深海沉积物宏基因组中新型对羟基苯丙酮酸双加氧酶基因的鉴定  

Identification of novel 4-hydroxyphenylpyruvate dioxygenase genes from the deep sea sediment metagenomes

作　　者：张欢 吴科锋 刘景川 汤熙翔 ZHANG Huan;WU Kefeng;LIU Jingchuan;TANG Xixiang(Marine Biomedical Research Institution,Guangdong Medical University,Zhanjiang 524023,China;Third Institute of Oceanography,MNR,Xiamen 361005,China;Guangdong Provincial Laboratory of Southern Marine Science and Engineering,Zhanjiang 524023,China;Longyan Tobacco Industry Limited Liability Company,Longyan 364000,China)

机构地区：[1]广东医科大学海洋医药研究院,广东湛江524023 [2]自然资源部第三海洋研究所,福建厦门361005 [3]南方海洋科学与工程广东省实验室(湛江),广东湛江524023 [4]龙岩烟草工业有限责任公司,福建龙岩364000

出　　处：《应用海洋学学报》2024年第3期442-450,共9页Journal of Applied Oceanography

基　　金：亚洲海洋合作基金(99950410);福建省科技计划项目(2021N0028)。

摘　　要：为采用不依赖培养的方法获取深海沉积物微生物遗传资源,利用深海沉积物DNA构建了2个宏基因组文库,对对羟基苯丙酮酸双加氧酶(4HPPD)活性进行了筛选,并分离出3个克隆子。体外转座子突变和全长测序发现了两个新的4 HPPD基因和一个新的尿黑酸(HGA)代谢基因簇。克隆到pMD18-T载体中的3个新的4 HPPD基因能够在大肠杆菌中产生水溶性棕色素。该色素被鉴定为黑色素,并与HGA相关。结果表明,深海沉积物宏基因组含有多样性丰富的4 HPPD功能基因,可用于发现新的黑色素生物合成基因。To access the microbial genetic resources of deep-sea sediments by a culture-independent approach,two metagenomic libraries constructed with deep sea sediments DNA were screened for the 4-hydroxyphenylpyruvate dioxygenase(4HPPD)activity and three positive clones were obtained.In vitro transposon mutagenesis and full-length sequencing revealed two novel 4 HPPD genes and a novel homogentistic acid(HGA)metabolism gene cluster.Three new 4 HPPD genes cloned into pMD18-T vector could produce water-soluble brown pigment in Escherichia coli.The pigment was identified to be pyomelanin and associated with HGA.Results show that the metagenomes of deep-sea sediments contain diverse 4 HPPD functional genes,which can be used to find new melanin biosysthesis genes.

关 键 词：海洋生物学 对羟基苯丙酮酸双加氧酶基因 尿黑酸 宏基因组文库 深海 

分 类 号：P735[天文地球—海洋生物学]