食品科学 ›› 2017, Vol. 38 ›› Issue (10): 130-136.doi: 10.7506/spkx1002-6630-201710022

• 生物工程 • 上一篇    下一篇

1 株产烟酸羟基化酶海洋细菌H9的分离鉴定及 培养条件优化

株产烟酸羟基化酶海洋细菌H9的分离鉴定及培养条件优化陈国忠,郭秋翠,谭其秀,武硕军,刘炼强,冯志彬   

  1. 鲁东大学生命科学学院,山东 烟台 264025
  • 出版日期:2017-05-25 发布日期:2017-05-23
  • 基金资助:
    山东省现代农业产业技术体系家禽产业创新项目(SDAIT-11-10);城新创新基金项目(CXJ-06); 鲁东大学科研基金项目(27710301)

Isolation, Identification and Culture Optimization of Pseudomonas putida H9, a Marine Bacterium Producing Nicotinic Acid Hydroxylase

CHEN Guozhong, GUO Qiucui, TAN Qixiu, WU Shuojun, LIU Lianqiang, FENG Zhibin   

  1. School of Life Science, Ludong University, Yantai 264025, China
  • Online:2017-05-25 Published:2017-05-23

摘要: 利用96 孔板和紫外显影相结合的定向快速筛选方法,从烟台渤海湾采集的腐败鱼尸上分离筛选得到1 株高 产烟酸羟基化酶的菌株H9,通过形态、生理生化特征和16S rDNA系统发育分析鉴定为恶臭假单胞菌(Pseudomonas putida),对其产酶培养条件进行优化,最适培养基和培养条件为:可溶性淀粉15 g/L、玉米浆15 g/L、烟酸10 g/L、 K2HPO4 1.0 g/L、初始pH 7.0、培养温度25 ℃、接种量7%、500 mL三角瓶装液量50 mL。优化后烟酸羟基化酶活 力达到0.37 U/mL,比优化前提高了68%,采用静息细胞转化生产6-羟基烟酸产率达到124.77 g/L,具有工业化生产 6-羟基烟酸的应用前景。

关键词: 烟酸, 6-羟基烟酸, 恶臭假单胞菌, 烟酸羟基化酶, 培养条件优化

Abstract: A nicotinic acid hydroxylase- producing strain from spoiled fish from Bohai Bay in Yantai, designated H9, was rapidly and directionally screened using a 96-well microplate and ultraviolet imaging. Strain H9 was identified as Pseudomonas putida based on its morphological, physiological and biochemical characteristics, and 16S rDNA sequence. Our results showed that the highest activity of nicotinic acid hydroxylase of 0.37 U/mL was obtained by culturing strain H9 with 50 mL of a medium consisting of 15 g/L soluble starch, 15 g/L corn steep liquor, 10 g/L nicotinic acid and, 1.0 g/L K2HPO4 at an initial pH of 7.0 in a 500-mL flask at 25 ℃ with an inoculum amount of 7%, which was increased by 68% as compared to that before optimization. The yield of 6-hydroxynicotinic acid (6-HNA) produced by resting cell biotransformation reached 124.77 g/L. Accordingly, strain H9 had a promising application potential in the industrial production of 6-hydroxynicotinic acid.

Key words: nicotinic acid, 6-hydroxynicotinic acid, Pseudomonas putida, nicotinic acid hydroxylase, optimization of culture conditions

中图分类号: