Abstract: In the present study, a preliminary purification was carried out to obtain heat-resistant lipase secreted by Pseudomonas fluorescens separated from fresh milk. Moreover, the effects of temperature, pH, and Ca2+ concentration on the activity of this lipase were determined. Results showed that the lipase from Pseudomonas fluorescens had a molecular weight 55 kD and a purity of 91.5% after the purification, and its relative activity was 75.02%, 86.21%, 73.25%, 17.62% and 13.63% after heat treatment at 50 ℃ for 30 min, 63 ℃ for 30 min, 72 ℃ for 20 s, 90 ℃ for 10 min, and 121℃, 0.1 MPa for 20 min, respectively. The optimum reaction pH and temperature for this enzyme were 8.0 and 63 ℃, respectively. The lipase activity was not significantly affected by Ca2+ concentration.

Key words: Pseudomonas fluorescens lipase; thermostable; isolation; activity

摘要： 以新鲜牛乳中分离到的荧光假单胞菌为研究对象,对其分泌产生的耐热性脂肪酶进行纯化,并研究不同温度、pH值、Ca2+质量浓度对该脂肪酶活性的影响.结果表明:经过初步纯化的脂肪酶分子质量为55 kD,纯度为91.5％,并且发现该耐热性脂肪酶在50℃、30 min,63℃、30 min,72℃、20 s,90℃、10 min,121℃、0.1 MPa、20 min处理之后,相对酶活力分别为75.02％、86.21％、73.25％、17.62％、13.63％;并且该脂肪酶的最适温度为63℃,最适pH值为8.0;在荧光假单胞菌体外,Ca2+质量浓度对荧光假单胞菌脂肪酶的活力并无显著影响.

关键词: 荧光假单胞菌；脂肪酶；耐热性；分离；活性