Engineering Bacillus subtilis J46 for efficient utilization of galactose through adaptive laboratory evolution

AMB Express, Journal Year: 2024, Volume and Issue: 14(1)

Published: Jan. 29, 2024

Efficient utilization of galactose by microorganisms can lead to the production valuable bio-products and improved metabolic processes. While Bacillus subtilis has inherent pathways for metabolism, there is potential enhancement via evolutionary strategies. This study aimed boost in B. using adaptive laboratory evolution (ALE) elucidate genetic changes underlying observed enhancements. The strains underwent multiple rounds (approximately 5000 generations) an environment that favored use galactose. process resulted enhanced specific growth rate 0.319 ± 0.005 h-1, a significant increase from 0.03 0.008 h-1 wild-type strains. Upon selecting evolved strain BSGA14, comprehensive whole-genome sequencing revealed presence 63 single nucleotide polymorphisms (SNPs). Two them, located coding sequences genes araR glcR, were found be advantageous mutations after reverse engineering. with these two accumulated mutations, BSGALE4, exhibited similar on BSGA14 (0.296 0.01 h-1). Furthermore, showed higher productivity protease β-galactosidase mock soybean biomass medium. ALE proved potent tool enhancing metabolism subtilis. findings offer insights into strategies microbial engineering pave way industrial applications harnessing conversion.

Language: Английский

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Fast‐track adaptive laboratory evolution of Cupriavidus necator H16 with divalent metal cations

Biotechnology Journal, Journal Year: 2024, Volume and Issue: 19(7)

Published: July 1, 2024

Abstract Microbial strain improvement through adaptive laboratory evolution (ALE) has been a key strategy in biotechnology for enhancing desired phenotypic traits. In this Biotech Method paper, we present an accelerated ALE (aALE) workflow and its successful implementation evolving Cupriavidus necator H16 enhanced tolerance toward elevated glycerol concentrations. The method involves the deliberate induction of genetic diversity controlled exposure to divalent metal cations, enabling rapid identification improved variants. Through approach, observed emergence robust variants capable growing high concentration environments, demonstrating efficacy our aALE workflow. When cultivated 10% v/v glycerol, adapted variant Mn‐C2‐B11, selected aALE, achieved final OD 600 value 56.0 dry cell weight 15.2 g L −1 , compared wild type (WT) strain's 39.1 8.4 . At even higher 15% v/v, Mn‐C2‐B11 reached 48.9 12.7 contrast WT 9.0 3.1 Higher consumption by was also confirmed high‐performance liquid chromatography (HPLC) analysis. This consumed 34.5 times more at glycerol. Our offers several advantages over other reported approaches, including independence from genetically modified strains, specialized tools, potentially carcinogenic DNA‐modifying agents. By utilizing cations as mutagens, offer safer, efficient, cost‐effective alternative expansion diversity. With ability foster microbial evolution, serves valuable addition toolbox, holding significant promise advancement engineering bioprocess optimization.

Language: Английский

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Engineering Microbial Evolution for Biotechnological Applications

RNA technologies, Journal Year: 2024, Volume and Issue: unknown, P. 133 - 161

Published: Jan. 1, 2024

Language: Английский

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Practical Achievements of Laboratory Evolution

Applied Biochemistry and Microbiology, Journal Year: 2024, Volume and Issue: 60(6), P. 1010 - 1021

Published: Dec. 1, 2024

Language: Английский

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Advances in bio-manufacturing of food functional factors based on regulating cell growth of model microorganism

Scientia Sinica Vitae, Journal Year: 2023, Volume and Issue: unknown

Published: June 29, 2023

Language: Английский

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