盐渍海带盐中可培养嗜盐古菌多样性、胞外酶活及拮抗作用探究  

Diversity, Extracellular Enzyme Activity and Antagonism of Culturable Halophilic Archaea in Salt from Saline Kelp

作　　者：苏亦鸣 陈飞龙 张申奥 陈绍兴 SU Yi-ming;CHEN Fei-long;ZHANG Shen-ao;CHEN Shao-xing(Coll.of Life Sci.,Anhui Normal Uni.,Wuhu 241000)

机构地区：[1]安徽师范大学生命科学学院,安徽芜湖241000

出　　处：《微生物学杂志》2022年第5期19-30,共12页Journal of Microbiology

基　　金：国家自然科学基金项目(31460003);安徽省重要生物资源保护与利用开放课题项目(591601)。

摘　　要：盐渍海带盐是一种高盐环境,其中可能生存有嗜盐微生物,如嗜盐古菌。嗜盐古菌是一类生活于高盐环境的极端环境微生物。为了探究盐渍海带盐中嗜盐微生物的物种多样性,筛查分离了菌株的几种常见胞外功能酶活性和拮抗活性。采用纯培养技术,从盐渍海带盐样品中分离培养嗜盐微生物,并对其16S rRNA基因进行扩增和测序;基于16S rRNA基因序列的相似度分析,确定分离菌株在属级水平的分类地位;依据种属信息,挑选代表菌株,进行胞外常见功能酶活性和拮抗作用测定。从盐渍海带盐样品中分离到来自Haloarcula(盐盒菌属)、Halorubrum(盐红菌属)、Halarchaeum、Halobacterium(盐杆菌属)、Halococcus(盐球菌属)、Halolamina(盐薄片形菌属)和Haloplanus(盐扁平菌属)等7个属的131株嗜盐古菌;检测到产胞外蛋白酶菌株1株,产酯酶菌2株,产明胶酶菌7株,产氧化酶菌1株和产触菌酶5株;此外,筛选到6株具有拮抗活性的菌株,其中来自盐红菌属菌株Halorubrum sp. ZSA68较其他菌株生长快,产抑菌活性物质快,并显示出较强的抑菌活性和较广的抑菌谱,初步推测Halorubrum sp. ZSA68所产物质为多肽或蛋白类抑菌活性物质,其分子量大小约为30~50 kDa,该抑菌活性物质在60℃以上或NaCl浓度低于2%时容易失去活性。通过本研究认识了海带盐中可培养嗜盐古菌的分布,获得多株具有较高胞外酶活性及具拮抗作用的菌株,积累了丰富的嗜盐古菌菌株资源,这在高盐食品的防腐方面具有潜在的应用价值。Salt from saline kelp is one type of hypersaline environments, in which halophilic microorganisms, such as halophilic archaea, may exist. Halophilic archaea is a kind of extremophiles living in hypersaline environments. In order to explore the microbial community composition of halophilic microorganisms in saline kelp salt, culture-dependent approach was employed. Several extracellular enzyme activities and antagonistic activities of representatives were detected and screened from isolated strains, to provide a foundation for better excavation of the application prospect and research value of halophilic microorganisms. Pure cultivation technology was adopted to isolate and cultivated halophilic microbes, their 16S rRNA genes were amplified and sequenced;based on the similarity analysis of the 16S rRNA gene sequences, the taxonomic status of the isolated strains at genus level was determined;and according to their specific and genus information, representative strains were selected to determine their functional enzymes activity and antagonistic effects. In this study, 131 strains halophilic archaea from samples of salt of salinized kelp were isolated, they belonged to seven genera, Haloarcula, Halorubrum, Halarchaeum, Halobacterium, Halococcus, Halolamina and Haloplanus. Among them, 1 strain produced extracellular proteases, 2 strains esterase, 7 strains gelatinase, 1 strain oxidase, and 5 strains catalase were detected. In addition, 6 haloarchaeal strains with antagonistic activities were detected among them Halorubrum sp. ZSA68 from the genus of Halorubrum grew faster and produced antimicrobial active substances faster than other strains, and showed stronger antimicrobial activity and wider antibacterial spectrum than others. It has been preliminarily conjectured that Halorubrum sp. ZSA68 produced a polypeptide or protein antimicrobial substance with molecular weight of about 30-50 kDa. The antimicrobial active substance will easily lose its activity when the temperature above 60 ℃, or the concentration of NaC

关 键 词：海带盐 嗜盐古菌 物种多样性 胞外酶 抑菌活性 

分 类 号：Q935[生物学—微生物学]