米曲霉磷酸果糖激酶基因克隆及序列分析

张卡; 杜钰; 陈宏文

doi:10.19303/j.issn.1008-0384.2013.06.016

米曲霉磷酸果糖激酶基因克隆及序列分析

doi: 10.19303/j.issn.1008-0384.2013.06.016

华侨大学化工学院生物工程与技术系,福建厦门,361021

基金项目:

中央高校基本科研业务费专项资金项目(JB-ZR1112)

福建省自然科学基金(2013)

华侨大学科研基金项目(12BS132)

详细信息

作者简介:
张卡(1987-),男,硕士研究生,研究方向:代谢工程(E-mail:zhang-karl@163.com);陈宏文(1969-),女,副教授,硕士生导师,研究方向:基因工程及代谢工程(E-mail:chenhw@hqu.edu.cn)

中图分类号: Q785
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- 被引次数: 0
出版历程
- 收稿日期: 2013-04-17
- 刊出日期: 2013-06-18

Cloning and Sequence Analysis Phosphofructokinase Gene from Aspergillus oryzae

Department of Bioengineering and Biotechnology,College of Chemical Engineering,Huaqiao University,Xiamen,Fujian 361021,China

摘要

摘要: 磷酸果糖激酶(PFK)是糖酵解途径的重要调控酶。以酿酒米曲霉CICC2012为材料,克隆获得PFK基因pfkA(GenBank登录号:KC113503.1)。序列分析表明:pfkA序列长度为2 722bp,含有2个内含子,开放阅读框长2 358bp;PFK为785个氨基酸组成的亲水蛋白,分子量86.0kDa,等电点6.38,含有2个PFK家族指纹结构;二级结构包含42.68%的α-螺旋,14.27%的β-折叠和43.06%的无规则卷曲;同源建模三级结构PFK含有N端和C端的2个结构域,底物果糖-6-磷酸结合于N端结构域。采用进化树分析,发现米曲霉PFK与丝状真菌PFK亲缘性较近。
- 磷酸果糖激酶 /
- 米曲霉 /
- 克隆 /
- 序列分析
Abstract: Phosphofructokinase(PFK)is one of the important regulatory enzymes in the Embden Meyerhof Parnas pathway.The PFK gene(pfkA)was cloned from the wine-brewing Aspergillus oryzae,CICC2012.It was subsequently sequenced and submitted to Genbank(accession number:KC113503.1)for verification.The sequence analysis revealed that pfkA was 2,722bp in length containing two introns,and 2,358bp long with an open reading frame encoding 785amino acids.The PFK was a hydrophilic protein containing two family signatures with a molecular weight of 86.0kD and an isoelectric point of 6.38.Its secondary structure consisted of 42.68% α-helix,14.27%β-folding and 43.06% random coil.The 3Dhomology model showed two(i.e.,N-and C-terminal) domains with the substrate,fructose-6-phosphate,bound to the N-terminal domain.The phylogenetic tree indicated pfkA′s close relationship with PFKs from other filamentous fungi.
- phosphofructokinase /
- Aspergillus oryzae /
- cloning /
- sequence analysis

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