南极Shewanella sp．NJ49低温降解酶的定域及特性研究  

Localization of Antarctic Degradation Enzyme Shewanella sp. N J49 Bacteria and Characteristics Study

作　　者：王以斌[1,2] 刘晋 刘芳明[1,2] 缪锦来[1,2] 郑洲[1,2] 单连菊 

机构地区：[1]国家海洋局第一海洋研究所,山东青岛266061 [2]国家海洋局海洋生物活性物质重点实验室,山东青岛266061 [3]青岛华仁药业股份有限公司,山东青岛266101 [4]青岛啤酒股份有限公司,山东青岛266071

出　　处：《海洋科学进展》2011年第A01期43-48,共6页Advances in Marine Science

基　　金：国家自然科学基金--南极海冰区石油烃低温降解微生物及其低温降解机制研究（40876107）;国家海洋局青年海洋科学基金--南极嗜冷菌烷烃羟化酶基因的克隆及其结构特征研究（2010122）

摘　　要：石油烃的微生物降解过程与微生物细胞所包含的酶及其酶促反应有关，本文以南极低温降解菌Shewanellasp．NJ49为研究对象，采用2216E和MMC人工海水培养基培养，利用静息细胞技术提取NJ49的胞外酶、膜周酶及膜内酶，研究三种酶对正十六烷的低温降解能力、代谢产物分析，测定不同分子量区段酶的活性。结果表明：胞外酶对正十六烷的降解率为63％，膜周酶的降解率小于38％，膜内酶无降解能力，Shewanellasp．NJ49低温降解酶主要定域为胞外酶和膜周酶；酶代谢过程中生成十六醛、十六醇为主的产物，酶以末端氧化方式完成初始降解；烷烃氧化的关键酶（羟化酶）集中在分子量为3万～10万的区段。Shewanella sp. NJ49 was cultured with 2216E and manual seawater for enzyme study which play key role in procession of degradation of petroleum hydrocarbon. Extracellular enzyme, periplasmic and cytoplasmic fractions were extracted after osmotic shock. Efficacy of hexadecane degradation was compared through three enzyme and metabolize product was analyzed by GC and GC-MS . Four enzymes of different molecular weight （greater than 100 000, 30 000;100 000, 10 000;30 000 and 5 000;10 000, respectively） were collected by ultrafiltration and then determined and compared the enzymatic activity. The results showed that degradation rate of extracellular enzyme and periplasmic enzyme was above 60 % and less than 40%,respectively, but capacity of degradation hexadecane of cytoplasmic enzyme was zero. Degradation enzyme of Shewanella sp. NJ49 was located extracellular and periplasmic emzyme; Hexadecanal and 1- hexadecanol were the main products in the process of n-hexadecane degradation, It accomplished the pre- liminary degradation by the terminal oxidation. Relative molecular mass of Monooxygenase was 0.03;0.1 million.

关 键 词：SHEWANELLA sp.NJ49 正十六烷 定域 胞外酶 

分 类 号：Q936[生物学—微生物学]