Suppr超能文献

用于嗜热聚羟基脂肪酸酯生产者的首个模型——嗜热解聚单胞菌的初始基因组编辑工具集。

An Initial Genome Editing Toolset for Caldimonas thermodepolymerans, the First Model of Thermophilic Polyhydroxyalkanoates Producer.

作者信息

Grybchuk-Ieremenko Anastasiia, Lipovská Kristýna, Kouřilová Xenie, Obruča Stanislav, Dvořák Pavel

机构信息

Department of Experimental Biology (Section of Microbiology), Faculty of Science, Masaryk University, Brno, Czech Republic.

Faculty of Chemistry, Institute of Food Science and Biotechnology, Brno University of Technology, Brno, Czech Republic.

出版信息

Microb Biotechnol. 2025 Feb;18(2):e70103. doi: 10.1111/1751-7915.70103.

Abstract

The limited number of well-characterised model bacteria cannot address all the challenges in a circular bioeconomy. Therefore, there is a growing demand for new production strains with enhanced resistance to extreme conditions, versatile metabolic capabilities and the ability to utilise cost-effective renewable resources while efficiently generating attractive biobased products. Particular thermophilic microorganisms fulfil these requirements. Non-virulent Gram-negative Caldimonas thermodepolymerans DSM15344 is one such attractive thermophile that efficiently converts a spectrum of plant biomass sugars into high quantities of polyhydroxyalkanoates (PHA)-a fully biodegradable substitutes for synthetic plastics. However, to enhance its biotechnological potential, the bacterium needs to be 'domesticated'. In this study, we established effective homologous recombination and transposon-based genome editing systems for C. thermodepolymerans. By optimising the electroporation protocol and refining counterselection methods, we achieved significant improvements in genetic manipulation and constructed the AI01 chassis strain with improved transformation efficiency and a ΔphaC mutant that will be used to study the importance of PHA synthesis in Caldimonas. The advances described herein highlight the need for tailored approaches when working with thermophilic bacteria and provide a springboard for further genetic and metabolic engineering of C. thermodepolymerans, which can be considered the first model of thermophilic PHA producer.

摘要

特征明确的模式细菌数量有限,无法应对循环生物经济中的所有挑战。因此,对新型生产菌株的需求日益增长,这些菌株需要增强对极端条件的抗性、具备多功能代谢能力,以及能够利用具有成本效益的可再生资源,同时高效生产有吸引力的生物基产品。某些嗜热微生物满足这些要求。无毒革兰氏阴性嗜热解聚卡尔迪单胞菌DSM15344就是这样一种有吸引力的嗜热菌,它能有效地将多种植物生物质糖转化为大量聚羟基脂肪酸酯(PHA)——一种可完全生物降解的合成塑料替代品。然而,为了增强其生物技术潜力,需要对这种细菌进行“驯化”。在本研究中,我们为嗜热解聚卡尔迪单胞菌建立了有效的同源重组和基于转座子的基因组编辑系统。通过优化电穿孔方案和改进反选择方法,我们在基因操作方面取得了显著进展,构建了转化效率提高的AI01底盘菌株和一个ΔphaC突变体,该突变体将用于研究PHA合成在卡尔迪单胞菌中的重要性。本文所述的进展凸显了处理嗜热细菌时采用定制方法的必要性,并为嗜热解聚卡尔迪单胞菌的进一步基因和代谢工程提供了一个跳板,嗜热解聚卡尔迪单胞菌可被视为嗜热PHA生产者的首个模型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/151f/11842462/ac3b16955454/MBT2-18-e70103-g004.jpg

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验