Structural and biochemical insights into the degradation mechanism of chitosan by chitosanase OU01
文献类型：期刊论文
| 作者 | Lyu, Qianqian4; Shi, Yanhong1 ; Wang, Song2; Yang, Yan4; Han, Baoqin4; Liu, Wanshun4; Jones, David N. M.3; Liu, Weizhi4
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| 刊名 | BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS
 |
| 出版日期 | 2015-09 |
| 卷号 | 1850期号:9页码:1953-1961 |
| 关键词 | Chitosan Glycoside hydrolysis Degradation Mutagenesis X-ray crystallography |
| ISSN号 | 0304-4165 |
| DOI | 10.1016/j.bbagen.2015.06.011 |
| 文献子类 | Article |
| 英文摘要 | Background: A detailed knowledge about the degradation mechanism of chitosanase hydrolysis is critical for the design of novel enzymes to produce well-defined chito-oligosaccharide products. Methods: Through the combination of structural and biochemical analysis, we present new findings that provide novel insights into the degradation mechanism of chitosanase OU01. Results: We have determined the crystal structure of Asp(43)/Ala mutant of OU01, and have trapped the hydrolyzed product of the reaction. This structure reveals the role of the general acid (Glu(25)) in catalysis. Two structural features about the mechanisms of the non-processive chitosanases are described for the first time. 1). Structural comparison reveals that the enzyme goes through an open-closed-open conformational transition upon substrate binding and product release; 2). polar residues constitute the substrate binding cleft. Additional site important for polymeric substrate recognition is identified and a three-step polymeric substrate recognition mechanism is proposed. Conclusions: Detailed substrate recognition mechanism is described for non-processive chitosanase for the first time. General significance: These findings provide new structural insights into the understanding of overall hydrolysis mechanism for non-processive chitosanase, and also will facilitate the design of new enzymes used for industrial purpose. (C) 2015 Elsevier B.V. All rights reserved. |
| WOS关键词 | STREPTOMYCES SP N174 ; SERRATIA-MARCESCENS ; CRYSTAL-STRUCTURE ; SUBSTRATE RECOGNITION ; CATALYTIC CENTER ; GH19 CHITINASE ; PROCESSIVITY ; CHITOOLIGOSACCHARIDES ; RESIDUES ; BINDING |
| 资助项目 | Natural Science Foundation of Shandong Province[ZR2013CM044] ; SRF for ROCS, SEM[1792] ; National key technology R&D program of the Ministry of Science and Technology[2013BAB01B02] ; Qingdao Science and Technology projects[2013-12-007-SW] ; Zhenjiang Science and Technology project[2013C33192] |
| WOS研究方向 | Biochemistry & Molecular Biology ; Biophysics |
| 语种 | 英语 |
| WOS记录号 | WOS:000359173900034 |
| 出版者 | ELSEVIER SCIENCE BV |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/276426]  |
| 专题 | 分析化学研究室 |
| 通讯作者 | Liu, Weizhi |
| 作者单位 | 1.Chinese Acad Sci, Shanghai Inst Mat Med, Shanghai 200031, Peoples R China; 2.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Qingdao, Peoples R China; 3.Univ Colorado, Sch Med, Dept Pharmacol, Aurora, CO 80045 USA 4.Ocean Univ China, Coll Marine Life Sci, Qingdao, Peoples R China; |
| 推荐引用方式 GB/T 7714 | Lyu, Qianqian,Shi, Yanhong,Wang, Song,et al. Structural and biochemical insights into the degradation mechanism of chitosan by chitosanase OU01[J]. BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS,2015,1850(9):1953-1961. |
| APA | Lyu, Qianqian.,Shi, Yanhong.,Wang, Song.,Yang, Yan.,Han, Baoqin.,...&Liu, Weizhi.(2015).Structural and biochemical insights into the degradation mechanism of chitosan by chitosanase OU01.BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS,1850(9),1953-1961. |
| MLA | Lyu, Qianqian,et al."Structural and biochemical insights into the degradation mechanism of chitosan by chitosanase OU01".BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS 1850.9(2015):1953-1961. |
入库方式： OAI收割
来源：上海药物研究所
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