微型气泡色谱柱实现小型化缩放

IF 3.9 4区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Engineering in Life Sciences Pub Date : 2024-09-02 DOI:10.1002/elsc.202400051
Moritz Wild, Ralf Takors
{"title":"微型气泡色谱柱实现小型化缩放","authors":"Moritz Wild, Ralf Takors","doi":"10.1002/elsc.202400051","DOIUrl":null,"url":null,"abstract":"The successful scale‐up of biotechnological processes from laboratory to industrial scale is crucial for translating innovation to practice. Scale‐down simulators have emerged as indispensable tools in this endeavor, enabling the evaluation of potential hosts’ adaptability to the dynamic conditions encountered in large‐scale fermenters. By simulating these real‐world scenarios, scale‐down simulators facilitate more accurate estimations of host productivity, thereby improving the process of selecting optimal strains for industrial production. Conventional scale‐down systems for detailed intracellular analysis necessitate an elaborate setup comprising interconnected lab‐scale reactors such as stirred tank reactors (STRs) and plug‐flow reactors (PFRs), often proving time‐consuming and resource‐intensive. This work introduces a miniaturized bubble column reactor setup (60 mL working volume), enabling individual and parallel carbon‐limited chemostat fermentations, offering a more efficient and streamlined approach. The industrially relevant organism <jats:italic>Escherichia coli</jats:italic>, chosen as a model organism, is continuously grown and subjected to carbon starvation for 150 s, followed by a return to carbon excess for another 150 s. The cellular response is characterized by the accumulation of the alarmone guanosine pentaphosphate (ppGpp) accompanied by a significant reduction in energy charge, from 0.8 to 0.7, which is rapidly replenished upon reintroduction of carbon availability. Transcriptomic analysis reveals a two‐phase response pattern, with over 200 genes upregulated and downregulated. The initial phase is dominated by the CRP–cAMP‐ and ppGpp‐mediated response to carbon limitation, followed by a shift to stationary phase‐inducing gene expression under the control of stress sigma factors. The system's validity is confirmed through a thorough comparison with a conventional STR/PFR setup. The analysis reveals the potential of the system to effectively reproduce data gathered from conventional STR/PFR setups, showcasing its potential use as a scale‐down simulator integrated in the process of strain development.","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"436 1","pages":""},"PeriodicalIF":3.9000,"publicationDate":"2024-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Mini Bubble Columns for Miniaturizing Scale‐Down\",\"authors\":\"Moritz Wild, Ralf Takors\",\"doi\":\"10.1002/elsc.202400051\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"The successful scale‐up of biotechnological processes from laboratory to industrial scale is crucial for translating innovation to practice. Scale‐down simulators have emerged as indispensable tools in this endeavor, enabling the evaluation of potential hosts’ adaptability to the dynamic conditions encountered in large‐scale fermenters. By simulating these real‐world scenarios, scale‐down simulators facilitate more accurate estimations of host productivity, thereby improving the process of selecting optimal strains for industrial production. Conventional scale‐down systems for detailed intracellular analysis necessitate an elaborate setup comprising interconnected lab‐scale reactors such as stirred tank reactors (STRs) and plug‐flow reactors (PFRs), often proving time‐consuming and resource‐intensive. This work introduces a miniaturized bubble column reactor setup (60 mL working volume), enabling individual and parallel carbon‐limited chemostat fermentations, offering a more efficient and streamlined approach. The industrially relevant organism <jats:italic>Escherichia coli</jats:italic>, chosen as a model organism, is continuously grown and subjected to carbon starvation for 150 s, followed by a return to carbon excess for another 150 s. The cellular response is characterized by the accumulation of the alarmone guanosine pentaphosphate (ppGpp) accompanied by a significant reduction in energy charge, from 0.8 to 0.7, which is rapidly replenished upon reintroduction of carbon availability. Transcriptomic analysis reveals a two‐phase response pattern, with over 200 genes upregulated and downregulated. The initial phase is dominated by the CRP–cAMP‐ and ppGpp‐mediated response to carbon limitation, followed by a shift to stationary phase‐inducing gene expression under the control of stress sigma factors. The system's validity is confirmed through a thorough comparison with a conventional STR/PFR setup. The analysis reveals the potential of the system to effectively reproduce data gathered from conventional STR/PFR setups, showcasing its potential use as a scale‐down simulator integrated in the process of strain development.\",\"PeriodicalId\":11678,\"journal\":{\"name\":\"Engineering in Life Sciences\",\"volume\":\"436 1\",\"pages\":\"\"},\"PeriodicalIF\":3.9000,\"publicationDate\":\"2024-09-02\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Engineering in Life Sciences\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://doi.org/10.1002/elsc.202400051\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Engineering in Life Sciences","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1002/elsc.202400051","RegionNum":4,"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

