Heterologous Expression and Characterization of a GH3 beta-Glucosidase from Thermophilic Fungi Myceliophthora thermophila in Pichia pastoris

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	Heterologous Expression and Characterization of a GH3 beta-Glucosidase from Thermophilic Fungi Myceliophthora thermophila in Pichia pastoris
	Zhao, Junqi; Guo, Chao; Tian, Chaoguang; Ma, Yanhe
刊名	APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
	2015-09-01
卷号	177 期号:2 页码:511-527
关键词	Myceliophthora thermophila beta-Glucosidase Substrate specificity Transglycosylation activity Pichia pastoris
英文摘要	A novel beta-glucosidase of glycoside hydrolase (GH) family 3 from Myceliophthora thermophila (mtbgl3b) was successfully expressed in Pichia pastoris. The full-length gene consists of 2613 bp nucleotides encoding a protein of 870 amino acids. MtBgl3b showed maximum activity at pH 5.0 and remained more than 70 % relative activity at 3.5-6.0. The enzyme displayed the highest activity at 60 A degrees C and kept about 90 % relative activity for 50-65 A degrees C; besides, the enzyme showed psychrophilic trait and remains 51 % relative activity at 40 A degrees C. MtBgl3b exhibited good stability over a wide pH range of 3.0-10.0 and was thermostable at 60 and 65 A degrees C. The enzyme displayed highest activity towards p-nitrophenyl-beta-d-glucopyranoside (pNPG), followed by p-nitrophenyl-d-cellobioside (pNPC), cellotetraose, cellotriose, cellobiose, and gentiobiose. When using 10 % cellobiose (w/v) as the substrate, the enzyme showed transglycosylation activity to produce the cellotriose. The kinetic parametric of K (m) and V (max) were 2.78 mM and 927.9 mu M mg(-1) min(-1), respectively. Finally, the reaction mode of the enzyme and the substrates were analyzed by molecular docking approach.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
研究领域[WOS]	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
关键词[WOS]	FAMILY 3 ; PAECILOMYCES-THERMOPHILA ; FUNCTIONAL EXPRESSION ; THERMOSTABLE ENZYMES ; TRICHODERMA-REESEI ; ASPERGILLUS-NIGER ; CLONING ; GENE ; PURIFICATION ; HYDROLYSIS
收录类别	SCI
语种	英语
WOS记录号	WOS:000360076200016
内容类型	期刊论文
源URL	[http://124.16.173.210/handle/834782/1519]
专题	天津工业生物技术研究所_微生物功能基因组学与生物技术实验室田朝光_期刊论文
作者单位	Chinese Acad Sci, Tianjin Airport Econ Area, Tianjin Inst Ind Biotechnol, Key Lab Syst Microbial Biotechnol, Tianjin 300308, Peoples R China
推荐引用方式 GB/T 7714	Zhao, Junqi,Guo, Chao,Tian, Chaoguang,et al. Heterologous Expression and Characterization of a GH3 beta-Glucosidase from Thermophilic Fungi Myceliophthora thermophila in Pichia pastoris[J]. APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY,2015,177(2):511-527.
APA	Zhao, Junqi,Guo, Chao,Tian, Chaoguang,&Ma, Yanhe.(2015).Heterologous Expression and Characterization of a GH3 beta-Glucosidase from Thermophilic Fungi Myceliophthora thermophila in Pichia pastoris.APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY,177(2),511-527.
MLA	Zhao, Junqi,et al."Heterologous Expression and Characterization of a GH3 beta-Glucosidase from Thermophilic Fungi Myceliophthora thermophila in Pichia pastoris".APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY 177.2(2015):511-527.