拟青霉FLH30 β-葡萄糖苷酶动力学、激活抑制及其转糖苷

华承伟; 于江傲; 李兰; 常景玲; 张志宏

doi:10.13386/j.issn1002-0306.2016.16.018

拟青霉FLH30 β-葡萄糖苷酶动力学、激活抑制及其转糖苷

河南科技学院生命科技学院

基金项目:

河南科技学院重点科研项目(201010611004)；

详细信息

作者简介:
华承伟(1972-),男,博士,副教授,研究方向:酶工程、食品生物技术,E-mail:cwhua@hist.edu.cn。;

中图分类号: TS201.2
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出版历程
- 收稿日期: 2015-07-05

Kinetics,activation,inhibition and transglycosylation of the β-glucosidase from Paecilomyces sp.FLH30

School of Life Science and Technology,Henan Institute of Science and Technology

摘要

摘要: 研究了拟青霉FLH30胞内β-葡萄糖苷酶动力学性质及葡萄糖、木糖和葡萄糖醛酸内酯对该酶的激活及抑制作用。结果表明:该酶对p NP-β-D-葡萄糖苷、p NP-β-D-半乳糖苷、纤维二糖、乳糖、龙胆二糖和水扬苷的水解动力学常数Km分别为0.54、5.28、0.98、26.34、6.92和1.72 mmol/L;在以对硝基苯基-α-D-吡喃葡萄糖苷（p NPG）为底物时,5¹50 mmol/L葡萄糖和20⁶00 mmol/L木糖对酶有激活作用,高于此范围,则表现出抑制作用,葡萄糖醛酸内酯具有很强的抑制作用,葡萄糖、木糖和葡萄糖醛酸内酯抑制常数Ki值分别为63.4、170.3和0.038 mmol/L。木糖能促进该酶对纤维二糖和乳糖的水解作用,木糖浓度为200 mmol/L时,酶活分别增加189.6%和166.3%。该酶还具有转糖苷活性,能够将纤维二糖部分转化为纤维三糖和龙胆二糖。
- β-葡萄糖苷酶 /
- 动力学 /
- 激活 /
- 抑制 /
- 转糖苷
Abstract: The β- glucosidase from Paecilomyces sp. FLH30 exhibited a broad substrate specificity. The enzyme hydrolyzed p NPG,p NP- β- D- galactoside,cellobiose,lactose,salicin,gentiobiose,exhibiting apparent kinetics constant( Km) values of 0.54,5.28,0.98,26.34,6.92 and 1.72 mmol/L,respectively.A certain concentration of glucose( 5~150 mmol/L) and xylose( 20~600 mmol/L) had an activation on the enzymes use p NPG as substrate,above the value then demonstrated inhibition.Glucuronolactone had strongly inhibited the enzyme activity,the value of inhibite constant Kiof glucose,xylose and glucuronolactone was 63.4,170.3 and 0.038 mmol / L. Xylose also had an activation on the enzyme hydrolysis of cellobiose and lactose,the enzyme activity increased by 189.6% and166.3% respectively in the xylose concentration 200 mmol / L. Furthermore,the enzyme showed transglycosylation activity and could produced cellotriose and gentibiose use cellobiose as substrate.
- β-glucosidase /
- kinetics /
- activation /
- inhibition /
- transglycosylation

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