Expression of α-Cyclodextrin Glycosyltransferase Gene of <i>Gebacillius</i> sp. CHB1 in <i>Bacillus subtilis</i>

CHEN Long-jun; LIN Chen-qiang; ZHANG Hui; JIA Xian-bo; FANG Yu; CHEN Ji-chen

doi:10.19303/j.issn.1008-0384.2019.05.014

Volume 34 Issue 5

Aug. 2019

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Article Navigation > Fujian Journal of Agricultural Sciences > 2019 > 34(5): 600-605

CHEN Long-jun, LIN Chen-qiang, ZHANG Hui, JIA Xian-bo, FANG Yu, CHEN Ji-chen. Expression of α-Cyclodextrin Glycosyltransferase Gene of Gebacillius sp. CHB1 in Bacillus subtilis[J]. Fujian Journal of Agricultural Sciences, 2019, 34(5): 600-605. doi: 10.19303/j.issn.1008-0384.2019.05.014

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CHEN Long-jun, LIN Chen-qiang, ZHANG Hui, JIA Xian-bo, FANG Yu, CHEN Ji-chen. Expression of α-Cyclodextrin Glycosyltransferase Gene of Gebacillius sp. CHB1 in Bacillus subtilis[J]. Fujian Journal of Agricultural Sciences, 2019, 34(5): 600-605. doi: 10.19303/j.issn.1008-0384.2019.05.014

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Expression of α-Cyclodextrin Glycosyltransferase Gene of Gebacillius sp. CHB1 in Bacillus subtilis

doi: 10.19303/j.issn.1008-0384.2019.05.014

Soil and Fertilizer Institute, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350003, China

Received Date: 2019-03-12
Rev Recd Date: 2019-04-22
Publish Date: 2019-05-28

Abstract

Abstract

Objective To construct a vector for expressing α-cyclodextrin glycosyltransferase (α-CGTase) gene in Bacillus subtilis. Method The α-CGTase gene from Gebacillius sp. CHB1 was amplified by PCR. The EcoR Ⅰ-digested pBES and Xho Ⅰ-digested α-CGT gene were connected and transformed into B. subtilis RIK1285. Subsequently, fermentation of the recombinant B. subtilis RIK1285/pBE-CGT was optimized. Result (1) The α-CGTase gene was expressed in a fermentation medium to show an enzymatic activity of 2.9 U·mL^-1 by B. subtilis RIK1285/pBE-CGT. (2) Medium TB with the formula of 0.5% glycerol, 1.2% peptone, 2.4% yeast extract, 1.64% K₂HPO₄, and 0.23% KH₂PO₄ was found to be optimal for the fermentation. After fermentation in TB at 37℃ for 24h, the α-CGTase activity reached 5.3 U·mL^-1, which was 8-fold of what the wild Gebacillius sp. CHB1 could generate. Conclusion The engineered B. subtilis RIK1285/pBE-CGT was successfully obtained and the fermentation process optimized.
- Engineered Bacillus subtilis,
- α-cyclodextrin glycosyltransferase,
- expression,
- optimization

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References(14)

References

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