82E/S405R (AmySA) with enzymatic activity superior to that of AmyS. Expression of the amySA gene with the SPYojL while overexpressing molecular chaperones resulted in a 7.1-fold improvement in α-amylase activity. When the final expression strain (WHS11YSA) was cultivated in a 3-L fermenter for 92 h, the α-amylase activity of the culture supernatant was 9201.1 U mL-1, which is the highest level that has been reported to date.ConclusionsThis is the first report that describes an improvement of B. stearothermophilus α-amylase extracellular production levels in B. subtilis using these strategies, and this represents the highest extracellular production level ever reported for α-amylase from B. stearothermophilus in B. subtilis. This high-level production provides a basis for enhanced industrial production of α-amylase. These extracellular production level improvement approaches are also expected to be valuable in the expression of other enzymes in B. subtilis.

Dataset Information

OPTIMIZATION OF ALKALINE ?-AMYLASE PRODUCTION BY THERMOPHILIC BACILL US SUBTILIS.

Publications

OPTIMIZATION OF ALKALINE Α-AMYLASE PRODUCTION BY THERMOPHILIC <i>BACILL US SUBTILIS</i>.

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