Complete secretion of recombinant Bacillus subtilis levansucrase in Pichia pastoris for production of high molecular weight levan

被引：8

作者：

Chen, Shuochang ^{[1
,2
,3
]}

Tong, Qiuping ^{[1
]}

Guo, Xiaolei ^{[1
,2
,3
]}

Cong, Hao ^{[1
]}

Zhao, Zi ^{[1
,2
,3
]}

Liang, Wenfeng ^{[1
,2
,3
]}

Li, Jiemin ^{[4
]}

Zhu, Ping ^{[1
]}

Yang, Hui ^{[1
,2
,3
]}

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

来源：

INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES | 2022年 / 214卷

关键词：

Thermostable levansucrase; Pichia pastoris; Complete secretion; Glycosylation; High molecular weight levan; GLUCONACETOBACTER-DIAZOTROPHICUS-LEVANSUCRASE; HETEROLOGOUS PROTEINS; EXPRESSION; YEAST; EXOPOLYSACCHARIDES; CLONING; SACB;

D O I：

10.1016/j.ijbiomac.2022.06.092

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

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.

引用

页码：203 / 211

页数：9

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