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用于天然生产者乳酸乳球菌和清酒乳杆菌中细菌素产生特性分析的自动化工作流程。

Automated workflow for characterization of bacteriocin production in natural producers Lactococcus lactis and Latilactobacillus sakei.

作者信息

Steier Valentin, Prigolovkin Lisa, Reiter Alexander, Neddermann Tobias, Wiechert Wolfgang, Reich Sebastian J, Riedel Christian U, Oldiges Marco

机构信息

Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, Jülich, Germany.

Institute of Biotechnology, RWTH Aachen University, Aachen, Germany.

出版信息

Microb Cell Fact. 2024 Mar 3;23(1):74. doi: 10.1186/s12934-024-02349-6.

Abstract

BACKGROUND

Lactic acid bacteria are commonly used as protective starter cultures in food products. Among their beneficial effects is the production of ribosomally synthesized peptides termed bacteriocins that kill or inhibit food-spoiling bacteria and pathogens, e.g., members of the Listeria species. As new bacteriocins and producer strains are being discovered rapidly, modern automated methods for strain evaluation and bioprocess development are required to accelerate screening and development processes.

RESULTS

In this study, we developed an automated workflow for screening and bioprocess optimization for bacteriocin producing lactic acid bacteria, consisting of microcultivation, sample processing and automated antimicrobial activity assay. We implemented sample processing workflows to minimize bacteriocin adsorption to producer cells via addition of Tween 80 and divalent cations to the cultivation media as well as acidification of culture broth prior to cell separation. Moreover, we demonstrated the applicability of the automated workflow to analyze influence of media components such as MES buffer or yeast extract for bacteriocin producers Lactococcus lactis B1629 and Latilactobacillus sakei A1608.

CONCLUSIONS

Our automated workflow provides advanced possibilities to accelerate screening and bioprocess optimization for natural bacteriocin producers. Based on its modular concept, adaptations for other strains, bacteriocin products and applications are easily carried out and a unique tool to support bacteriocin research and bioprocess development is provided.

摘要

背景

乳酸菌常用于食品中的保护性发酵剂培养物。它们的有益作用之一是产生核糖体合成的肽,即细菌素,可杀死或抑制导致食品变质的细菌和病原体,例如李斯特菌属的成员。随着新的细菌素和产生菌株迅速被发现,需要现代自动化方法来进行菌株评估和生物工艺开发,以加速筛选和开发过程。

结果

在本研究中,我们开发了一种用于筛选和优化细菌素产生乳酸菌生物工艺的自动化工作流程,包括微量培养、样品处理和自动化抗菌活性测定。我们实施了样品处理工作流程,通过向培养基中添加吐温80和二价阳离子以及在细胞分离前对培养液进行酸化,以尽量减少细菌素对产生菌细胞的吸附。此外,我们证明了该自动化工作流程适用于分析培养基成分(如MES缓冲液或酵母提取物)对细菌素产生菌乳酸乳球菌B1629和清酒乳杆菌A1608的影响。

结论

我们的自动化工作流程为加速天然细菌素产生菌的筛选和生物工艺优化提供了先进的可能性。基于其模块化概念,可轻松对其他菌株、细菌素产品和应用进行调整,并提供了一个支持细菌素研究和生物工艺开发的独特工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8f12/10910668/5f17f1c49700/12934_2024_2349_Fig1_HTML.jpg

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