物理化学学报 >> 1999, Vol. 15 >> Issue (10): 930-937.doi: 10.3866/PKU.WHXB19991013

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卵磷脂反胶束体系中β-1，4葡萄糖苷酶的研究

缪炜, 姚松年   

  1. 武汉大学测试中心，武汉 430072
  • 收稿日期:1998-10-22 修回日期:1998-12-15 发布日期:1999-10-15
  • 通讯作者:

Study of β-1，4-glucosidase in Water/PC/Heptane-Butanol Reverse Micelles

Miao Wei, Yao Song-Nian   

  1. Center of Analysis and Measurement Wuhan University,Wuhan 430072
  • Received:1998-10-22 Revised:1998-12-15 Published:1999-10-15
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摘要：

测定了水、醋酸缓冲液和水杨苷醋酸 /醋酸钠缓冲液在水 /PC/庚烷-正丁醇反胶束体系中的饱和增溶量 ,利用UV、FT-IR和NMR等技术考察了增溶到水 /PC/庚烷-正丁醇反胶束体系中的 β-1 ,4葡萄糖苷酶及水杨苷的结构变化 ,并研究了 β-1 ,4葡萄糖苷酶在水 /PC/庚烷-正丁醇反胶束中的活性 .发现了 β-1 ,4葡萄糖苷酶在水 /PC/庚烷-正丁醇反胶束中遵循米氏反应 ,最大反应速度vmax为 1 .475mg•min-1,是其在水中的 11.8倍 ,米氏常数Km 为 0 .389mg•mL-1,是其在水中的 1 / 2 5 .β-1 ,4葡萄糖苷酶在水 /PC/庚烷-正丁醇反胶束中表现出的低底物浓度下的高催化速度 ,是微乳的界面发生的相互作用改变了 β-1 ,4葡萄糖苷酶的构象 ,增加了底物水杨苷的浓度 ,PC极性头部 -N+(CH3 ) 3 正离子作用水杨苷糖苷键的氧有利于糖苷键的断裂等因素造成的 .

关键词: 纤维素酶, 微乳, 增溶, 活性

Abstract:

 The maximum solubilization of water, acetic buffer and salicin in H 2O/PC/Heptan Butanol reverse micelles were measured. The change of structure of β-1,4-glucosidase and salicin in this system was investigated by UV ,FT IR and NMR. The activity and kinetics of β-1,4-glucosidase in this system were also studied. The results indicated that β-1,4-glucosidase follows the Michaelis-Menten kinetics in this reverse micelles, the v max is 1.475mg•min -1 ,11.8 times larger than that in aqueous, the Km is 0.389mg•mL -1 only 1/25 of that in aqueous. The reasons which cause this phenomenon are that the interaction at the interface of the reverse micelles changes the structure of β-1,4-glucosidase ,increases the concentration of salicin ,and that the -N +(CH 3) 3 in PC′ polar head attacks the oxygen of the glycoside bond of salicin .

Key words: β-1, 4-glucosidase, Phosphatidalcholine(PC), Reverse micelles, Solubilization, Activity