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题名: Purification and characterization of a beta-1,3-glucomannanase expressed in Pichia pastoris
作者: Yang, Fan1, 3;  Zhang, Sufang1;  Jin, Guojie1, 3;  Lin, Xinping1, 3;  Zhao, Zongbao K.1, 2
关键词: beta-1,3-glucomannanase ;  Secretory expression ;  Pichia pastoris ;  Cell wall ;  Enzyme activity
刊名: ENZYME AND MICROBIAL TECHNOLOGY
发表日期: 2011-07-10
DOI: 10.1016/j.enzmictec.2011.04.005
卷: 49, 期:2, 页:223-228
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine
类目[WOS]: Biotechnology & Applied Microbiology
研究领域[WOS]: Biotechnology & Applied Microbiology
英文摘要: The glycoside hydrolase beta-1,3-glucomannanase is an enzyme that specifically breaks the beta-1,3 glycosidic bond of the glucomannan, the main cell wall constituent of some yeasts. In this work, a codon optimized DNA sequence of the MAN5C gene from Penicillium Iilacinum ATCC 36010 was expressed in the yeast Pichia pastoris under the control of AOX1 promoter. The recombinant protein pIMAN5C was purified from the shake flask culture and the stirred-tank bioreactor culture in yields of 30.0 mg/l and 224.0 mg/l, respectively. The purified protein had a specific activity of 14.6 U/mg at 37 degrees C, pH 4.5. Biochemical analysis showed that the optimal temperature and pH for pIMAN5C were 50 degrees C and 4.5, respectively. The recombinant pIMAN5C was efficient in lysis of the cell wall of the red yeast Rhodosporidium toruloides to form protoplast. Our work provided an effective system for heterogeneous production of beta-1,3-glucomannanase, which should facilitate a more convenient application of this enzyme in biotechnology and other related areas. (C) 2011 Elsevier Inc. All rights reserved.
关键词[WOS]: YEAST-CELL WALL ;  LYTIC ENZYME ;  RHODOSPORIDIUM-TORULOIDES ;  SACCHAROMYCES-CEREVISIAE ;  SCHIZOPHYLLUM-COMMUNE ;  PROTOPLAST FORMATION ;  GENE ;  IDENTIFICATION
语种: 英语
WOS记录号: WOS:000293439100017
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/142544
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.CAS, Dalian Inst Chem Phys, Div Biotechnol, Dalian 116023, Peoples R China
2.Dalian Natl Lab Clean Energy, Dalian 116023, Peoples R China
3.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China

Recommended Citation:
Yang, Fan,Zhang, Sufang,Jin, Guojie,et al. Purification and characterization of a beta-1,3-glucomannanase expressed in Pichia pastoris[J]. ENZYME AND MICROBIAL TECHNOLOGY,2011,49(2):223-228.
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