Bacillus flexus β-淀粉酶低pH值突变体的构建及在麦芽糖制备中的应用
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Q814.9

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Construction of Lower Optimum pH Mutants of Bacillus flexus β-Amylase and Its Application in Maltose Production
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    摘要:

    工业上将β-淀粉酶与其他淀粉水解酶进行复配可获得高纯度的麦芽糖浆。相对于目前常用的植物来源β-淀粉酶,微生物β-淀粉酶具有生产工艺简单、不受原料限制、质量稳定、纯度高等优势,但其最适作用pH多为6.0~8.0,难以与其他淀粉水解酶(pH多为4.5~5.5)进行复配。本实验室前期获得的弯曲芽孢杆菌β-淀粉酶其最适pH值为7.0,本研究在此β-淀粉酶基础上构建T47K,Y164K,L396K三个突变体,其最适pH由突变前的7.0分别下降为6.0,4.5,5.5,其中L396K的最适温度也由突变前的50 ℃提高为60 ℃。以马铃薯淀粉(10% w/v)为底物进一步对L396K制备麦芽糖的转化条件进行探索与优化。结果表明,在反应初始pH为 5.5,温度60 ℃,β-淀粉酶加酶量为100 U/g干淀粉时麦芽糖最大转化率达到80.2%,符合高纯度麦芽糖浆的生产要求。

    Abstract:

    Beta-amylase and other starch hydrolases were complexed to produce high purity maltose syrup in industry. Compared with plant β-amylase,the microbial β-amylase has the advantages of high purity,little limitation with the raw materials,and the suitability for large-scale production. However,the optimum pH of the microbial β-amylase is 6.0~8.0,which is difficult to complex with other starch hydrolases. In the pvious study,the optimum pH of Bacillus flexus β-amylase was 7.0. In the present study,three B.flexus β-amylse mutants T47K,Y164K,L396K were constructed and the optimal pH were changed from 7.0 to 6.0,4.5 and 5.5 respectively. At the same time,the optimum temperature of L396K was increased from 50 ℃ to 60 ℃. The production conditions of maltose using L396K β-amylase was also optimized. When using potato starch(10% w/v) as the substrate under the condition of pH 5.5 and temperature 60 ℃,the yield of maltose reached 80.2%,which satisfied the production requirement of high purity maltose syrup.

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亓旭辉,吴敬,王蕾,陈晟.Bacillus flexus β-淀粉酶低pH值突变体的构建及在麦芽糖制备中的应用[J].食品与生物技术学报,2019,38(10):105-110.

QI Xuhui, WU Jing, WANG Lei, CHEN Sheng. Construction of Lower Optimum pH Mutants of Bacillus flexus β-Amylase and Its Application in Maltose Production[J]. Journal of Food Science and Biotechnology,2019,38(10):105-110.

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  • 在线发布日期: 2020-04-02
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