研究报告
中国酿造
2017年第36卷第5期
总第303期
紫色红曲霉FBKL3.0018产酯化酶的酶学性质研究
胡娜12，吴鑫颖2*李付丽12王艳2邱树毅12，吴海
·123-
（1.贵州大学贵州省发酵工程与生物制药重点实验室贵州贵阳5500252.贵州大学酿酒与食品工程学院贵州贵阳550025：
3.青酒集团有限公司贵州黔东南556000）
摘要以从紫色红曲霉FBKL3.0018中固态发醇酵得到的粗酶制剂为研究对象对该菌株的酯化酶酶学性质进行了较系统的研究。通过单因素实验确定该酶最适温度为40℃在30C条件下稳定性较好最适pH值为6.5在pH5.5～7.5条件下稳定性好。Fe*、Na*、Mn* Mg、Lit、Zn"对醋化酶酶活力有抑制作用Ca在低浓度时有促进作用高浓度时有抑制作用。K*对醋化酶酶活力有促进作用并且随着K+浓度的升高，促进作用逐渐增强。吐温-80、十二烷基硫酸钠(SDS)、聚乙二醇6000、阿拉伯胶4种表面活性剂对酯化醇酶活力都有抑制作用。另外，甲醇和乙醇对该酯化酶酶活力有抑制作用，甲酸、乙酸、乳酸对酯化酶酶活力有促进作用。乙酸乙酯在低浓度时
0.015 4 mol/L.
关键词紫色红曲霉固态发酵醋酯化酶酶学性质
中图分类号TS261.1
文章编号0254-5071(2017)05-0123-05
doi:10.11882/j.issn.02545071.2017.05.026
Enzymatic properties of esterifying enzyme from Monascus purpureus FBKL3.0018
HU Na', WU Xinying**, LI Fuli', WANG Yan'2, QIU Shuyi'-, WU Hai
(I.Guizhou Key Lab of Fementation Engincering and Biological Pharmacy, Guizhou University, Guiyang 550025, China,; 2.Schoof of Liquor and Food Engineering, Guizhou University, Guiyang 550025, China; 3.Qingjiu Group Co., Ltd., Qiandongnan 556000, China)
Abstract Using the crude enzyme obtained from Monascus purpureus FBKL3.0018 by solid-state fementation as research object, the enzymatic properties of the strain was studied systematically. By single factor experiments, the optimum temperature and pH of the enzyme was 40 °C and 6.5, respectively. The enzyme was stable at 30 C and pH 5.5-7.5. Fe*, Nat, Mn*, Mg*, Lit, and Zn* had inhibition effect on enzyme activity; Ca* showed promotion effect at low concentration, however, it showed inhibition effect at high concentration. K* showed promotion effect on enzyme activity, and with the increase of K* concentration, the promotion effect gradually enhanced. Tween-80, sodium dodecyl sulfate (SDS), polyethylene glycol-6000 and Arabic gum had inhibitory effeet on enzyme activity. In addition, methanol and ethanol had inhibition effect on enzyme activity, formic acid, acetic acid and lactic acid had promotion function to enzyme activity. Ethyl acetate showed inhibition effect on esterifying enzyme activity at low con-centration, but when the concentration was 30%, it showed promotion effect. The maximum reaction velocity (V..) of the esterifying enzyme was 0.016 mol/(L-min), Michaelis constant (K,.) was 0.015 4 mol/L.
Key words Monascus pupureus, solid-state fermentation; esterifying enzyme; enzymatic properties
酯化酶包括脂肪酶和酯酶酯酶羧酸酯酶carboxy-lesterases)与脂肪酶功能相似都可以催化酯类的水解与合成")。白酒生产上常用高产酯化酶的菌株以及酯化酶制剂来提高酯的合成有利于增加白酒香味成分中酯的含量2-5)。目前,关于酯化酶酶学性质的研究主要集中在酶的最适反应温度最适pH值温度稳定性酸碱稳定性金属离子、表面活性剂和有机溶剂等对活性的影响等
胡长浩等17]对金黄色酿脓葡萄球菌（Staphylococcus aureus)FL-12产耐热性脂肪酶的酶学性质进行了研究发现脂肪酶的最适温度是45℃最适pH值为10有较好的酸碱稳定性和温度稳定性。K*对该酶有激活作用Pb、Ba有抑制
收稿日期2017-01-16
作用低浓度十二烷基硫酸钠(sodiumdodecylsulfateSDS）抑制该酶的活性低浓度乙二胺四乙酸（ethylenediaminete-traacetic acid,EDTA）对该酶几乎没有影响;1%的TritonX 100和吐温-80对酶活有促进作用。胡朝阳等(对不动杆菌 GXL02产脂肪酶的酶学性质进行了研究发现酶的最适温度为50℃最适pH值为9.0，具有较好的温度稳定性和pH 稳定性。王琰等例对真菌FL002产耐酸性脂肪酶的酶学性质进行研究结果发现最适pH为6.5在pH6.0～8.5之间酶的稳定性较好40C时酶具有很好的稳定性。研究还发现 Fe2*、Ca*对脂肪酶活性产生抑制作用Cu*、Ni*抑制作用明显这跟其他脂肪酶的酶学性质研究结果不同。黑曲霉产
基金项目责州省科技合作计划项目（黔科合LH字[2014]7675号）责州省星火计划项目（黔科合成转字[2015]5337号）
作者简介胡(1993-）女硕士研究生研究方向为酶工程*通讯作者吴鑫额(1975-）女副教授硕士研究方向为酶工程
?1994-2018 China Academic Journal Electronic Publishing House, All rights reserved.
http://ww.cnki.net