摘要

成功地将生物技术过程从实验室放大到工业规模,对于将创新转化为实践至关重要。缩小规模模拟器已成为这项工作中不可或缺的工具,可评估潜在宿主对大型发酵罐中动态条件的适应性。通过模拟这些真实世界的场景,缩比模拟器有助于更准确地估计宿主的生产率,从而改进为工业生产选择最佳菌株的过程。用于详细细胞内分析的传统缩比系统需要复杂的设置,包括相互连接的实验室级反应器,如搅拌罐反应器(STR)和塞流反应器(PFR),往往耗费大量时间和资源。这项研究引入了一种小型化的气泡柱反应器装置(工作容积为 60 毫升),可进行单独和并行的碳限制恒温发酵,提供了一种更高效、更简化的方法。细胞反应的特征是报警酮五磷酸鸟苷(ppGpp)的积累,同时伴随着能量电荷的显著降低,从 0.8 降至 0.7。转录组分析显示了两阶段的反应模式,有 200 多个基因上调和下调。初始阶段以 CRP-cAMP 和 ppGpp 介导的碳限制反应为主,随后在应激 sigma 因子的控制下转入静止阶段,诱导基因表达。通过与传统的 STR/PFR 设置进行全面比较,证实了该系统的有效性。分析结果表明,该系统具有有效再现从传统 STR/PFR 设置中收集的数据的潜力,展示了其作为集成在菌株开发过程中的缩放模拟器的潜在用途。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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
Mini Bubble Columns for Miniaturizing Scale‐Down
The successful scale‐up of biotechnological processes from laboratory to industrial scale is crucial for translating innovation to practice. Scale‐down simulators have emerged as indispensable tools in this endeavor, enabling the evaluation of potential hosts’ adaptability to the dynamic conditions encountered in large‐scale fermenters. By simulating these real‐world scenarios, scale‐down simulators facilitate more accurate estimations of host productivity, thereby improving the process of selecting optimal strains for industrial production. Conventional scale‐down systems for detailed intracellular analysis necessitate an elaborate setup comprising interconnected lab‐scale reactors such as stirred tank reactors (STRs) and plug‐flow reactors (PFRs), often proving time‐consuming and resource‐intensive. This work introduces a miniaturized bubble column reactor setup (60 mL working volume), enabling individual and parallel carbon‐limited chemostat fermentations, offering a more efficient and streamlined approach. The industrially relevant organism Escherichia coli, chosen as a model organism, is continuously grown and subjected to carbon starvation for 150 s, followed by a return to carbon excess for another 150 s. The cellular response is characterized by the accumulation of the alarmone guanosine pentaphosphate (ppGpp) accompanied by a significant reduction in energy charge, from 0.8 to 0.7, which is rapidly replenished upon reintroduction of carbon availability. Transcriptomic analysis reveals a two‐phase response pattern, with over 200 genes upregulated and downregulated. The initial phase is dominated by the CRP–cAMP‐ and ppGpp‐mediated response to carbon limitation, followed by a shift to stationary phase‐inducing gene expression under the control of stress sigma factors. The system's validity is confirmed through a thorough comparison with a conventional STR/PFR setup. The analysis reveals the potential of the system to effectively reproduce data gathered from conventional STR/PFR setups, showcasing its potential use as a scale‐down simulator integrated in the process of strain development.
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
Engineering in Life Sciences
Engineering in Life Sciences 工程技术-生物工程与应用微生物
CiteScore
6.40
自引率
3.70%
发文量
81
审稿时长
3 months
期刊介绍: Engineering in Life Sciences (ELS) focuses on engineering principles and innovations in life sciences and biotechnology. Life sciences and biotechnology covered in ELS encompass the use of biomolecules (e.g. proteins/enzymes), cells (microbial, plant and mammalian origins) and biomaterials for biosynthesis, biotransformation, cell-based treatment and bio-based solutions in industrial and pharmaceutical biotechnologies as well as in biomedicine. ELS especially aims to promote interdisciplinary collaborations among biologists, biotechnologists and engineers for quantitative understanding and holistic engineering (design-built-test) of biological parts and processes in the different application areas.
期刊最新文献
Issue Information Cover Picture: Engineering in Life Sciences 11'24 Mechanical Microvibration Device Enhancing Immunohistochemistry Efficiency Issue Information Cover Picture: Engineering in Life Sciences 10'24
×
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