Expression and characterization of two secreted His(6)-tagged endo-beta-1,4-glucanases from the mollusc Ampullaria crossean in Pichia pastoris

	Expression and characterization of two secreted His(6)-tagged endo-beta-1,4-glucanases from the mollusc Ampullaria crossean in Pichia pastoris
	Guo, R; Ding, M; Zhang, SL; Xu, GJ; Zhao, FK
刊名	ACTA BIOCHIMICA ET BIOPHYSICA SINICA
	2008
卷号	40 期号:5 页码:419-425
关键词	cellulase endoglucanase EG27I EG27II Ampullaria crossean Pichia pastoris
通讯作者	Zhao, FK (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, Key Lab Prote, Shanghai 20031, Peoples R China.,fkzhao@sibs.ac.cn
英文摘要	Two endo-beta-1,4-glucanase cDNAs, eg27I and eg27II, from the mollusc Ampullaria crossean were expressed in Pichia pastoris cells. The secreted His(6)-tagged proteins were purified in a single chromatography step. The purified recombinant EG27I and EG27II showed enzymatic activity on carboxylmethyl cellulose sodium salt at 15.31 U/mg and 12.40 U/mg, respectively. The optimum pH levels of the recombinant EG27I and EG27II were 5.5 and 5.5-6.0, respectively, and the optimum temperatures were 50 degrees C and 50 degrees C-55 degrees C, respectively. The pH stability study revealed that both EG27I and EG27II showed their highest stability at pH 8.0. Analysis of their thermostability indicated that both EG27I and EG27II were relatively stable up to 40 degrees C. Site-directed mutagenesis of Asp43 and Asp153 of both EG27I and EG27II showed that the two Asp residues are critical for the enzymatic activity.
学科主题	Biochemistry & Molecular Biology; Biophysics
类目[WOS]	Biochemistry & Molecular Biology ; Biophysics
关键词[WOS]	MULBERRY LONGICORN BEETLE ; ENZYMATIC-ACTIVITY ; APRIONA-GERMARI ; CELLULASE GENE ; CDNA CLONING ; ENDOGENOUS CELLULASE ; MOLECULAR-CLONING ; ENDOGLUCANASE-V ; PURIFICATION ; TERMITE
收录类别	SCI
语种	英语
WOS记录号	WOS:000255930600007
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/1378]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Guo, R,Ding, M,Zhang, SL,et al. Expression and characterization of two secreted His(6)-tagged endo-beta-1,4-glucanases from the mollusc Ampullaria crossean in Pichia pastoris[J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA,2008,40(5):419-425.
APA	Guo, R,Ding, M,Zhang, SL,Xu, GJ,&Zhao, FK.(2008).Expression and characterization of two secreted His(6)-tagged endo-beta-1,4-glucanases from the mollusc Ampullaria crossean in Pichia pastoris.ACTA BIOCHIMICA ET BIOPHYSICA SINICA,40(5),419-425.
MLA	Guo, R,et al."Expression and characterization of two secreted His(6)-tagged endo-beta-1,4-glucanases from the mollusc Ampullaria crossean in Pichia pastoris".ACTA BIOCHIMICA ET BIOPHYSICA SINICA 40.5(2008):419-425.