Cloning of a novel levoglucosan kinase gene from Lipomyces starkeyi and its expression in Escherichia coli
文献类型：期刊论文
| 作者 | Dai, Jianghong; Yu, Zhisheng; He, Yongzhi; Zhang, Ling; Bai, Zhihui; Dong, Zhiyang; Du, Yuguo; Zhang, Hongxun |
| 刊名 | WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
 |
| 出版日期 | 2009-09 |
| 卷号 | 25期号:9页码:1589-1595 |
| 关键词 | Levoglucosan Kinase Gene Lipomyces Starkeyi Yz-215 Race Cellulose Pyrolysis |
| 英文摘要 | Levoglucosan, cellulosic pyrolysate, is converted to glucose-6-phosphate by a specific levoglucosan kinase in fungi. A novel cDNA of levoglucosan kinase gene (lgk) from yeast Lipomyces starkeyi YZ-215 was isolated by RACE method. The 1,445 bp cDNA fragment (lgk) harbouring the kinase gene exhibited one open reading frame (ORF) composed of 1,317 bp flanked by a 14 bp 5'-UTR and a 114 bp 3'-UTR, including a 25 bp poly(A) tail. The ORF encoded a 439 amino acid polypeptide with a 48.4 kDa predicted molecular mass. Analysis of amino sequence revealed that the kinase belonged to the bacterial anhydro-N-acetylmuramic acid kinase (AnmK) family, and kinase-like proteins existed in some fungi, especially in filamentous fungi such as Aspergillus. The kinase gene was transformed into Escherichia coli BL21 (DE3), recombinant E. coli could grow in M9 minimal medium with levoglucosan as a sole carbon source when induced by IPTG. In addition, the recombinant kinase was overexpressed, purified and characterized. The kinase was stable at pH 7-10 and showed maximum activity at 30A degrees C and pH 9.0 as natural kinase, but presented higher thermostability. Kinetic constants (apparent K (m) values) for LG and ATP were 105.3 +/- A 12.5 and 0.20 +/- A 0.02 mM, respectively. Furthermore, the kinase showed substrate specificity for LG. This novel levoglucosan kinase gene would be useful in constructing recombinant microbial strains for the efficient bioconversion of cellulosic pyrolysate to ethanol. |
| WOS记录号 | WOS:000268729600010 |
| 源URL | [http://ir.rcees.ac.cn/handle/311016/21807]  |
| 专题 | 生态环境研究中心_中国科学院环境生物技术重点实验室 |
| 推荐引用方式 GB/T 7714 | Dai, Jianghong,Yu, Zhisheng,He, Yongzhi,et al. Cloning of a novel levoglucosan kinase gene from Lipomyces starkeyi and its expression in Escherichia coli[J]. WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY,2009,25(9):1589-1595. |
| APA | Dai, Jianghong.,Yu, Zhisheng.,He, Yongzhi.,Zhang, Ling.,Bai, Zhihui.,...&Zhang, Hongxun.(2009).Cloning of a novel levoglucosan kinase gene from Lipomyces starkeyi and its expression in Escherichia coli.WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY,25(9),1589-1595. |
| MLA | Dai, Jianghong,et al."Cloning of a novel levoglucosan kinase gene from Lipomyces starkeyi and its expression in Escherichia coli".WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY 25.9(2009):1589-1595. |
入库方式： OAI收割
来源：生态环境研究中心
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