Karuppiah Valliappan, Li Yingxin, Sun Wei, Feng Guofang, Li Zhiyong
Marine Biotechnology Laboratory, State Key Laboratory of Microbial Metabolism and School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240, People's Republic of China.
Appl Microbiol Biotechnol. 2015 Jul;99(14):5939-50. doi: 10.1007/s00253-015-6547-8. Epub 2015 Mar 29.
Phenazines represent a large group of nitrogen-containing heterocyclic compounds produced by the diverse group of bacteria including actinobacteria. In this study, a total of 197 actinobacterial strains were isolated from seven different marine sponge species in the South China Sea using five different culture media. Eighty-seven morphologically different actinobacterial strains were selected and grouped into 13 genera, including Actinoalloteichus, Kocuria, Micrococcus, Micromonospora, Mycobacterium, Nocardiopsis, Prauserella, Rhodococcus, Saccharopolyspora, Salinispora, Serinicoccus, and Streptomyces by the phylogenetic analysis of 16S rRNA gene. Based on the screening of phzE genes, ten strains, including five Streptomyces, two Nocardiopsis, one Salinispora, one Micrococcus, and one Serinicoccus were found to be potential for phenazine production. The level of phzE gene expression was highly expressed in Nocardiopsis sp. 13-33-15, 13-12-13, and Serinicoccus sp. 13-12-4 on the fifth day of fermentation. Finally, 1,6-dihydroxy phenazine (1) from Nocardiopsis sp. 13-33-15 and 13-12-13, and 1,6-dimethoxy phenazine (2) from Nocardiopsis sp. 13-33-15 were isolated and identified successfully based on ESI-MS and NMR analysis. The compounds 1 and 2 showed antibacterial activity against Bacillus mycoides SJ14, Staphylococcus aureus SJ51, Escherichia coli SJ42, and Micrococcus luteus SJ47. This study suggests that the integrated approach of gene screening and chemical analysis is an effective strategy to find the target compounds and lays the basis for the production of phenazine from the sponge-associated actinobacteria.
吩嗪是一大类由包括放线菌在内的多种细菌产生的含氮杂环化合物。在本研究中，使用五种不同的培养基从中国南海的七种不同海洋海绵物种中总共分离出197株放线菌菌株。选择了87株形态不同的放线菌菌株，并通过16S rRNA基因的系统发育分析将其分为13个属，包括放线别氏菌属、考克氏菌属、微球菌属、小单孢菌属、分枝杆菌属、诺卡氏放线菌属、普劳斯氏菌属、红球菌属、糖多孢菌属、盐孢菌属、丝孢球菌属和链霉菌属。基于对phzE基因的筛选，发现十株菌株具有产生吩嗪的潜力，其中包括五株链霉菌、两株诺卡氏放线菌、一株盐孢菌、一株微球菌和一株丝孢球菌。phzE基因表达水平在发酵第五天的诺卡氏放线菌13-33-15、13-12-13和丝孢球菌13-12-4中高度表达。最后，基于电喷雾电离质谱（ESI-MS）和核磁共振（NMR）分析，成功分离并鉴定了来自诺卡氏放线菌13-33-15和13-12-13的1,6-二羟基吩嗪（1）以及来自诺卡氏放线菌13-33-15的1,6-二甲氧基吩嗪（2）。化合物1和2对蕈状芽孢杆菌SJ14、金黄色葡萄球菌SJ51、大肠杆菌SJ42和藤黄微球菌SJ47显示出抗菌活性。本研究表明，基因筛选和化学分析相结合的方法是寻找目标化合物的有效策略，并为从海绵相关放线菌生产吩嗪奠定了基础。
