Identification of a novel fumarase C from Streptomyces lividans TK54 as a good candidate for malate production
文献类型：期刊论文
| 作者 | Su RR1; Wang A1; Hou ST1; Gao P1; Zhu GP[*]1; Wang W[*]2 |
| 刊名 | MOLECULAR BIOLOGY REPORTS
 |
| 出版日期 | 2014 |
| 卷号 | 41期号:1页码:497-504 |
| 通讯作者 | gpz1996@yahoo.com ; wwang@mail.kiz.ac.cn |
| 合作状况 | 其它 |
| 英文摘要 | Fumarase is a key enzyme that catalyzes the reversible hydration of fumarate to l-malate in the tricarboxylic acid cycle. This reaction has been extensively utilized for industrial applications in producing l-malate. In this study, a fumarase C gene from Streptomyces lividans TK54 (slFumC) was cloned and expressed as a fused protein (SlFumC) in Escherichia coli. The molecular mass of SlFumC was about 49 kDa determined by SDS-PAGE. Kinetic studies showed that the K (m) value of SlFumC for l-malate increased by approximately 8.5-fold at pH 6.5 (6.7 +/- A 0.81 mM) to 8.0 (57.0 +/- A 1.12 mM), which was higher than some known fumarases. The catalytic efficiency (k (cat)) and the specific activity increased by about 9.5-fold at pH 6.5 (65 s(-1)) to 8.0 (620 s(-1)) and from 79 U/mg at pH 6.5 to 752 U/mg at pH 8.0, respectively. Therefore, SlFumC may acquire strong catalytic ability by increasing pH to partially compensate for the loss of substrate affinity. The enzyme also showed substrate inhibition phenomenon, which is pH-dependent. Specific activity of SlFumC was gradually enhanced with increasing phosphate concentrations. However, no inhibition was observed at high concentration of phosphate ion, which was distinctly different in case of other Class II fumarases. In industrial process, the reaction temperatures for l-malate production are usually set between 40 and 60 A degrees C. The recombinant SlFumC displayed maximal activity at 45 A degrees C and remained over 85 % of original activity after 48 h incubation at 40 A degrees C, which was more thermostable than other fumarases from Streptomyces and make it an efficient enzyme for use in the industrial production of l-malate. |
| 收录类别 | SCI |
| 资助信息 | This research was supported by funds from the National High Technology Research and Development Program (‘‘863’’ Program: 2012AA02A708), the National Natural Science Foundation of China (31170005), Specialized Research Fund for the Doctoral Program of Higher Education of China (20113424110004), the Fund of State Key Laboratory of Genetics Resources and Evo- lution from Kunming Institute of Zoology (Chinese Academy of Sciences, CAS) (GRE-KF11-07) and Program for Innovative Research Team in Anhui Normal University. |
| 语种 | 英语 |
| WOS记录号 | WOS:000329100500054 |
| 公开日期 | 2014-01-24 |
| 源URL | [http://159.226.149.42:8088/handle/152453/7785]  |
| 专题 | 昆明动物研究所_基因起源组 昆明动物研究所_遗传资源与进化国家重点实验室 |
| 作者单位 | 1.Institute of Molecular Biology and Biotechnology, Anhui Normal University, No. 1 Beijing East Road, Wuhu 241000, Anhui, People’s Republic of China 2.State Key Laboratory of Genetics Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, No. 32 Jiaochang Donglu, Kunming 650223, Yunnan, China |
| 推荐引用方式 GB/T 7714 | Su RR,Wang A,Hou ST,et al. Identification of a novel fumarase C from Streptomyces lividans TK54 as a good candidate for malate production[J]. MOLECULAR BIOLOGY REPORTS,2014,41(1):497-504. |
| APA | Su RR,Wang A,Hou ST,Gao P,Zhu GP[*],&Wang W[*].(2014).Identification of a novel fumarase C from Streptomyces lividans TK54 as a good candidate for malate production.MOLECULAR BIOLOGY REPORTS,41(1),497-504. |
| MLA | Su RR,et al."Identification of a novel fumarase C from Streptomyces lividans TK54 as a good candidate for malate production".MOLECULAR BIOLOGY REPORTS 41.1(2014):497-504. |
入库方式： OAI收割
来源：昆明动物研究所
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