一株产胞外多糖海洋弧菌的分离鉴定及其多糖抗肿瘤活性初步研究

生物技术通报 ›› 2016, Vol. 32 ›› Issue (12): 166-171.

一株产胞外多糖海洋弧菌的分离鉴定及其多糖抗肿瘤活性初步研究

龙寒¹,陈盛峰¹,陈佳¹,杨迪¹,李晓燕¹,黄玉油¹,何秀苗¹，²,禤金彩¹

1. 广西民族大学海洋与生物技术学院广西高校微生物与植物资源利用重点实验室，南宁 530006；
2. 广西民族大学广西林产化学与工程重点实验室，南宁 530006

收稿日期:2016-05-30 出版日期:2016-12-25 发布日期:2016-12-07
作者简介:龙寒，女，实验师，研究方向：海洋微生物学；E-mail：longhoo@163.com
基金资助:
国家自然科学基金项目（31560706），2014广西民族大学相思湖青年学者创新团队资助，广西林产化学与工程协同创新高水平研究项目（2013B01）

Isolation and Identification of Marine Vibrio Producing Extracellular Polysaccharide（EPS）and the Preliminary Study on Anti-tumor Activity of the EPS

LONG Han¹,CHEN Sheng-feng¹,CHEN Jia¹,YANG Di¹,LI Xiao-yan¹,HUANG Yu-you¹,HE Xiu-miao¹，²,XUAN Jin-cai¹

1. School of Marine Sciences and Biotechnology/Guangxi Colleges and Universities Key Laboratory of Utilization of Microbial and Botanical Resources，Guangxi University for Nationalities，Nanning 530006；
2. Guangxi Key Laboratory of Chemistry and Engineering of Forest Products，Nanning 530006

Received:2016-05-30 Published:2016-12-25 Online:2016-12-07

摘要/Abstract

摘要： 为初步探究广西北部湾海洋细菌所产胞外多糖（Extracellular polysaccharide，EPS）的生物学活性，通过LB-苯胺蓝培养基分离产EPS的海洋细菌，苯酚-硫酸法测定EPS产量，16S rDNA法鉴定多糖产量较高的菌株，通过凝胶渗透色谱测定多糖分子量，红外光谱分析该多糖的结构；最后运用MTT（噻唑蓝）法测定该EPS对Vero细胞的毒性，以及对HeLa细胞的抑制作用。结果显示，从海水及红树林泥样中共分离获得167株产EPS的海洋细菌，其中一株BHc-09 EPS产量较高，达0.306 mg/mL，通过16S rDNA序列分析鉴定BHc-09为弧菌属，其EPS分子量约为184.5 kD，结构中含有糖醛酸；该胞外多糖对正常Vero细胞无毒性而对肿瘤细胞的生长具有明显的抑制作用。

关键词: 海洋弧菌, 胞外多糖, 分离鉴定, 抗肿瘤

Abstract: In order to investigate the bio-activities of the extracellular polysaccharide（EPS）from the marine bacteria in the Guangxi Beibu Gulf，the marine bacteria of EPS-producing were isolated from water/mud samples by LB-aniline blue medium. Phenol-sulfuric acid method was used for measuring the yield of the EPS. The marine bacterial strains with high-yield of ESP were further identified by the sequence analysis of 16S rDNA. The molecular weight and the preliminary structure of the EPS were tested by Gel permeation chromatography（GPC）and infrared spectrometry，respectively. Finally，the cytotoxicity of the EPS on Vero cells and the inhibitory effect of the EPS on Hela cells were tested by the MTT method. The results showed that totally 167 marine bacteria producing EPS were isolated from sea water/mud of mangrove，one of which（named BHc-09）yielded ESP as high as 0.306 mg/mL and was identified to be Vibrio by the analysis of its 16S rDNA sequence. The molecular weight of the EPS from BHc-09 was 184.5 kD，and there was uronic acid in the EPS structure. The EPS produced by BHc-09 presented no cytotoxicity on Vero cell，but significantly inhibitory effects on tumor cells. The results suggested that an EPS-producing marine Vibrio was successfully isolated and identified，and its extracellular polysaccharide has anti-tumor activity.

Key words: marine Vibrio, extracellular polysaccharide, isolation and identification, anti-tumor

中图分类号:

10.13560/j.cnki.biotech.bull.1985.2016.12.026

龙寒,陈盛峰,陈佳,杨迪,李晓燕,黄玉油,何秀苗，禤金彩. 一株产胞外多糖海洋弧菌的分离鉴定及其多糖抗肿瘤活性初步研究[J]. 生物技术通报, 2016, 32(12): 166-171.

LONG Han,CHEN Sheng-feng,CHEN Jia,YANG Di,LI Xiao-yan,HUANG Yu-you,HE Xiu-miao，XUAN Jin-cai. Isolation and Identification of Marine Vibrio Producing Extracellular Polysaccharide（EPS）and the Preliminary Study on Anti-tumor Activity of the EPS[J]. Biotechnology Bulletin, 2016, 32(12): 166-171.

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