米曲霉6-磷酸葡萄糖酸脱氢酶基因(<em>gnd</em>)的克隆及生物信息学分析

吴晶晶; 洪志宏; 张卡; 陈宏文

米曲霉6-磷酸葡萄糖酸脱氢酶基因(gnd)的克隆及生物信息学分析

1.
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:

教育部留学回国人员科研启动基金(2010);福建省高等学校新世纪优秀人才支持计划(07FJRCO3);中央高校基本科研业务费专项资金(JB-ZR1112)

详细信息

通讯作者:
陈宏文(1969-),女,副教授,研究方向:生物工程、基因工程(E-mail:chenhw@hqu.edu.cn)

中图分类号: S 852
计量
- 文章访问数: 2849
- HTML全文浏览量: 116
- PDF下载量: 391
- 被引次数: 0
出版历程
- 收稿日期: 2012-04-05
- 修回日期: 2012-04-16
- 刊出日期: 2012-04-30

Cloning and Bioinformatic Analysis of 6-Phogluconate Dehydrogenase Gene from Aspergillus oryzae

1.
Department of Bioengineering & Biotechnology, College of Chemical Engineering, Hua Qiao University, Xiamen ,Fujian 361021, China

摘要

摘要: 采用PCR技术从米曲霉CICC2012菌株基因组中克隆6-磷酸葡萄糖酸脱氢酶基因(gnd),并利用生物信息学手段对其氨基酸序列、进化树、理化性质、蛋白质结构等进行分析。序列测定和分析结果表明,gnd基因序列长为1 723 bp,包含1个1 551 bp的开放阅读框,编码516个氨基酸;gnd基因编码的6PGDH氨基酸序列与黄曲霉6PGDH基因的同源性为99%,存在的丝氨酸、苏氨酸和酪氨酸磷酸化位点分别有11、2和6个;6PGDH蛋白分子量为57.3 kD,等电点为5.63;gnd基因编码蛋白二级结构α-螺旋区域占44.57%,β-折叠区域占12.79%,无规则卷曲区域占42.64%;氨基酸残基11~195位点为NADP+结合区域。
- 米曲霉 /
- 6-磷酸葡萄糖酸脱氢酶 /
- 克隆 /
- 生物信息学
Abstract: 6-Phogluconate dehydrogenase (6PGDH, EC 1.1.1.44)is one of the key enzymes in the pentose phosphate pathway(PPP). In this study, the 6-phogluconate dehydrogenase gene (gnd) of Aspergillus oryzae CICC2012 was cloned by means of PCR. Subsequently, the bioinformatic methodology was applied to determine the amino acid sequence homology, phylogenetic trees, physical-chemical properties and protein structure of the gene. The results revealed the gene’s length to be 1 723 bp, which encompassed an open reading frame with 1 551 bp encoding 516 amino acids. The 6PGDH encoded by gnd showed a 99% homology with the 6PGDH gene of Aspergillus flavus, which had 11 serine phosphorylation sites, 2 threonine phosphorylation sites and 6 tyrosine phosphorylation sites. The 6PGDH had a molecular weight of 57.3 kD and an isoelectric point of 5.63. The secondary structure of the gnd protein was 44.57% alpha helix, 12.79% beta sheet and 42.64% random coil. The 11-195 amino acid residues appeared to be the NADP+ binding sites.
- Aspergillus oryzae /
- 6-phosphate glucose dehydrogenase /
- cloning /
- bioinformatics

HTML全文

参考文献(4)

[1]	黄骥,王建飞,张红生.植物戊糖磷酸途径及两个关键酶的研究进展[J].植物学通报, 2004, 21(2): 139-145.
[2]	CHRISTOPHE D,ELIANA R,STEFANIA H,et al.Synthesis and biological evaluation of substrate-based inhibitors of 6-phosphogluconate dehydrogenase as potential drugs against African Trypanosomiasis[J]. Bioorganic & Medicinal Chemistry, 2003, 11: 3205-3214.
[3]	JUNKO O, RITSUKO K, SATOSHI M, et al. A novel gnd mutation leading to increased L L-lysine production in Corynebacterium glutamicum[J]. FEMS Microbiology Letters, 2005, 242: 265-274.
[4]	袁洪水,李术娜,张爱莲,等.石英砂研磨法快速提取顶头孢霉染色体DNA[J].河北农业大学学报, 2007, 30(5): 8-11.