Improving Fructooligosaccharide Production via sacC Gene Deletion in Zymomonas mobilis: A Novel Approach for Enhanced Prebiotic Production

IF 5.3 2区 农林科学 Q1 FOOD SCIENCE & TECHNOLOGY Food and Bioprocess Technology Pub Date : 2024-07-12 DOI:10.1007/s11947-024-03508-8
Adelaide Braga, Ana Benedita Maia, Daniela Gomes, Joana L. Rodrigues, João Rainha, Lígia R. Rodrigues
{"title":"Improving Fructooligosaccharide Production via sacC Gene Deletion in Zymomonas mobilis: A Novel Approach for Enhanced Prebiotic Production","authors":"Adelaide Braga, Ana Benedita Maia, Daniela Gomes, Joana L. Rodrigues, João Rainha, Lígia R. Rodrigues","doi":"10.1007/s11947-024-03508-8","DOIUrl":null,"url":null,"abstract":"<p>Fructooligosaccharides (FOS) are promising prebiotics in the relevant and increasing market of functional food. Industrially, these compounds are produced from sucrose by the action of fructosyltransferase or b-fructofuranosidase enzymes. However, this process often yields low conversion rates and results in impure mixtures due to the release of high levels of glucose. <i>Zymomonas mobilis</i> is a well-known ethanol-producing bacterium with native levansucrase enzymes able to convert sucrose into FOS. This study aimed to use synthetic biology tools to eliminate invertase (<i>sacC</i>) activity in <i>Z. mobilis</i>, reducing substrate competition and maximizing FOS production. Additionally, we explored the potential use of agro-industrial by-products, such as sugarcane molasses (M) and corn step liquor (CSL), as nutrients for FOS production using <i>Z. mobilis</i> in an in vivo bioprocess strategy. Invertase deletion from the <i>Z. mobilis</i> ZM4 genome was accomplished by homologous recombination of an engineered suicide plasmid. Using <i>Z. mobilis sacC</i><sup><i>−</i></sup>, we observed a 70% reduction in monosaccharide production and a 9.0-fold increase in levan formation compared to the wild-type strain. Implementation of a fed-batch approach with CSL and molasses (CSLM) medium at flask-scale allowed to produce 41.9 g L<sup>−1</sup> of FOS (0.25 g<sub>FOS</sub> g<sub>sucrose</sub><sup>−1</sup>). To our knowledge, this work describes for the first time the production of FOS from agro-waste residues using a genetically modified <i>Z. mobilis</i> strain in a one-step fermentation. Through this innovative approach, we aim to contribute to the advancement of biotechnological strategies for prebiotic production, offering insights into genetic engineering techniques for improving the efficiency and sustainability of FOS synthesis in <i>Z. mobilis</i>.\n</p>","PeriodicalId":562,"journal":{"name":"Food and Bioprocess Technology","volume":"80 1","pages":""},"PeriodicalIF":5.3000,"publicationDate":"2024-07-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Food and Bioprocess Technology","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1007/s11947-024-03508-8","RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"FOOD SCIENCE & TECHNOLOGY","Score":null,"Total":0}
引用次数: 0

Abstract

Fructooligosaccharides (FOS) are promising prebiotics in the relevant and increasing market of functional food. Industrially, these compounds are produced from sucrose by the action of fructosyltransferase or b-fructofuranosidase enzymes. However, this process often yields low conversion rates and results in impure mixtures due to the release of high levels of glucose. Zymomonas mobilis is a well-known ethanol-producing bacterium with native levansucrase enzymes able to convert sucrose into FOS. This study aimed to use synthetic biology tools to eliminate invertase (sacC) activity in Z. mobilis, reducing substrate competition and maximizing FOS production. Additionally, we explored the potential use of agro-industrial by-products, such as sugarcane molasses (M) and corn step liquor (CSL), as nutrients for FOS production using Z. mobilis in an in vivo bioprocess strategy. Invertase deletion from the Z. mobilis ZM4 genome was accomplished by homologous recombination of an engineered suicide plasmid. Using Z. mobilis sacC, we observed a 70% reduction in monosaccharide production and a 9.0-fold increase in levan formation compared to the wild-type strain. Implementation of a fed-batch approach with CSL and molasses (CSLM) medium at flask-scale allowed to produce 41.9 g L−1 of FOS (0.25 gFOS gsucrose−1). To our knowledge, this work describes for the first time the production of FOS from agro-waste residues using a genetically modified Z. mobilis strain in a one-step fermentation. Through this innovative approach, we aim to contribute to the advancement of biotechnological strategies for prebiotic production, offering insights into genetic engineering techniques for improving the efficiency and sustainability of FOS synthesis in Z. mobilis.

