野生型天然rhlC基因缺失菌株铜绿假单胞A39-1的鼠李糖脂产物结构与合成路径解析  

Structure and synthetic pathway analysis of rhamnolipid products from wild-type natural rhlC gene deletion strain Pseudomonas aeruginosa A39-1

作　　者：罗惠芳 戚建华 童文涛 王瑞 吕天翊 宋增福[1,2,3,4] LUO Huifang;QI Jianhua;TONG Wentao;WANG Rui;LYU Tianyi;SONG Zengfu(College of Fisheries and Life Science,Shanghai Ocean University,Shanghai 201306,China;National Demonstration Center for Experimental Fisheries Science Education,Shanghai Ocean University,Shanghai 201306,China;Key Laboratory of Exploration and Utilisation of Aquatic Genetic Resources Ministry of Education,Shanghai Ocean University,Shanghai 201306,China;China-ASEAN Belt and Road Joint Laboratory on Mariculture Technology,Shanghai Ocean University,Shanghai 201306,China)

机构地区：[1]上海海洋大学水产与生命学院,上海201306 [2]上海海洋大学水产科学国家级实验教学示范中心,上海201306 [3]上海海洋大学水产种质资源发掘与利用教育部重点实验室,上海201306 [4]中国-东盟海水养殖技术“一带一路”联合实验室,上海201306

出　　处：《上海海洋大学学报》2025年第1期111-122,共12页Journal of Shanghai Ocean University

基　　金：上海市协同创新中心基金项目(ZF1206);国家重点研发计划(2022YFE0203900)。

摘　　要：为了探究铜绿假单胞菌(Pseudomonas aeruginosa)A39-1菌株以不同碳源发酵所得鼠李糖脂抑菌活性及结构组成,解析铜绿假单胞菌A39-1鼠李糖脂生物合成路径。利用牛津杯法检测鼠李糖脂对常见病原菌的抑菌活性,采用液相色谱-质谱联用技术(LC-MS)检测与分析鼠李糖脂样品的同系物组成,通过第二代测序技术(Illumina Hiseq)和第三代单分子测序技术(Pac Bio)测定和组装铜绿假单胞菌A39-1的全基因组序列,注释分析鼠李糖脂合成相关基因。结果显示,菌株A39-1以甘油、葡萄糖、葵花籽油、菜籽油和调和油作为唯一碳源发酵生产的鼠李糖脂具有抑菌活性;LC-MS检测为4种单糖单脂(Rha-C_(8∶2)、Rha-C_(9∶2)、Rha-C_(11∶2)和Rha-C_(10))和4种单糖双脂(Rha-C_(6∶1)-C_(6∶1)、Rha-C_(12∶1)-C_(12∶1)、Rha-C_(12)-C_(14)和Rha-C_(14∶2)-C_(14∶2)),Rha-C_(11:2)为主要结构;基因组预测到alg C、rml A、rml B、rml C、rml D、rhl A、rhl B相关鼠李糖脂合成基因,缺失编码合成双鼠李糖脂的鼠李糖基转移酶Ⅱ基因rhl C。研究表明,铜绿假单胞菌A39-1为野生型天然rhl C基因缺失株,以不同碳源发酵均能专一性产生单鼠李糖脂,且该单鼠李糖脂具有抑菌活性。本研究探索单鼠李糖脂抑菌活性,解析其组成结构及生物合成路径,为开发水生动物疾病新型绿色防控药物提供新思路。To explore the antibacterial activity and structural composition of rhamnolipids produced by Pseudomonas aeruginosa A39-1 fermentation with various carbon sources,as well as to analyze the rhamnolipid biosynthesis process,the inhibitory activity of rhamnolipids against pathogens was determined using the Oxford cup assay,the congener composition of rhamnolipid samples was detected and analysed using liquid chromatography-mass spectrometry(LC-MS),and the whole genome sequence was determined and assembled using Illumina Hiseq and PacBio.Genome sequence annotated and analysed rhamnolipid synthesis-related genes.Rhamnolipids produced by strain A39-1 by fermentation with glycerol,glucose,sunflower oil,rapeseed oil and blended oil as the sole carbon source had antibacterial activity;LC-MS detection of rhamnolipids obtained by glycerol fermentation as four monosaccharide monolipids(Rha-C_(8∶2),Rha-C_(9∶2),Rha-C_(11∶2),Rha-C_(10))and four monosaccharide bilipids(Rha-C_(6∶1)-C_(6∶1),Rha-C_(12∶1)-C_(12∶1),Rha-C_(12)-C_(14),Rha-C_(14∶2)-C_(14∶2)),Rha-C_(11∶2) is the primary structure;The genome was anticipated to have genes for rhamnolipid synthesis related to algC,rmlA,rmlB,rmlC,rmlD,rhlA,rhlB,as well as the absence of gene rhlC encoding rhamnosyltransferase II synthesizes bi-rhamnolipids.Pseudomonas aeruginosa A39-1,a wild-type strain with a natural rhlC gene deletion,produced monorhamnolipids exclusively by fermentation with different carbon sources,and the mono-rhamnolipids had antibacterial activity.This study explores the antibacterial activity of mono-rhamnolipids,analyzes composition and biosynthesis pathways,and provides new ideas for the development of new green prevention and control medicines for aquatic animal diseases.

关 键 词：铜绿假单胞菌A39-1 鼠李糖脂 抑菌活性 液相色谱-质谱联用技术 全基因测序分析 

分 类 号：S917.1[农业科学—水产科学]