食品与发酵工业 >

2022 , Vol. 48 >Issue 19: 9 - 15

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.031520

研究报告

硅纳米粒子吸附交联固定化蔗糖异构酶的制备及其酶学性能研究

  • 陈宁 ,
  • 常保根 ,
  • 施念 ,
  • 路福平 ,
  • 刘夫锋
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  • (工业发酵微生物教育部重点实验室,天津市工业微生物重点实验室,工业酶国家工程实验室,天津科技大学 生物工程学院,天津,300457)
第一作者：博士,助理研究员(路福平教授和刘夫锋教授为共同通信作者,E-mail:lfp@tust.edu.cn;fufengliu@tust.edu.cn)

收稿日期: 2022-03-11

  修回日期: 2021-03-21

  网络出版日期: 2022-11-01

基金资助

国家重点研发专项(2021YFC2102700);国家自然科学基金项目(32272269)

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Preparation and characterization of adsorption-crosslinking immobilized sucrose isomerase

  • CHEN Ning ,
  • CHANG Baogen ,
  • SHI Nian ,
  • LU Fuping ,
  • LIU Fufeng
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  • (Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education;Tianjin Key Laboratory of Industrial Microbiology;National Engineering Laboratory for Industrial Enzymes;College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300457, China)

Received date: 2022-03-11

  Revised date: 2021-03-21

  Online published: 2022-11-01

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摘要

利用蔗糖异构酶(sucrose isomerase,SIase)异构催化蔗糖是目前生产异麦芽酮糖最常用的方法。以硅纳米粒子(silica nanoparticle,SNP)为载体、戊二醛(glutaraldehyde,GA)为交联剂,采用吸附法和吸附-交联法对蔗糖异构酶进行固定化,分别得到S-CLEAs和S-CLEAs-GA两种固定化酶。实验结果表明,固定化的最优条件是:硅纳米粒子6.25 mg/mL,酶加量8.6 U/mL,吸附时间为5 h,戊二醛的质量分数为0.08%,交联时间为2 h,此时S-CLEAs和S-CLEAs-GA的酶活力回收率分别为51.2%和44.9%。与游离酶相比,2种固定化酶在较宽的pH和温度范围内始终保持较高的生物催化活性。另外,S-CLEAs和S-CLEAs-GA重复使用15次后,酶活力回收率分别为55.1%和77.9%。表明SIase经此方法固定化后,固定化酶具有优异的热稳定性、pH耐受性和操作稳定性,其中S-CLEAs-GA的表现更好,有良好的工业应用前景。

关键词： 蔗糖异构酶; 固定化; 吸附; 交联; 硅纳米粒子

本文引用格式

陈宁 , 常保根 , 施念 , 路福平 , 刘夫锋 . 硅纳米粒子吸附交联固定化蔗糖异构酶的制备及其酶学性能研究[J]. 食品与发酵工业, 2022 , 48(19) : 9 -15 . DOI: 10.13995/j.cnki.11-1802/ts.031520

Abstract

Isomaltulose is a natural isomer of sucrose. It is widely used as a functional sweetener with promising properties, including slower digestion, prolonged energy release, lower insulin reaction, and less cariogenicity. The isomaltulose production by sucrose isomerase (SIase) isomerization catalyzing sucrose to produce isomaltulose is the most common method at present. SIase was immobilized on silica nanoparticle (SNP) cross-linked with glutaraldehyde (GA) by adsorption and adsorption-crosslinking method, and two immobilized enzyme, S-CLEAs and S-CLEAs-GA were obtained. The enzyme activity recovery of S-CLEAs reached up to 51.2% under the optimal condition of immobilization achieved with 6.25 mg/mL of SNP, 8.6 U/mL of SIase and 5 h of adsorption time. Then adding 0.08% GA and cross-linking 2 h, While the maximum activity recovery of S-CLEAs-GA was 44.9% when the GA concentration was 0.08% and cross-linking time was 2 h. Compared with free SIase, immobilized enzymes were maintained a relatively high enzyme activity in wide temperature and pH ranges. In addition, after 15 repeated uses, S-CLEAs-GA and S-CLEAs still had 55.1% and 77.9% of initial enzyme activity respectively. The above research results showed that immobilized SIase had excellent thermal, pH and operational stabilities, especially S-CLEAs-GA, thus had a good prospect of industrial application。

Key words： sucrose isomerase; immobilization; adsorption; crosslinking; silica nanoparticle

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