Complete secretion of recombinant Bacillus subtilis levansucrase in Pichia pastoris for production of high molecular weight levan
文献类型： 外文期刊
作者： Chen, Shuochang 1 ; Tong, Qiuping 1 ; Guo, Xiaolei 1 ; Cong, Hao 1 ; Zhao, Zi 1 ; Liang, Wenfeng 1 ; Li, Jiemin 4 ; Zhu, Ping 1 ; Yang, Hui 1 ;
作者机构： 1.Guangxi Univ, Coll Life Sci & Technol, 100 Daxue Rd, Nanning 530004, Guangxi, Peoples R China
2.Guangxi Univ, State Key Lab Conservat & Utilizat Subtrop Agrobio, 100 Daxue Rd, Nanning 530004, Guangxi, Peoples R China
3.Guangxi Res Ctr Microbial & Enzyme Engn Technol, 100 Daxue Rd, Nanning 530004, Guangxi, Peoples R China
4.Guangxi Acad Agr Sci, Agrofood Sci & Technol Res Inst, 174 East Daxue Rd, Nanning 530007, Guangxi, Peoples R China
关键词： Thermostable levansucrase; Pichia pastoris; Complete secretion; Glycosylation; High molecular weight levan
期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：8.025； 五年影响因子：7.626 ）
ISSN： 0141-8130
年卷期： 2022 年 214 卷
页码：
收录情况： SCI
摘要： Three signal peptides from alpha-mating factor (alpha-MF), inulinase (INU) and native levansucrase (LS) were compared for secretion efficiency of Bacillus subtilis levansucrase SacB-T305A in Pichia pastoris GS115. The first complete secretion of bacterial levansucrase in yeasts under methanol induction was achieved while using alpha-MF signal. The secreted recombinant Lev(alpha-MF) proved to be glycosylated by combination of NanoLC-MS/MS and Endo H digestion. Interestingly, glycosylation not only improved significantly the polymerase thermostability, but also reversed the products profiles to favor synthesis of high molecular weight (HMW) levan which accounted for approximately 73 % to total levan-type polysaccharides. It indicated for the first time that the glycosylation of recombinant B. subtilis levansucrase affected significantly the products molecular weight distribution. It also provided a promising enzymatic way to effectively product HMW levan from sucrose resources.
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