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题名: Molecular Cloning and Characterization of a Malic Enzyme Gene from the Oleaginous Yeast Lipomyces starkeyi
作者: Tang, Wei1, 2;  Zhang, Sufang1;  Tan, Haidong1;  Zhao, Zongbao K.1
关键词: Lipomyces starkeyi ;  Malic enzyme ;  RACE ;  Protein expression ;  Enzyme kinetics
刊名: MOLECULAR BIOTECHNOLOGY
发表日期: 2010-06-01
DOI: 10.1007/s12033-010-9255-8
卷: 45, 期:2, 页:121-128
收录类别: SCI
文章类型: Article
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
英文摘要: The malic enzyme-encoding cDNA (GQ372891) from the oleaginous yeast Lipomyces starkeyi AS 2.1560 was isolated, which has an 1719-bp open reading frame flanked by a 290-bp 5' untranslated sequence and a 92-bp 3' untranslated sequence. The proposed gene, LsME1, encoded a protein with 572 amino acid residues. The protein presented 58% sequence identity with the malic enzymes from Yarrowia lipolytica CLIB122 and Aspergillus fumigatus Af293. The LsME1 gene was cloned into the vector pMAL-p4x to express a fusion protein (MBP-LsME1) in Escherichia coli TB1. The fusion protein was purified and then cleaved by Factor Xa to give the recombinant LsME1. This purified enzyme took either NAD(+) or NADP(+) as the coenzyme but preferred NAD(+). The K (m) values for malic acid, NAD(+) and NADP(+) were 0.85 +/- A 0.05 mM, 0.34 +/- A 0.08 mM, and 7.4 +/- A 0.32 mM, respectively, at pH 7.3.
关键词[WOS]: LIPID-ACCUMULATION ;  ISOCITRATE DEHYDROGENASE ;  SACCHAROMYCES-CEREVISIAE ;  PIGEON LIVER ;  PURIFICATION ;  METABOLISM ;  ACID ;  EXPRESSION ;  OVEREXPRESSION ;  PROTEINS
语种: 英语
WOS记录号: WOS:000277343900004
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/141999
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.Chinese Acad Sci, Dalian Inst Chem Phys, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China

Recommended Citation:
Tang, Wei,Zhang, Sufang,Tan, Haidong,et al. Molecular Cloning and Characterization of a Malic Enzyme Gene from the Oleaginous Yeast Lipomyces starkeyi[J]. MOLECULAR BIOTECHNOLOGY,2010,45(2):121-128.
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