Deciphering the genetic background of an industrial 2-ketogluconic acid-producing strain Pseudomonas plecoglossicida JUIM01 using whole-genome sequencing.
Jun 2026· Biotechnology for Biofuels and Bioproducts· 0 citations
Medicine
TL;DR
The genetic determinants hypothetically linked to efficient 2KGA synthesis, including glucose metabolism, fatty acid metabolism, and the oxidative phosphorylation system are delineated, which could provide the genomic resource for elucidating high productivity and robustness, and rationally engineering the high-performance chassis cells toward robust 2KGA production.
Abstract
2-Ketogluconic acid (2KGA) is an important precursor for the food antioxidant erythorbic acid, currently produced via microbial fermentation using Pseudomonas species. To facilitate the genetic improvement of production strains, the complete genome of an industrial 2KGA producer P. plecoglossicida JUIM01 was sequenced and analyzed. The genome consists of a 5.13-Mb circular chromosome with a GC content of 63.58%, encoding 4,517 predicted proteins. Comprehensive functional annotation identified a putative global regulatory network comprising 75 core regulators, which were classified into six functionally cooperative modules, potentially governing the strain's metabolism and environmental adaptability. We further delineated the genetic determinants hypothetically linked to efficient 2KGA synthesis, including glucose metabolism, fatty acid metabolism, and the oxidative phosphorylation system. These outputs could provide the genomic resource for elucidating high productivity and robustness, and rationally engineering the high-performance chassis cells toward robust 2KGA production.
60 candidate key genes associated with high xylose-to-ethanol yield in S. stipitis are identified, predominantly involved in the cell cycle pathway, including CDC15 and PHO81.
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Metabolic engineering of Talaromyces pinophilus through promoter optimization, multicopy integration, and protease deletion enables efficient α-amylase production from lignocellulosic biomass, achieving 26 712 U/mL in bioreactor fermentation.
Jing Zeng, Jianjun Guo, Shuaiwen Zhang et al.· Journal of Industrial Microb...· 0 citations
A high-quality genome assembly and an in-depth genome analysis of V. victoriae strain D19 are presented, establishing a valuable foundation for future functional studies and providing keys for developing a new chassis for potential industrial applications.
Bartosz Wąsik, Patryk Kupaj, Paweł Moroz et al.· BMC Genomics· 0 citations
2,4-Diacetylphloroglucinol (DAPG) is a valuable antimicrobial compound with significant agricultural potential, suffers bioproduction limitations from host toxicity and inefficient downstream processing. This study engineered DAPG-hyper-tolerant Escherichia coli via adaptive laboratory evolution (ALE) starting from a phloroglucinol-tolerant strain. Optimized shake-flask fermentation of evolved Bdt03 yielded 330.52 mg/L DAPG (17.84-fold of the wild type), and the yield from the whole fermentation broth could be further increased to 391.36 mg/L. A novel organic solvent-free extraction method recovered DAPG from fermentation broth with over 98% yield via acidification, cold incubation and centrifugation, simplifying downstream processing. Genomic resequencing identified several key mutations underlying DAPG tolerance, which were validated and stacked to precisely construct strain Bb03 with enhanced production and tolerance. This work addresses the critical bottlenecks in DAPG biosynthesis by enhancing host tolerance and developing a sustainable downstream processing strategy, and also offers valuable genetic insights for constructing high-yield DAPG-producing strains and advancing the application of DAPG-responsive genetic circuits in synthetic biology.
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2'-Fucosyllactose (2'-FL), the most abundant human milk oligosaccharide (HMO), has attracted considerable interest for its prebiotic and immunomodulatory functions, with broad applications in infant nutrition. In this study, we report the development of a high-yield, genome-integrated 2'-FL-producing strain based on Escherichia coli MG1655 through systematic modular optimization. Starting from a single-copy BKHT strain (MGC06), we first optimized the copy number of the α-1,2-fucosyltransferase (α-1,2-FT) gene BKHT. Subsequently, the GDP-L-fucose supply was enhanced through coordinated genomic integration of the gene clusters cpsG-cpsB and gmd-fcl, while the multidrug efflux transporter gene mdfA was integrated to improve product export and strain robustness. BKHT copy number was then re-evaluated in the optimized background, with four copies yielding the highest production. The final engineered strain, harboring all genetic modifications stably integrated into the chromosome, produced 17.18 g/L 2'-FL in shake-flask culture. In fed-batch fermentation using a 5-L bioreactor, this strain achieved a titer of 154.12 g/L after 60 h, with a productivity of 2.57 g/L/h. Notably, throughout the entire fermentation process, no antibiotics or inducers were supplemented, underscoring the genetic stability and regulatory compliance of this plasmid-free system. To our knowledge, this represents the highest 2'-FL titer reported to date, positioning our engineered strain as a promising candidate for commercial 2'-FL production.
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