Improving the specific activity of beta-mannanase from Aspergillus niger BK01 by structure-based rational design

CORC > 天津工业生物技术研究所 > 结构生物学与蛋白酶学实验室郭瑞庭 > 期刊论文

	Improving the specific activity of beta-mannanase from Aspergillus niger BK01 by structure-based rational design
	Huang, Jian-Wen 1; Chen, Chun-Chi 2; Huang, Chun-Hsiang 2; Huang, Ting-Yung 3,4; Wu, Tzu-Hui 1; Cheng, Ya-Shan 3,4; Ko, Tzu-Ping 5; Lin, Cheng-Yen 3,4; Liu, Je-Ruei 1,6; Guo, Rey-Ting 2
刊名	BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS
	2014-03-01
卷号	1844 期号:3 页码:663-669
关键词	Aspergillus niger beta-Mannanase Crystal structure Rational design Thermophilic
英文摘要	beta-Mannanase has found various biotechnological applications because it is capable of degrading mannans into smaller sugar components. A highly potent example is the thermophilic beta-mannanase from Aspergillus niger BK01 (ManBK), which can be efficiently expressed in industrial yeast strains and is thus an attractive candidate for commercial utilizations. In order to understand the molecular mechanism, which helps in strategies to improve the enzyme's performance that would meet industrial demands, 3D-structural information is a great asset. Here, we present the 1.57 angstrom crystal structure of ManBK. The protein adopts a typical (beta/alpha)(8) fold that resembles the other GH5 family members. Polysaccharides were subsequently modeled into the substrate binding groove to identify the residues and structural features that may be involved in the catalytic reaction. Based on the structure, rational design was conducted to engineer ManBK in an attempt to enhance its enzymatic activity. Among the 23 mutants that we constructed, the most promising Y216W showed an 18 +/- 2.7% increase in specific activity by comparison with the wild type enzyme. The optimal temperature and heat tolerance profiles of Y216W were similar to those of the wild type, manifesting a preserved thermostability. Kinetic studies showed that Y216W has higher k(cat) values than the wild type enzyme, suggesting a faster turnover rate of catalysis. In this study we applied rational design to ManBK by using its crystal structure as a basis and identified the Y216W mutant that shows great potentials in industrial applications. (c) 2014 Elsevier B.V. All rights reserved.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Biochemistry & Molecular Biology ; Biophysics
研究领域[WOS]	Biochemistry & Molecular Biology ; Biophysics
关键词[WOS]	HYDROLASE FAMILY 5 ; CRYSTAL-STRUCTURE ; PICHIA-PASTORIS ; 3-DIMENSIONAL STRUCTURE ; FUNCTIONAL EXPRESSION ; SUBSTRATE-SPECIFICITY ; ENDO-BETA-1,4-MANNANASE ; CLONING ; COMPLEX ; CELLOTETRAOSE
收录类别	SCI
语种	英语
WOS记录号	WOS:000333491100021
内容类型	期刊论文
源URL	[http://124.16.173.210/handle/834782/1351]
专题	天津工业生物技术研究所_结构生物学与蛋白酶学实验室郭瑞庭_期刊论文
作者单位	1.Natl Taiwan Univ, Inst Biotechnol, Dept Anim Sci & Technol, Taipei 106, Taiwan 2.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Ind Enzymes Natl Engn Lab, Tianjin 300308, Peoples R China 3.Genozyme biotechnol Inc, Taipei 106, Taiwan 4.AsiaPac Biotechnol Co Ltd, Dongguan 523808, Peoples R China 5.Acad Sinica, Inst Biol Chem, Taipei 115, Taiwan 6.Acad Sinica, Agr Biotechnol Res Ctr, Taipei 115, Taiwan
推荐引用方式 GB/T 7714	Huang, Jian-Wen,Chen, Chun-Chi,Huang, Chun-Hsiang,et al. Improving the specific activity of beta-mannanase from Aspergillus niger BK01 by structure-based rational design[J]. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS,2014,1844(3):663-669.
APA	Huang, Jian-Wen.,Chen, Chun-Chi.,Huang, Chun-Hsiang.,Huang, Ting-Yung.,Wu, Tzu-Hui.,...&Guo, Rey-Ting.(2014).Improving the specific activity of beta-mannanase from Aspergillus niger BK01 by structure-based rational design.BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS,1844(3),663-669.
MLA	Huang, Jian-Wen,et al."Improving the specific activity of beta-mannanase from Aspergillus niger BK01 by structure-based rational design".BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS 1844.3(2014):663-669.