{"title":"Structures and properties of α-amylase and glucoamylase immobilized by ZIF-8 via one-pot preparation.","authors":"Yuxin Liu, Qinghua Pan, Zesheng Liang, Jingqiao Li, Rulong Wu","doi":"10.1016/j.enzmictec.2024.110579","DOIUrl":null,"url":null,"abstract":"<p><p>The immobilization of α-amylase and glucoamylase using a metal-organic framework (enzyme@ZIF-8) was prepared in situ through a one-pot method. The morphology, crystal structure, and molecular characteristics of the free enzyme and enzyme@ZIF-8 were characterized. The enzyme@ZIF-8 exhibited the rhombic dodecahedron morphology, with a decrease in particle size. Successful immobilization of α-amylase and glucoamylase within ZIF-8 was confirmed, with 30-40 % loading rate. The immobilization process did not significantly alter the crystal structure of ZIF-8. The changes in secondary structure of enzyme after immobilization resulted in modification of catalytic activity of enzyme. The melting enthalpy of enzyme @ZIF-8 increased with the increase of enzyme content. The melting peak temperature of the enzyme immobilized by ZIF-8 increased. The activity of free and immobilized enzymes was influenced by the different time, pH and temperature. At pH 5-8 and temperature 60-80 °C, the activity of the immobilized enzyme was significantly greater than that of the free enzyme. The repeatability of enzyme@ZIF-8 was 61.52 % after three cycles. The kinetic parameters of Michaelis-Menten model for enzymatic reaction were determined by fitting the initial rate of reactions and initial substrate concentration data. The Michaelis-Menten constant (K<sub>M</sub>) values of immobilized enzyme were lower than that of free enzyme, indicating the greater affinity between the enzyme and the substrate.</p>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"184 ","pages":"110579"},"PeriodicalIF":3.4000,"publicationDate":"2024-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Enzyme and Microbial Technology","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1016/j.enzmictec.2024.110579","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
The immobilization of α-amylase and glucoamylase using a metal-organic framework (enzyme@ZIF-8) was prepared in situ through a one-pot method. The morphology, crystal structure, and molecular characteristics of the free enzyme and enzyme@ZIF-8 were characterized. The enzyme@ZIF-8 exhibited the rhombic dodecahedron morphology, with a decrease in particle size. Successful immobilization of α-amylase and glucoamylase within ZIF-8 was confirmed, with 30-40 % loading rate. The immobilization process did not significantly alter the crystal structure of ZIF-8. The changes in secondary structure of enzyme after immobilization resulted in modification of catalytic activity of enzyme. The melting enthalpy of enzyme @ZIF-8 increased with the increase of enzyme content. The melting peak temperature of the enzyme immobilized by ZIF-8 increased. The activity of free and immobilized enzymes was influenced by the different time, pH and temperature. At pH 5-8 and temperature 60-80 °C, the activity of the immobilized enzyme was significantly greater than that of the free enzyme. The repeatability of enzyme@ZIF-8 was 61.52 % after three cycles. The kinetic parameters of Michaelis-Menten model for enzymatic reaction were determined by fitting the initial rate of reactions and initial substrate concentration data. The Michaelis-Menten constant (KM) values of immobilized enzyme were lower than that of free enzyme, indicating the greater affinity between the enzyme and the substrate.
期刊介绍：
Enzyme and Microbial Technology is an international, peer-reviewed journal publishing original research and reviews, of biotechnological significance and novelty, on basic and applied aspects of the science and technology of processes involving the use of enzymes, micro-organisms, animal cells and plant cells.
We especially encourage submissions on:
Biocatalysis and the use of Directed Evolution in Synthetic Biology and Biotechnology
Biotechnological Production of New Bioactive Molecules, Biomaterials, Biopharmaceuticals, and Biofuels
New Imaging Techniques and Biosensors, especially as applicable to Healthcare and Systems Biology
New Biotechnological Approaches in Genomics, Proteomics and Metabolomics
Metabolic Engineering, Biomolecular Engineering and Nanobiotechnology
Manuscripts which report isolation, purification, immobilization or utilization of organisms or enzymes which are already well-described in the literature are not suitable for publication in EMT, unless their primary purpose is to report significant new findings or approaches which are of broad biotechnological importance. Similarly, manuscripts which report optimization studies on well-established processes are inappropriate. EMT does not accept papers dealing with mathematical modeling unless they report significant, new experimental data.
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