橙皮素单葡萄糖苷的酶法生物合成与定向调控

  • 王幻 ,
  • 王玉涛 ,
  • 陈良 ,
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  • 1(华南理工大学 食品科学与工程学院,广东 广州,510640) 2(喀什大学 生命与地理科学学院,新疆 喀什,834000) 3(山东奔月生物科技有限公司,山东 东营,257000)
硕士研究生

收稿日期: 2018-04-13

  修回日期: 2018-05-17

  网络出版日期: 2018-11-22

基金资助

国家自然科学基金项目(U1203183);广东省科技计划项目(2014A020209019和2015A020210039)

Enzymatic biosynthesis and directional regulation of hesperetin-7-O-glucoside

  • WANG Huan ,
  • WANG Yu-tao ,
  • CHEN Liang ,
  • et al
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  • 1(School of Food Science and Engineering, South China University of Technology, Guangzhou 510640, China) 2(School of Life and Geography, Kashi University, Kashi 834000, China) 3(Shandong BenYue Bio-Technology Co.Ltd, Dongying 834000, China)

Received date: 2018-04-13

  Revised date: 2018-05-17

  Online published: 2018-11-22

摘要

采用高效液相色谱法(HPLC)测定橙皮苷酶的酶学性质,并探讨其催化橙皮苷转化为橙皮素单葡萄糖苷(hesperetin-7-O-glucoside, HMG)的反应历程与定向调控。结果表明:橙皮苷酶在20~70 ℃、pH 3.0~7.0具有较好的稳定性,且最适反应温度为60 ℃,最适反应pH值为4.0;在最适反应温度和pH下,该酶的动力学常数Km为296.69 μg/mL,最大反应速率Vm为0.872 3 μg/(mL·min);与HMG转化相关的关键酶α-L-鼠李糖苷酶的Km为232.58 μg/mL,Vm为0.397 9 μg/(mL·min);在60 ℃、pH 4.0,酶用量为15.35 U、增溶剂添加量为0.8%、葡萄糖添加量为3 mg/mL、酶解时间为3 h时,酶反应向有利于HMG转化的方向进行,此时HMG得率为35.8%。

本文引用格式

王幻 , 王玉涛 , 陈良 , . 橙皮素单葡萄糖苷的酶法生物合成与定向调控[J]. 食品与发酵工业, 2018 , 44(10) : 7 -13 . DOI: 10.13995/j.cnki.11-1802/ts.017537

Abstract

Hesperetin-7-O-glucoside (HMG) is a precursor substance of a new sweetener. The enzymatic properties of hesperidinase were determined by high performance liquid chromatography (HPLC). The reaction mechanism and directional regulation of the conversion of hesperidin to HMG were discussed. Results showed that hesperidinase had good stability at 20-70 ℃ and pH 3.0-7.0, the optimum reaction temperature and pH was 60 ℃ and 4.0, respectively. Furthermore, at the optimum reaction temperature and pH, the kinetic constant Km of hesperidinase was 296.69 μg/mL, the maximum reaction rate Vm was 0.872 3 μg/(mL·min). Also, the Km of α -L- rhamnosidase related to HMG transformation was 232.58 μg/mL and the Vm was 0.397 9 μg/(mL·min). The optimal hydrolysis conditions were pH 4.0 and hydrolysis temperature of 60 ℃, enzyme dosage 15.35 U, the amount of solubilizer 0.8%, the amount of glucose 3 mg/mL, and hydrolysis time 3 h. The enzymatic reaction was carried out using these conditions in favor of HMG transformation, and the yield of HMG was 35.8%.
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