聚乙烯醇和海藻酸钠固定化柚苷酶的制备及其性质  

Preparation and Properties of PVA-Alginate Immobilized Naringinase

作　　者：肖安风[1,2,3,4] 游洪燕[1] 倪辉[1,2,3,4] 朱艳冰[1,2,3,4] 蔡慧农[1,2,3,4] 

机构地区：[1]集美大学生物工程学院福建,厦门361021 [2]福建省食品微生物与酶工程重点实验室,福建厦门361021 [3]福建省高校食品微生物与酶工程技术研究中心,福建厦门361021 [4]厦门市食品与生物工程技术研究中心,福建厦门361021

出　　处：《中国食品学报》2015年第3期15-23,共9页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家自然科学基金项目(31271914);福建省科技计划重点项目(2011N0101)

摘　　要：以聚乙烯醇、海藻酸钠为载体,固定化柚苷酶;以戊二醛为交联剂,考察固定化工艺条件对酶活的影响,研究固定化酶的部分酶学性质。用高效液相色谱法分析柚苷酶的α-L-鼠李糖苷酶和柚苷酶活力,结果表明:最佳载体组合为11%聚乙烯醇与0.5%海藻酸钠;当缓冲液p H 4、戊二醛含量1%、交联时间0.5 h、吸附时间3 h、加酶量141 U/m L、硼酸4%、Ca Cl21%时,固定化酶活力达到最大值。柚苷酶固定化后,α-L-鼠李糖苷酶和柚苷酶活力的最适温度提高5℃,最佳活性p H值提高1.0,p H稳定性基本不变,温度稳定性下降5℃。α-L-鼠李糖苷酶、柚苷酶的米氏常数(Km)固定化后都增大;α-L-鼠李糖苷酶的最大反应初速度(Vmax)经固定化后减小,柚苷酶的Vmax增大。将固定化柚苷酶重复使用7次后,它的α-L-鼠李糖苷酶和柚苷酶残余活力分别保持71%与80%。In this paper, naringinase was immobilized into matrix consisting of polyvinyl alcohol（PVA） and sodium alginate which were crosslinked with glutaraldehyde. The enzyme activity of naringinase and α-rhamnosidase was analysed by HPLC method. The optimal immobilized conditions were achieved with 11% PVA and 0.5% sodium alginate at p H 4,1% glutaraldehyde for 0.5 h, 141 U of enzyme load of per milliliter matrix, adsorption time of 3 h, 4% boric acid and1% Ca Cl2, respectively. In addition, the research for immobilized enzymology properties showed that the optimum temperature and p H of immobilized enzyme was higher than that of free form. Compared with free enzyme, p H stability revealed no change by immobilization, and instead, temperature stability of immobilized enzyme decreased 5 ℃. The Michaelis constant（Km） of naringinase and α-rhamnosidase were increased by immobilization, the maximum reaction velocity（Vmax） of naringinase and α-rhamnosidase were increased and decreased by immobilization, respectively.

关 键 词：聚乙烯醇 海藻酸钠 固定化 柚苷酶 α-鼠李糖苷酶 

分 类 号：TS201.25[轻工技术与工程—食品科学]