绿针假单胞菌HT66中luxR_19对吩嗪-1-甲酰胺合成的影响

doi:10.13560/j.cnki.biotech.bull.1985.2015.08.024

生物技术通报 ›› 2015, Vol. 31 ›› Issue (8): 166-173.doi: 10.13560/j.cnki.biotech.bull.1985.2015.08.024

绿针假单胞菌HT66中luxR_19对吩嗪-1-甲酰胺合成的影响

陈亚汶, 彭华松, 王威, 张雪洪

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

收稿日期:2014-12-08 出版日期:2015-08-21 发布日期:2015-08-22
作者简介:陈亚汶,女,硕士研究生,研究方向：微生物代谢调控;E-mail：moda_yaya@hotmail.com
基金资助:
国家自然科学基金项目（31270084）,国家“863”计划项目（2012AA022107）

The Effects of luxR_19 on the Synthesis of Phenzaine-1-carboxamide in Pseudomonas chlororaphis HT66

Chen Yawen, Peng Huasong, Wang Wei, Zhang Xuehong

State Key Laboratory of Microbial Metabolism,School of Life Sciences and Biotechnology,Shanghai Jiao Tong University,Shanghai 200240

Received:2014-12-08 Published:2015-08-21 Online:2015-08-22

摘要/Abstract

摘要： 旨在研究调控基因luxR_19对绿针假单胞菌HT66中的抑菌物质吩嗪-1-甲酰胺（phenazine-1-carboxamide,PCN）合成及菌体生长的影响,通过分子操作构建了luxR_19基因敲除株、回补菌株、过表达菌株以及luxR_19（G85A）点突变菌株,检测了目标菌株生长曲线、PCN发酵结果、细菌泳动性与从动性等。结果显示,将luxR_19敲除后PCN产量明显降低,对luxR_19基因进行过表达,发现能够将PCN产量提高2倍;luxR_19对于吩嗪合成的多个调控基因以及合成基因表达均产生影响;luxR_19的敲除对HT66的生长及泳动性不产生影响,而对细菌的从动性有一定的促进作用;luxR_19（G85A）点突变菌株吩嗪产量较野生型相差很小。结果表明,luxR_19对PCN的合成为正调控基因,一定程度上影响HT66的从动性,第254位碱基突变对PCN合成产量不产生影响。

关键词: 绿针假单胞菌, HT66, 吩嗪-1-甲酰胺, LuxR家族转录调控因子

Abstract: In order to study the potential effects of regulator gene luxR_19 on the synthesis of antibacterial substance phenazine-1-carboxamide（PCN）and physiological-biochemical characteristics in Pseudomonas chlororaphis HT66, strains of gene luxR_19-knockout, -complementation, -overexpression and -point-mutation were constructed, and then the growth curve, fermentation production of PCN, swimming and swarming activities were tested at the target strains. The results showed that the knockout of luxR_19 lowered the production of PCN and the over-expression of gene luxR_19 increased the double production of PCN compared to the wild type;luxR_19 gene affected the expression of several genes relating to the regulation or synthesis of phenazines. Besides, the knockout of luxR_19 had no effects on the growth and swimming activity, however, promoted the swarming activity of HT66. The production of PCN in point-mutation strain of luxR_19（G85A）was similar to that in wild type. The above results indicated that luxR_19 was the positive regulator of PCN production, and could affect the swarming activity of HT66. Besides, the point-mutation at 254^th site in luxR_19 did not affect the production of PCN.

Key words: Pseudomonas chlororaphis, HT66, phenazine-1-carboxamide, transcriptional regulator of LuxR family

陈亚汶, 彭华松, 王威, 张雪洪. 绿针假单胞菌HT66中luxR_19对吩嗪-1-甲酰胺合成的影响[J]. 生物技术通报, 2015, 31(8): 166-173.

Chen Yawen, Peng Huasong, Wang Wei, Zhang Xuehong. The Effects of luxR_19 on the Synthesis of Phenzaine-1-carboxamide in Pseudomonas chlororaphis HT66[J]. Biotechnology Bulletin, 2015, 31(8): 166-173.

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绿针假单胞菌HT66中luxR_19对吩嗪-1-甲酰胺合成的影响

The Effects of luxR_19 on the Synthesis of Phenzaine-1-carboxamide in Pseudomonas chlororaphis HT66

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