耐有机溶剂脂肪酶产生菌的筛选及其粗酶酶学性质.PDFVIP

116 2012, Vol. 33, No. 03 食品科学 ※生物工程 1 1 2 1 1 李俊峰 ，李红芳 ，段效辉 ，门晋名 ，宿 烽 (1. 青岛科技大学化工学院，山东 青岛 266042 ；2. 烟台出入境检验检疫局，山东 烟台 264000) 以体积分数2% 苯为筛选压力，利用罗丹明 B 平板显色法和摇瓶发酵法，从采集的花生地土壤样品中分离 筛选得到 1 株中度耐热、耐碱脂肪酶产生菌，编号为 H2 。通过形态观察、生理生化特性实验及其 16S rDNA 基 因序列对菌种进行鉴定。结果表明，H2 菌株与短小芽孢杆菌(Bacillus pumilus) 的亲缘关系最紧密。通过研究得到 该菌株的摇瓶发酵条件：产酶培养基为：蛋白胨 3% 、酵母膏 1% 、NaCl 0.5% 、橄榄油 1% ，pH7.0 ，摇瓶发 酵温度为 28 ℃，摇床转速为 180r/min，发酵周期为48 ～60h 。所产脂肪酶在40 ℃、pH9.0 时酶活性最高，对pH 值和温度的适应范围较宽，pH6.0 ～10.0 比较稳定，35 ～50 ℃具有较高酶活性。 耐有机溶剂；短小芽孢杆菌；脂肪酶；筛选 Screening of Organic Solvent Tolerant Lipase-Producing Bacteria and Enzymatic Properties of Crude Lipase 1 1 2 1 1 LI Jun-feng ，LI Hong-fang ，DUAN Xiao-hui ，MEN Jin-ming ，SU Feng (1. College of Chemical Engineering, Qingdao University of Science and Technology, Qingdao 266042, China； 2. Yantai Entry-Exit Inspection and Quarantine Bureau, Yantai 264000, China) Abstract ：A moderately thermostable and alkali-resistant lipase-producing strain named as H2 was isolated from peanut plantation soil using 2% benzene by Rhodamine B plate assay and shake flask fermentation. It was preliminarily identified based on morphological and physiological characteristics. Meanwhile, the 16S rDNA gene of the strain was cloned and sequenced. The physiological and biochemical characterization and phylogenetic tree based on the 16S rDNA sequence showed that the strain was highly homologous to Bacillus pumilus. The optimal culture medium for lipase production by the strain was composed of 3% peptone, 1% yeast extract, 0.5% NaCl, 1% ol