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Heterologous expression and biochemical characterization of a GHF9 endoglucanase from the termite Reticulitermes speratus in Pichia pastoris
Zhang,Pengfei1; Yuan,Xianghua1; Du,Yuguang2; Li,Jian-Jun2
2018-06-01
Source PublicationBMC Biotechnology
ISSN1472-6750
Volume18Issue:1
Abstract

AbstractBackgroundCellulases are of great significance for full utilization of lignocellulosic biomass. Termites have an efficient ability to degrade cellulose. Heterologous production of the termite-origin cellulases is the first step to realize their industrial applications. The use of P. pastoris for the expression of recombinant proteins has become popular. The endoglucanase from Reticulitermes speratus (RsEG), belonging to glycoside hydrolase family 9 (GHF9), has not been produced in P. pastoris yet.ResultsA mutant RsEGm (G91A/Y97W/K429A) was successfully overexpressed in P. pastoris. RsEGm, with optimum pH?5.0, was active over the pH range of 4.0 to 9.0, and exhibited superior pH stability over between pH?4.0 and pH?11.0. It displayed the highest activity and good stability at 40?°C, but lost activity quickly at 50?°C. The apparent kinetic parameters of RsEGm against Carboxymethyl Cellulose (CMC) were determined, with Km and Vmax of 7.6?mg/ml and 5.4 μmol/min?mg respectively. Co2+, Mn2+ and Fe2+ enhanced the activity of RsEGm by 32.0, 19.5 and 11.2% respectively, while Pb2+ and Cu2+ decreased its activity by 19.6 and 12.7% separately.ConclusionsRsEGm could be overexpressed in P. pastoris. It was stable between pH?4.0 and pH?11.0, and exhibited higher stability at temperatures ≤ 40?°C. This endoglucanase may have potential to be used in the field of laundry, textile and lignocellulose-based biofuels and chemicals.

KeywordReticulitermes Speratus Ghf9 Endoglucanse Heterologous Expression Pichia Pastoris Enzymology
DOI10.1186/s12896-018-0432-3
Language英语
WOS IDBMC:10.1186/s12896-018-0432-3
PublisherBioMed Central
Citation statistics
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/24493
Collection中国科学院过程工程研究所
Corresponding AuthorYuan,Xianghua; Du,Yuguang; Li,Jian-Jun
Affiliation1.Sichuan Normal Univ, Coll Life Sci, Chengdu 610101, Sichuan, Peoples R China
2.Chinese Acad Sci, Natl Key Lab Biochem Engn, Natl Engn Res Ctr Biotechnol Beijing, Key Lab Biopharmaceut Prod & Formulat Engn,PLA,In, 1 North 2nd St, Beijing 100190, Peoples R China
Recommended Citation
GB/T 7714
Zhang,Pengfei,Yuan,Xianghua,Du,Yuguang,et al. Heterologous expression and biochemical characterization of a GHF9 endoglucanase from the termite Reticulitermes speratus in Pichia pastoris[J]. BMC Biotechnology,2018,18(1).
APA Zhang,Pengfei,Yuan,Xianghua,Du,Yuguang,&Li,Jian-Jun.(2018).Heterologous expression and biochemical characterization of a GHF9 endoglucanase from the termite Reticulitermes speratus in Pichia pastoris.BMC Biotechnology,18(1).
MLA Zhang,Pengfei,et al."Heterologous expression and biochemical characterization of a GHF9 endoglucanase from the termite Reticulitermes speratus in Pichia pastoris".BMC Biotechnology 18.1(2018).
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