Enhanced expression of the codon-optimized exo-inulinase gene from the yeast Meyerozyma guilliermondii in Saccharomyces sp W0 and bioethanol production from inulin

被引:37
作者
Liu, Guang-Lei [1 ]
Fu, Ge-Yi [1 ]
Chi, Zhe [1 ]
Chi, Zhen-Ming [1 ]
机构
[1] Ocean Univ China, UNESCO Chinese Ctr Marine Biotechnol, Qingdao 266003, Peoples R China
基金
中国国家自然科学基金;
关键词
Exo-inulinase; Meyerozyma guilliermondii; Saccharomyces sp W0; Codonoptimization; Ethanol; Inulin; JERUSALEM-ARTICHOKE TUBERS; PICHIA-GUILLIERMONDII; ETHANOL-PRODUCTION; KLUYVEROMYCES-MARXIANUS; DIRECT CONVERSION; PROTEIN; USAGE; ENDOINULINASE; FERMENTATION; HYDROLYSIS;
D O I
10.1007/s00253-014-6079-7
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
In the present study, after the exo-inulinase gene INU1 from Meyerozyma guilliermondii was optimized according to the codon usage bias of Saccharomyces cerevisiae, both the optimized gene INU1Y and the native gene INU1 were ligated into the homologous integration expression vector pMIRSC11 and expressed in Saccharomyces sp. W0. It was determined that the inulinase activity of the recombinant yeast Y13 with the optimized gene INU1Y was 43.84 U/mL, which was obviously higher than that (31.39 U/mL) produced by the recombinant yeast EX3 with the native gene INU1. Moreover, it was indicated that the recombinant yeast Y13 could produce 126.30 mg/mL ethanol from 300.0 g/L inulin while the recombinant yeast EX3 and Saccharomyces sp. W0 produced 122.75 mg/mL and 114.15 mg/mL ethanol, respectively, under the same conditions. In addition, the ethanol productivity of the recombinant yeast Y13 was 2.25 mg/mL/h within 48 h of the fermentation, which was obviously higher than that of the recombinant yeast EX3 (1.97 mg/mL/h) and Saccharomyces sp. W0 (1.77 mg/mL/h) within the same period. The results demonstrated that the recombinant yeast Y13 had higher ethanol production and productivity than the recombinant yeast EX3 and Saccharomyces sp. W0. Therefore, it was concluded that the codon optimization of the exo-inulinase gene from M. guilliermondii effectively enhanced inulinase activity and improved ethanol production from inulin by Saccharomyces sp. W0 carrying the optimized inulinase gene.
引用
收藏
页码:9129 / 9138
页数:10
相关论文
共 35 条
[1]   Codon usage: Nature's roadmap to expression and folding of proteins [J].
Angov, Evelina .
BIOTECHNOLOGY JOURNAL, 2011, 6 (06) :650-659
[2]   Biotechnological potential of inulin for bioprocesses [J].
Chi, Zhen-Ming ;
Zhang, Tong ;
Cao, Tian-Shu ;
Liu, Xiao-Yan ;
Cui, Wei ;
Zhao, Chun-Hai .
BIORESOURCE TECHNOLOGY, 2011, 102 (06) :4295-4303
[3]  
CHI ZM, 1993, BIOTECHNOL LETT, V15, P877
[4]   Direct conversion of inulin into single cell protein by the engineered Yarrowia lipolytica carrying inulinase gene [J].
Cui, Wei ;
Wang, Qi ;
Zhang, Fang ;
Zhang, Shi-Cui ;
Chi, Zhen-Ming ;
Madzak, Catherine .
PROCESS BIOCHEMISTRY, 2011, 46 (07) :1442-1448
[5]  
Gietz R Daniel, 2006, Methods Mol Biol, V313, P107
[6]   Inulinase production by a marine yeast Pichia guilliermondii and inulin hydrolysis by the crude inulinase [J].
Gong, Fang ;
Sheng, Jun ;
Chi, Zhenming ;
Li, Jing .
JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY, 2007, 34 (03) :179-185
[7]   Purification and Characterization of Extracellular Inulinase from a Marine Yeast Pichia guilliermondii and Inulin Hydrolysis by the Purified Inulinase [J].
Gong, Fang ;
Zhang, Tong ;
Chi, Zhenming ;
Sheng, Jun ;
Li, Jing ;
Wang, Xianghong .
BIOTECHNOLOGY AND BIOPROCESS ENGINEERING, 2008, 13 (05) :533-539
[8]   Codon bias and heterologous protein expression [J].
Gustafsson, C ;
Govindarajan, S ;
Minshull, J .
TRENDS IN BIOTECHNOLOGY, 2004, 22 (07) :346-353
[9]   Enhanced expression of endoinulinase from Aspergillus niger by codon optimization in Pichia pastoris and its application in inulooligosaccharide production [J].
He, Miao ;
Wu, Dan ;
Wu, Jing ;
Chen, Jian .
JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY, 2014, 41 (01) :105-114
[10]   Thermotolerant Kluyveromyces marxianus and Saccharomyces cerevisiae strains representing potentials for bioethanol production from Jerusalem artichoke by consolidated bioprocessing [J].
Hu, Nan ;
Yuan, Bo ;
Sun, Juan ;
Wang, Shi-An ;
Li, Fu-Li .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2012, 95 (05) :1359-1368