Abstract Image

查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
通过删除 Zymomonas mobilis 中的 sacC 基因提高果寡糖产量:提高益生菌生产的新方法
果寡糖(FOS)是一种前景广阔的益生元,在相关的功能性食品市场中占有越来越大的份额。在工业上,这些化合物是通过果糖基转移酶或 b-呋喃果糖酶的作用从蔗糖中生产出来的。然而,由于释放出大量葡萄糖,这一过程的转化率往往较低,并导致混合物不纯。Zymomonas mobilis 是一种著名的乙醇生产细菌,其原生左旋蔗糖酶能够将蔗糖转化为 FOS。本研究旨在利用合成生物学工具消除莫比莱氏单胞菌(Z. mobilis)中的转化酶(sacC)活性,从而减少底物竞争,最大限度地提高 FOS 产量。此外,我们还探索了利用甘蔗糖蜜(M)和玉米阶梯液(CSL)等农用工业副产品作为营养物质的潜力,并在体内生物加工策略中使用莫比莱氏菌生产 FOS。通过工程自杀质粒的同源重组,实现了莫比莱氏菌 ZM4 基因组中反转酶的缺失。通过使用 Z. mobilis sacC-,我们观察到与野生型菌株相比,单糖产量减少了 70%,levan 的形成增加了 9.0 倍。在烧瓶规模下使用 CSL 和糖蜜(CSLM)培养基的喂养批次方法可生产 41.9 g L-1 的 FOS(0.25 gFOS gsucrose-1)。据我们所知,这项工作首次描述了利用转基因 Z. mobilis 菌株一步发酵法从农业废弃物残渣中生产 FOS 的过程。通过这一创新方法,我们旨在为益生元生产的生物技术战略的进步做出贡献,为提高莫比莱氏菌合成 FOS 的效率和可持续性的基因工程技术提供见解。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 去求助
来源期刊
Food and Bioprocess Technology
Food and Bioprocess Technology 农林科学-食品科技
CiteScore
9.50
自引率
19.60%
发文量
200
审稿时长
2.8 months
期刊介绍: Food and Bioprocess Technology provides an effective and timely platform for cutting-edge high quality original papers in the engineering and science of all types of food processing technologies, from the original food supply source to the consumer’s dinner table. It aims to be a leading international journal for the multidisciplinary agri-food research community. The journal focuses especially on experimental or theoretical research findings that have the potential for helping the agri-food industry to improve process efficiency, enhance product quality and, extend shelf-life of fresh and processed agri-food products. The editors present critical reviews on new perspectives to established processes, innovative and emerging technologies, and trends and future research in food and bioproducts processing. The journal also publishes short communications for rapidly disseminating preliminary results, letters to the Editor on recent developments and controversy, and book reviews.
期刊最新文献
Production and Preliminary Characterization of Microbial Cellulose Generated in Fermented Moringa and Coated with Flaxseed Mucilage Pomegranate Peel Flour as a Co-encapsulant Improves the Survival of Lactic Acid Bacteria to Thermal Treatment and Simulated Gastrointestinal Conditions A Novel Approach for Solubility and Bioavailability Enhancement of Canthaxanthin Obtained from Dietzia natronolimnaea HS-1 by Canthaxanthin-V-Amylose Complex Storage Stability and Anthocyanin Degradation Kinetics of Alginate/Carboxymethyl Cellulose–based Emulsion Fortified with Red Onion Peel Extracts and its Application in the Preservation of Fresh Strawberries Investigating the Impact of Hydrophobic Deep Eutectic Oil-in-Water Nanoemulsion on Cell Membrane Degradation and Inhibition of C. gloeosporioides in Postharvest Technology
×
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
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 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 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1