从食品冷冻厂样品中筛选得到1株产脂肪酶的低温菌株KM1,经16S rRNA基因序列分析将其初步鉴定为Yersinia enterocolitica的亚种。该低温菌株分泌的胞外脂肪酶Hp-KM1经40%硫酸铵沉淀、超滤浓缩、Sephacry^TM HRS-100分子筛和Superdex G75凝胶过滤后.得到电泳纯的酶产物.纯化倍数为26倍,回收率为10.3%,相对分子质量约为34.3kD。在0-25℃和pH7.2-10范围内均保持良好的催化活性;最适催化温度为37℃,最适pH值为9.0,为典型的低温碱性脂肪酶。
KM 1 strain with cold-adapted lipase activity was isolated from frozen food factory. Based on the 16S rRNA gene sequence, the newly isolated strain KM1 was identified as a subspecies of Yersinia enterocolitica. Purification of the extracellular lipase was carried out by using sequential 40% ammonium sulphate precipi- tation, ultrafiltration, Sephacry^TM HRS-100 gel permeation chromatography and Superdex G75 column from culture supernatant to gel electrophoretic homogeneity, resulting in a 26-fold purification with a final yield of 10.3%. The molecular weight was estimated to be 34.3 kD. Lip-KM1 was a cold-adapted lipase with high ac- tivity and stable under the conditions of low temperature of 0-25 ℃ and pH 7.2-10. The optimum temperature and pH were 37 ℃ and 9.0, respectively. So it was a typical cold-adaptive alkaline lipase.