通过合理工程改善菊粉酶的活性,以高效生物合成低分子量菊粉。

IF 2.3 3区 生物学 Q3 MICROBIOLOGY
Dawei Ni, Zhaolin Huang, Shuqi Zhang, Yang Yang, Xiaoyong Liu, Wei Xu, Wenli Zhang, Wanmeng Mu
{"title":"通过合理工程改善菊粉酶的活性,以高效生物合成低分子量菊粉。","authors":"Dawei Ni,&nbsp;Zhaolin Huang,&nbsp;Shuqi Zhang,&nbsp;Yang Yang,&nbsp;Xiaoyong Liu,&nbsp;Wei Xu,&nbsp;Wenli Zhang,&nbsp;Wanmeng Mu","doi":"10.1007/s00203-024-04153-7","DOIUrl":null,"url":null,"abstract":"<div><p>Inulin, a widely recognized prebiotic, has diverse applications across various industrial sectors. Although inulin is primarily produced through plant extraction, there is growing interest in enzymatic synthesis as an alternative. The enzymatic production of inulin from sucrose, which yields polymers with degrees of polymerization similar to those of plant-derived inulin, shows potential as a viable replacement for traditional extraction methods. In this study, an inulosucrase from <i>Neobacillus bataviensis</i> was identified, demonstrating a non-processive mechanism specifically tailored for synthesizing inulin with polymerization degrees ranging from 3 to approximately 40. The enzyme exhibited optimal activity at pH 6.5 and 55 °C, efficiently producing inulin with a yield of 50.6%. Ca<sup>2+</sup> can improve the activity and thermostability of this enzyme. To enhance catalytic total activity, site-directed and truncated mutagenesis techniques were applied, resulting in the identification of a mutant, T149S, displaying a significant 57% increase in catalytic total activity. Molecular dynamics simulations unveiled that the heightened flexibility observed in three surface regions positively influenced enzymatic activity. This study not only contributes to the theoretical foundation for inulosucrase engineering but also presents a potential avenue for the production of inulin.</p></div>","PeriodicalId":8279,"journal":{"name":"Archives of Microbiology","volume":"206 11","pages":""},"PeriodicalIF":2.3000,"publicationDate":"2024-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Improving the activity of an inulosucrase by rational engineering for the efficient biosynthesis of low-molecular-weight inulin\",\"authors\":\"Dawei Ni,&nbsp;Zhaolin Huang,&nbsp;Shuqi Zhang,&nbsp;Yang Yang,&nbsp;Xiaoyong Liu,&nbsp;Wei Xu,&nbsp;Wenli Zhang,&nbsp;Wanmeng Mu\",\"doi\":\"10.1007/s00203-024-04153-7\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Inulin, a widely recognized prebiotic, has diverse applications across various industrial sectors. Although inulin is primarily produced through plant extraction, there is growing interest in enzymatic synthesis as an alternative. The enzymatic production of inulin from sucrose, which yields polymers with degrees of polymerization similar to those of plant-derived inulin, shows potential as a viable replacement for traditional extraction methods. In this study, an inulosucrase from <i>Neobacillus bataviensis</i> was identified, demonstrating a non-processive mechanism specifically tailored for synthesizing inulin with polymerization degrees ranging from 3 to approximately 40. The enzyme exhibited optimal activity at pH 6.5 and 55 °C, efficiently producing inulin with a yield of 50.6%. Ca<sup>2+</sup> can improve the activity and thermostability of this enzyme. To enhance catalytic total activity, site-directed and truncated mutagenesis techniques were applied, resulting in the identification of a mutant, T149S, displaying a significant 57% increase in catalytic total activity. Molecular dynamics simulations unveiled that the heightened flexibility observed in three surface regions positively influenced enzymatic activity. This study not only contributes to the theoretical foundation for inulosucrase engineering but also presents a potential avenue for the production of inulin.</p></div>\",\"PeriodicalId\":8279,\"journal\":{\"name\":\"Archives of Microbiology\",\"volume\":\"206 11\",\"pages\":\"\"},\"PeriodicalIF\":2.3000,\"publicationDate\":\"2024-10-03\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Archives of Microbiology\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://link.springer.com/article/10.1007/s00203-024-04153-7\",\"RegionNum\":3,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Archives of Microbiology","FirstCategoryId":"99","ListUrlMain":"https://link.springer.com/article/10.1007/s00203-024-04153-7","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0

摘要

菊粉是一种被广泛认可的益生元,在各个工业领域都有不同的应用。虽然菊粉主要是通过植物提取生产的,但人们对酶法合成这种替代方法的兴趣日益浓厚。从蔗糖中用酶法生产菊粉,可获得聚合度与植物菊粉相似的聚合物,显示出替代传统提取方法的潜力。本研究从巴达维新杆菌(Neobacillus bataviensis)中发现了一种菊粉蔗糖酶,它展示了一种非加工机制,专门用于合成聚合度从 3 到约 40 的菊粉。该酶在 pH 值为 6.5 和温度为 55 ℃ 时表现出最佳活性,能高效生产菊粉,产量为 50.6%。Ca2+ 可以提高这种酶的活性和耐热性。为了提高催化总活性,研究人员采用了定点突变和截断突变技术,最终确定了一个突变体 T149S,其催化总活性显著提高了 57%。分子动力学模拟揭示了在三个表面区域观察到的灵活性增强对酶活性的积极影响。这项研究不仅为菊粉酶工程学奠定了理论基础,还为菊粉的生产提供了一条潜在的途径。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Improving the activity of an inulosucrase by rational engineering for the efficient biosynthesis of low-molecular-weight inulin

Inulin, a widely recognized prebiotic, has diverse applications across various industrial sectors. Although inulin is primarily produced through plant extraction, there is growing interest in enzymatic synthesis as an alternative. The enzymatic production of inulin from sucrose, which yields polymers with degrees of polymerization similar to those of plant-derived inulin, shows potential as a viable replacement for traditional extraction methods. In this study, an inulosucrase from Neobacillus bataviensis was identified, demonstrating a non-processive mechanism specifically tailored for synthesizing inulin with polymerization degrees ranging from 3 to approximately 40. The enzyme exhibited optimal activity at pH 6.5 and 55 °C, efficiently producing inulin with a yield of 50.6%. Ca2+ can improve the activity and thermostability of this enzyme. To enhance catalytic total activity, site-directed and truncated mutagenesis techniques were applied, resulting in the identification of a mutant, T149S, displaying a significant 57% increase in catalytic total activity. Molecular dynamics simulations unveiled that the heightened flexibility observed in three surface regions positively influenced enzymatic activity. This study not only contributes to the theoretical foundation for inulosucrase engineering but also presents a potential avenue for the production of inulin.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
Archives of Microbiology
Archives of Microbiology 生物-微生物学
CiteScore
4.90
自引率
3.60%
发文量
601
审稿时长
3 months
期刊介绍: Research papers must make a significant and original contribution to microbiology and be of interest to a broad readership. The results of any experimental approach that meets these objectives are welcome, particularly biochemical, molecular genetic, physiological, and/or physical investigations into microbial cells and their interactions with their environments, including their eukaryotic hosts. Mini-reviews in areas of special topical interest and papers on medical microbiology, ecology and systematics, including description of novel taxa, are also published. Theoretical papers and those that report on the analysis or ''mining'' of data are acceptable in principle if new information, interpretations, or hypotheses emerge.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信