Objective To separate out and identify a high yield strain for acid lipase production. Methods A strain of acid lipase was obtained by preliminary screening with plates containing the culture medium of the bromocresol purple indicator, and secondary screening by the transparent circle method with glycerin triglyceride butyrate. The enzyme activity of the screened strain was determined by the methods of plate transparent ring, olive oil emulsion titration and the method of colorimetry. Results A strain of high yield acid lipase was isolated and named WY19, and its crude enzyme activity reached 11000 U/L. Through the morphological identification of the strains WY19, identification, physiological and biochemical experiments and molecular biology combined with the phylogenetic tree analysis, the high-producing strain WY19 was determined as Klebsiella. Conclusion The lipase of strain WY19 is an acid lipase, and it has good application prospects in the fields of food, medicine, oil processing.%目的 分离筛选出一株酸性脂肪酶高产菌株并对其进行鉴定.方法 通过溴甲酚紫酸碱指示剂进行平板初筛,甘油三丁酸酯透明圈法以及摇瓶培养复筛,筛选获得了一株酸性脂肪酶产生菌株;利用平板透明圈法、橄榄油乳化液滴定法和对硝基苯酚比色法3种方法对所筛菌株进行酶活力测定.结果 分离获得一株具有高产酸性脂肪酶活力的菌株,将其命名为菌株WY19.经过检测,其粗酶活力达到11000 U/L.通过对菌株WY19进行形态学鉴定、生理生化实验以及分子生物学鉴定,结合系统发育树分析,确定本研究获得的酸性脂肪酶产生菌为克雷伯氏菌.结论 菌株WY19所产脂肪酶为酸性脂肪酶,在食品、医药、油脂加工等领域方面具有较好的应用前景.
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