嗜盐脂肪酶产生菌的筛选及其粗酶性质.docVIP

嗜盐脂肪酶产生菌的筛选及其粗酶性质 张 萌1,2 张晓梅1 窦文芳1 许泓瑜1 许正宏1,2* (1. 江南大学医药学院制药工程研究室 无锡 214122) (2江南大学工业生物技术教育部重点实验室 无锡 214122)从内蒙古锡林浩特地区盐湖一株嗜盐脂肪酶产菌, 鉴定Haloterrigena thermotolerans Z4。该菌最适生长NaCl浓度为3.5 mol/L60(C, 属于嗜盐耐热古生菌。Ba2+、Fe2+Cu2+)对酶有激活作用EDTA的抑制PMSF的完全抑制NaCl有较高的依赖专一性, 至少需要0.5 mol/L NaCl维持活性并且高浓度NaCl可以提高耐热水平p-NPB)的最适水解条件为：pH 8.0、70(C、3.5 mol/L NaCl; 而对长链底物对硝基苯酚十六酸酯(p-NPP)的最适水解条件为：pH 8.0、80(C、2.5 mol/L NaCl。 关键词: 嗜盐脂肪酶, 嗜盐古生菌, 鉴定, 酶学性质 Screening of a Halobacteria Strain Producing Halophilic Lipase and Characterization of Crude Enzyme ZHANG Meng1,2 ZHANG Xiao-Mei1 DOU Wen-Fang1  XU Hong-Yu1 XU Zheng-Hong1,2* (1. Laboratory of Pharmaceutical Engineering, School of Medicine and Pharmaceutics, Jiangnan University, Wuxi, Jiangsu 214122, China) (2. The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu 214122, China) Abstract: A halobacteria strain Z4 producing extracellular halophilic lipase screened from Hypersaline lakes of Inner Mongolia was identified as Haloterrigena thermotolerans. The crude lipase from Z4 was partially characterized using p-NPB. The activity of crude lipase was markedly increased 20%~30% by metal ions (Ba2+,Fe2+,Cu2+), but was obviously decreased 20% by EDTA. The lipase was completely inhibited by PMSF. The activity and heat resistance of enzyme was increased by NaCl, suggesting that the enzyme has strong dependence and specificity on NaCl. The thermostability of the crude enzyme was increased by alcohols, the glycerin is the best one. The optimum conditions for the crude lipase hydrolyzing p-NPB were followed: 3.5 mol/L NaCl, 70(C and pH 8.0, and the optimum conditions for the crude lipase hydrolyzing p-NPP were followed: 2.5 mol/L NaCl, 80(C and pH 8.0. Keywords: Halophilic lipase, Halophilic archaeon, Identification, Characterization随着工业生物技术的不断发展, 脂肪酶在工业生物技术领域的应用逐渐成为研究的热点, 脂肪酶广泛应用于洗涤剂、食品、医药、制革、纺织及废物处理等领域[1]。脂肪酶按其作用特点主要可以分为碱性脂肪酶、酸性脂肪酶耐热脂肪酶低温脂肪酶, 而来自嗜盐古生菌的嗜盐脂肪酶却鲜见报道。嗜盐古生菌是生长在盐湖、盐碱湖、盐沼、死海和盐场等高盐