生物学杂志 ›› 2022, Vol. 39 ›› Issue (5): 1-.doi: 10.3969/j.issn.2095-1736.2022.05.001

• 研究报告 •    下一篇

吩嗪合成蛋白PhzA和PhzB的功能比较

  

  1. 上海交通大学 生命科学技术学院 微生物代谢国家重点实验室, 上海 200240
  • 出版日期:2022-10-18 发布日期:2022-10-18
  • 通讯作者: 张雪洪,教授,博士生导师,研究方向为微生物学和生物工程,E-mail: xuehzhang@sjtu.edu.cn
  • 作者简介:孔德毓,硕士研究生,研究方向为代谢工程,E-mail: 940107973@qq.com
  • 基金资助:
    国家自然科学基金项目(No.32070051)

Functional comparison of phenazine synthetic proteins PhzA and PhzB

  1. State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology,
    Shanghai Jiao Tong University, Shanghai 200240, China
  • Online:2022-10-18 Published:2022-10-18

摘要: 为研究PhzA/B蛋白之间的功能差异及其对吩嗪核心前体(PDC和PCA)合成的影响,通过比对、筛选并合成不同来源的phzA/B基因,发现在异源phzB基因替换内源phzAB基因突变株的代谢产物中都检测到PDC和PCA的合成;而通过异源phzA基因替换内源phzA基因及异源phzB基因替换内源phzB基因的突变株,其代谢产物中只检测到PCA的存在,且phzAB的同时存在会促进PCA的大量合成。对PhzA/B蛋白的结构分析发现:不同来源的PhzB蛋白结构中部分区域存在一定差异,PhzA与PhzB的蛋白结构之间相似性较高,但PhzA缺少相应的PhzB活性氨基酸位点。不同来源的PhzA/B蛋白序列相似性较高,在结构与功能上有一定差异,都在吩嗪类化合物的合成过程中起着重要的作用,其中PhzB蛋白促进PDC的催化合成,而PhzA蛋白促进PCA的合成。

关键词: 吩嗪合成蛋白, 吩嗪核心前体, 代谢工程, 系统分析, 生物合成

Abstract: In order to explore the functional differences between PhzA/B and the effects on the synthesis of PDC and PCA, after comparing, screening and synthesizing the phzA and phzB genes from different bacteria were studied. The result showed that PDC and PCA could be detected in seven mutants, in which heterologous phzB genes replaced the endogenous phzA and phzB gene at the same time. However, PDC could not be detected in the secondary metabolites of five mutants, in which two heterologous phzA genes replaced the endogenous phzA gene and three heterologous phzB genes replaced the endogenous phzB gene, and the simultaneous presence of phzAB wound promote the synthesis of PCA. Moreover, structural analysis showed that there were some differences in some regions of PhzB proteins from different bacteria. The protein structures of PhzA and PhzB were similar, but PhzA lacked the corresponding active amino acid sites of PhzB. In conclusion, PhzA/B proteins from different bacteria had relatively high sequence similarity, but had certain differences in structure and function. They all played an important role in the synthesis of phenazine compounds. Among them, the function of PhzB protein was important to catalyze the synthesis of PDC, while PhzA protein could promote the synthesis of PCA.

Key words: phenazine synthetic protein, phenazine core precursor, metabolic engineering, systems analysis, biosynthesis

中图分类号: