摘要
【目的】从自然界筛选出脂肪酶高产菌株,鉴定其种属并对其酶学性质进行研究.【方法】从白银市不同地点油污染河流和土壤中采集样品,利用维多利亚蓝培养基与中性红油脂培养基进行初筛,改进铜皂-分光光度计法测定酶活力进行复筛,再结合形态观察、生理生化鉴定和16SrDNA序列分析进行菌种鉴定,并对脂肪酶的酶学性质进行研究.【结果】分离筛选出一株产脂肪酶的细菌BY-8,鉴定为粪产碱杆菌(Alcaligenes faecalis).该菌株所产脂肪酶在40℃和pH 7.0的条件下酶活力为3.64U/mL.酶学性质研究结果表明,产生的脂肪酶最适作用温度和pH分别为35℃和7.0,Mg^(2+)和Ca^(2+)对脂肪酶活性有明显地促进作用,而Cu^(2+)和Zn^(2+)对酶活性有抑制作用,EDTA和SDS可使脂肪酶失去活力.【结论】菌株BY-8所产脂肪酶的酶活力一般,但酶学性质稳定,是一种中性脂肪酶.
【Objective】This study aimed to screen lipase high-yielding strains from nature,identify the species and research the enzymatic properties.【Method】Collecting samples from oil polluted rivers and soil at different sites in Baiyin city to conduct primary screen using Victoria blue medium and neutral red oil medium and rescreening by determining the enzyme activity using the improved copper soap-spectrophotometer method,identify the stains by morphological observation,physiological and biochemical identification,and16S rDNA sequence analysis,finally study the enzymatic properties of lipase.【Result】A bacterial strain BY-8producing lipase was isolated and identified as Alcaligenes faecalis with lipase activity of3.64U/mL under the condition of40℃and pH7.0.The optimal reaction temperature and pH for producing lipase from BY-8was35℃and7.0,respectively.The lipase activity of BY-8was promoted by Mg2+and Ca2+and inhibited by Cu2+and Zn2+,and appeared inactivation after being treated by EDTA and SDS.【Conclusion】The lipase produced by BY-8strain was a neutral lipase with ordinary activity but stable enzymatic properties.
作者
刘晓丽
刘玉华
韩生义
马才
杨孝朴
LIU Xiao-li;LIU Yu-hua;HAN Sheng-yi;MA Cai;YANG Xiao-pu(College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China;Henan Suokang Biotechonology,Co.Ltd,Jiaozuo 454950,China)
出处
《甘肃农业大学学报》
CAS
CSCD
北大核心
2018年第1期22-28,共7页
Journal of Gansu Agricultural University
基金
甘肃省财政厅"微生物脂肪酶基因克隆与表达研究"项目(2013NY-18)
关键词
脂肪酶
筛选
鉴定
酶学性质
lipase
screening
identification
enzymatic properties
作者简介
通信作者:杨孝朴,男,教授,硕士生导师,研究方向为生物化学与分子生物学.E-mail:yangxpu@gsau.edu.cn
