期刊文献+

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 被引量:2

Construction of Lower Optimum pH Mutants of Bacillus flexus β-Amylase and Its Application in Maltose Production
在线阅读 下载PDF
导出
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。 Beta-amylase and other starch hydrolases were complexed to produce high purity maltose syrup in industry.Compared with plantβ-amylase,the microbialβ-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 microbialβ-amylase is 6.0~8.0,which is difficult to complex with other starch hydrolases.In the pvious study,the optimum pH of Bacillus flexusβ-amylase was 7.0.In the present study,three B.flexusβ-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 L396Kβ-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.
作者 亓旭辉 吴敬 王蕾 陈晟 QI Xuhui;WU Jing;WANG Lei;CHEN Sheng(State Key Laboratory of Food Science and Technology,Jiangnan University,Wuxi 214122,China;Key Laboratory of Industrial Biotechnology,Ministry of Education,Jiangnan University,Wuxi 214122,China)
出处 《食品与生物技术学报》 CAS CSCD 北大核心 2019年第10期105-110,共6页 Journal of Food Science and Biotechnology
基金 国家杰出青年科学基金项目(31425020) 江苏省产学研前瞻性联合研究项目(BY2015019-32) 江苏高校优秀科技创新团队项目(2015) 111计划(111-2-06)
关键词 Β-淀粉酶 定点突变 麦芽糖 弯曲芽孢杆菌 酶转化 β-amylase site-directed mutagenesis maltose B.flexus enzyme conversion
  • 相关文献

参考文献1

二级参考文献17

  • 1Duan X,Chen J,Wu J. Optimization of pullulanase production in Escherichia coli by regulation of process conditions and supplement with natural osmolytes[J]. Bioresource Technology, 2013,146: 379-385.
  • 2Ali G,Rihouey C,Le Cerf D,et al. Effect of carboxymethyl groups on degradation of modified pullulan by pullulanase from Klebsiella pneumoniae[J]. Carbohydrate Polymers ,2013,93 ( 1 ) : 109-115.
  • 3Kunamneni A,Singh S. hnproved high thermal stability of pullulanase from a newly isolated thermophilic Bacillus sp. AN-7[J]. Enzyme and Microbial Technology, 2006,39 (7) : 1399-1404.
  • 4Shi M,Chen Y,Yu S,et al. Preparation and properties of RS III from waxy maize starch with pullulanase [J]. Food Hydrocolloids, 2013,33 (1) : 19-25.
  • 5Doman-Pytka M, Bardowski J. Pullulan degrading enzymes of bacterial origin[J]. Crit Rev Microbiol, 2004,30 (2) : 107-121.
  • 6吉本斯布坎南.伯杰细菌鉴定手册[Z].第8版.北京:科学出版社,2008.
  • 7Wu K M, Li L H ,Yan J J ,et al. Genome sequencing and comparative analysis of Klebsiella pneumoniae NTUH-K2044 ,a strain causing liver abscess and meningitis[J]. J Baeteriol,2009,191 (14):4492-4501.
  • 8Fouts D E,Tyler H L,DeBoy R T,et al. Complete genome sequence of the N2-fixing broad host range endophyte Klebsiella p neumoniae 342 and virulence predictions verified in mice[J]. PLoS Genet,2008,4(7 ):el 000141.
  • 9Nair S U,Singhal R S,Kamat M Y. Induction of pullulanase production in Bacillus cereus FDTA-13 [J]. Bioresource Technology, 2007,98 (4) : 856-859.
  • 10李杨霞,张心齐,林盈盈,王勇,吴敏.我国中部地区热泉嗜热菌的分离及其产酶研究[J].浙江大学学报(理学版),2008,35(2):204-209. 被引量:10

共引文献1

同被引文献20

引证文献2

二级引证文献1

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部