Sampling-free investigation of microbial carbon source preferences on renewable feedstocks via online monitoring of oxygen transfer rate.

IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Luca Antonia Grebe, Paul Richter, Torben Altenkirch, Marcel Mann, Markus Jan Müller, Jochen Büchs, Jørgen Barsett Magnus
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引用次数: 0

Abstract

The transition towards sustainable bioprocesses requires renewable feedstocks to reduce dependency on finite resources. While plant-based feedstocks offer significant potential, their complex composition poses new challenges. The microorganisms often exhibit polyauxic growth when presented with multiple carbon sources simultaneously, consuming them in a distinct order according to their carbon source preferences. The traditional investigation of polyauxic growth involves laborious sampling and offline analysis, hindering high-throughput screenings. This study introduces an efficient method for identifying carbon source consumption and their order of metabolization by various microorganisms using the respiration activity monitoring system (RAMOS) in shake flasks. As aerobic carbon metabolization and oxygen consumption are strictly correlated, the characteristic phases of polyauxic growth are visible in the oxygen transfer rate (OTR) and can be assigned to the respective carbon sources. An extended 16-flask RAMOS enables real-time monitoring of microbial respiration on up to seven carbon sources and one reference cultivation simultaneously, thus providing crucial insights into their metabolization without extensive sampling and offline analysis. The method's accuracy was validated against traditional high-performance liquid chromatography (HPLC). Its applicability to both fast-growing Escherichia coli (investigated carbon sources: glucose, arabinose, sorbitol, xylose, and glycerol) and slow-growing Ustilago trichophora (glucose, glycerol, xylose, sorbitol, rhamnose, galacturonic acid, and lactic acid) was demonstrated. Additionally, it was successfully applied to the plant-based second-generation feedstock corn leaf hydrolysate, revealing the bioavailability of the included carbon sources (glucose, sucrose, arabinose, xylose, and galactose) and their order of metabolization by Ustilago maydis.

通过在线监测氧转移率,对可再生原料中微生物碳源偏好进行免采样调查。
向可持续生物工艺过渡需要可再生原料,以减少对有限资源的依赖。虽然植物性原料具有巨大的潜力,但其复杂的成分也带来了新的挑战。当同时面对多种碳源时,微生物通常会表现出多氧生长,并根据其对碳源的偏好以不同的顺序消耗这些碳源。传统的多氧生长研究涉及费力的取样和离线分析,阻碍了高通量筛选。本研究介绍了一种有效的方法,利用摇瓶中的呼吸活动监测系统(RAMOS)识别各种微生物的碳源消耗及其代谢顺序。由于有氧碳代谢和氧消耗密切相关,多氧生长的特征阶段在氧转移率(OTR)中清晰可见,并可分配给相应的碳源。扩展的 16 瓶 RAMOS 可同时实时监测多达 7 种碳源和 1 种参考培养物的微生物呼吸,因此无需大量取样和离线分析即可深入了解其代谢情况。与传统的高效液相色谱法(HPLC)相比,该方法的准确性得到了验证。该方法适用于快速生长的大肠杆菌(研究碳源:葡萄糖、阿拉伯糖、山梨糖醇、木糖和甘油)和缓慢生长的三叶虫(葡萄糖、甘油、木糖、山梨糖醇、鼠李糖、半乳糖醛酸和乳酸)。此外,该方法还成功地应用于以植物为基础的第二代原料玉米叶水解物,揭示了所含碳源(葡萄糖、蔗糖、阿拉伯糖、木糖和半乳糖)的生物利用率以及麦角菌代谢这些碳源的顺序。
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来源期刊
Bioprocess and Biosystems Engineering
Bioprocess and Biosystems Engineering 工程技术-工程:化工
CiteScore
7.90
自引率
2.60%
发文量
147
审稿时长
2.6 months
期刊介绍: Bioprocess and Biosystems Engineering provides an international peer-reviewed forum to facilitate the discussion between engineering and biological science to find efficient solutions in the development and improvement of bioprocesses. The aim of the journal is to focus more attention on the multidisciplinary approaches for integrative bioprocess design. Of special interest are the rational manipulation of biosystems through metabolic engineering techniques to provide new biocatalysts as well as the model based design of bioprocesses (up-stream processing, bioreactor operation and downstream processing) that will lead to new and sustainable production processes. Contributions are targeted at new approaches for rational and evolutive design of cellular systems by taking into account the environment and constraints of technical production processes, integration of recombinant technology and process design, as well as new hybrid intersections such as bioinformatics and process systems engineering. Manuscripts concerning the design, simulation, experimental validation, control, and economic as well as ecological evaluation of novel processes using biosystems or parts thereof (e.g., enzymes, microorganisms, mammalian cells, plant cells, or tissue), their related products, or technical devices are also encouraged. The Editors will consider papers for publication based on novelty, their impact on biotechnological production and their contribution to the advancement of bioprocess and biosystems engineering science. Submission of papers dealing with routine aspects of bioprocess engineering (e.g., routine application of established methodologies, and description of established equipment) are discouraged.
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