Dandavate Vrushali, Jinjala Jayesh, Keharia Haresh, Madamwar Datta
BRD School of Biosciences, Sardar Patel Maidan, Sardar Patel University, Vallabh Vidyanagar, Gujarat, India.
Bioresour Technol. 2009 Jul;100(13):3374-81. doi: 10.1016/j.biortech.2009.02.011. Epub 2009 Mar 12.
Burkholderia multivorans V2 (BMV2) isolated from soil was found to produce an extracellular solvent tolerant lipase (6.477 U/mL). This lipase exhibited maximum stability in n-hexane retaining about 97.8% activity for 24h. After performing statistical optimization of medium components for lipase production, a 2.2-fold (14 U/mL) enhancement in the lipase production was observed. The crude lipase from BMV2 was partially purified by ultrafiltration and gel permeation chromatography with 24.64-fold purification. The K(m) and V(max) values for partially purified BMV2 lipase were found to be 1.56 mM and 5.62 micromoles/mg min. The metal ions Ca(2+), Mg(2+) and Mn(2+) had stimulatory effect on lipase activity, whereas Cu(2+), Fe(2+) and Zn(2+) strongly inhibited the lipase activity. EDTA and PMSF at 10mM concentration strongly inhibited the lipase activity. Non-ionic and anionic surfactants stimulated the lipase activity. BMV2 lipase was proved to be efficient in synthesis of ethyl butyrate ester under non-aqueous environment.
从土壤中分离出的多食伯克霍尔德菌V2（BMV2）被发现能产生一种胞外耐溶剂脂肪酶（6.477 U/mL）。这种脂肪酶在正己烷中表现出最大稳定性，24小时内保留约97.8%的活性。在对脂肪酶生产的培养基成分进行统计优化后，观察到脂肪酶产量提高了2.2倍（14 U/mL）。来自BMV2的粗脂肪酶通过超滤和凝胶渗透色谱法进行部分纯化，纯化倍数为24.64倍。发现部分纯化的BMV2脂肪酶的K(m)和V(max)值分别为1.56 mM和5.62微摩尔/毫克·分钟。金属离子Ca(2+)、Mg(2+)和Mn(2+)对脂肪酶活性有刺激作用，而Cu(2+)、Fe(2+)和Zn(2+)则强烈抑制脂肪酶活性。10 mM浓度的EDTA和PMSF强烈抑制脂肪酶活性。非离子和阴离子表面活性剂刺激脂肪酶活性。已证明BMV2脂肪酶在非水环境中合成丁酸乙酯酯方面是有效的。
