Construction and Fermentation Optimization of a Recombinant Bacillus subtilis Producing Alkaline Amylase

doi:10.3969/j.issn.1673-1689.2016.03.012

Home > Archive>Volume 35, Issue 3, 2016 >296-302. DOI:10.3969/j.issn.1673-1689.2016.03.012

Construction and Fermentation Optimization of a Recombinant Bacillus subtilis Producing Alkaline Amylase
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                        10.3969/j.issn.1673-1689.2016.03.012
                    
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Alkaline amylase，which could efficiently hydrolyze starch under alkaline conditions，has been widely used in textile desizing field，detergent industry，and pharmaceutical production. In this study，alkaline amylase from Alkalimonas amylolytica （AmyK） was screened through signal peptide and expressed in Bacillus subtilis WB600. The fermentation condition of the recombinant strain was then optimized. The extracellular AmyK activity of the recombinant B. subtilis could reach a maximal level of 146.86 U/mL with 51 h fermentation mediated by ywbN signal peptide. There was a protein band observed in the culture supernatant which was consistent for the theoretical molecular weight of 60 kDa for AmyK. The fermentation medium was optimize via a single factor test and determined as dextrin of 32 g/L，tryptone of 12 g/L，yeast extract of 24 g/L，KH2PO4 of 2.32 g/L，and K2HPO4·3H2O of 16.43 g/L with pH regulated by 0.2% w/v Na2CO3 after 24 h fermentation. The AmyK activity reached to 640.33 U/mL after 72 h，increasing by 336% when compared with the initial medium，which was the highest yield of alkaline amylase for the reported recombinant B. subtilis strains. This study provided a experimental support for the commercial production of AmyK from fermentation.

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WANG Jingli, LIU Song, DU Guocheng, CHEN Jian. Construction and Fermentation Optimization of a Recombinant Bacillus subtilis Producing Alkaline Amylase[J]. Journal of Food Science and Biotechnology,2016,35(3):296-302.

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