Rationally designed chromosome fusion does not prevent rapid growth of Vibrio natriegens

Communications Biology, Journal Year: 2024, Volume and Issue: 7(1)

Published: May 2, 2024

DNA replication is essential for the proliferation of all cells. Bacterial chromosomes are replicated bidirectionally from a single origin replication, with proceeding at about 1000 bp per second. For model organism, Escherichia coli, this translates into time 40 min its 4.6 Mb chromosome. Nevertheless, E. coli can propagate by overlapping cycles maximum short doubling 20 min. The fastest growing bacterium known, Vibrio natriegens, able to replicate generation less than 10 It has bipartite genome chromosome sizes 3.2 and 1.9 Mb. Is simultaneous two origins prerequisite rapid growth? We fused V. natriegens create strain carrying one replication. Compared parental, showed no significant deviation in growth rate. This suggests that split not growth.

Language: Английский

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Engineering xylose induction in Vibrio natriegens for biomanufacturing applications

Biotechnology and Bioengineering, Journal Year: 2024, Volume and Issue: 121(11), P. 3572 - 3581

Published: July 19, 2024

Xylose is an abundant, inexpensive and readily available carbohydrate common in minimally processed feedstocks such as seaweed algae. While a wide variety of marine microbes have evolved to utilize algae, only few currently the requisite characteristics genetic engineering tools necessary entertain use these underutilized feedstocks. The rapidly growing Gram-negative halophilic bacterium Vibrio natriegens one chassis. In this study, we engineered tested xylose induction V. tool for scalable bioproduction applications. First, created sensing construct based on operon from Escherichia coli MG1665 measured its activity using fluorescent reporter identified that cellular import plays key role strength expression required XylR transcription factor. Next, select deletions promoter region enhance gene expression, limiting effect repression when used inducer presence industrially relevant carbon sources. Lastly, optimized constructs produce biopolymer melanin seawater mimetic media. One formulations utilized nori-based extract demonstrated yields comparable previously methods more traditional costly inducer. Together, results demonstrate can provide effective lower cost option timed biosynthesis biomanufacturing applications renewable

Language: Английский

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Vibrio natriegens: Application of a Fast-Growing Halophilic Bacterium

Advances in biochemical engineering, biotechnology, Journal Year: 2024, Volume and Issue: unknown

Published: Jan. 1, 2024

Language: Английский

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Ploidy in Vibrio natriegens: Very Dynamic and Rapidly Changing Copy Numbers of Both Chromosomes

Genes, Journal Year: 2023, Volume and Issue: 14(7), P. 1437 - 1437

Published: July 13, 2023

Vibrio natriegens is the fastest-growing bacterium, with a doubling time of approximately 12-14 min. It has high potential for basic research and biotechnological applications, e.g., it can be used cell-free production (labeled) heterologous proteins, synthetic biological various compounds. However, ploidy level in V. remains unknown. At nine points throughout growth curve, we analyzed numbers origins termini both chromosomes qPCR relative abundances all genomic sites marker frequency analyses. During lag phase until early exponential growth, origin copy number origin/terminus ratio chromosome 1 increased severalfold, but increase was lower 2. This paralleled by an cell volume. phase, volume decreased again. highly dynamic fast regulation not yet been described any other species. In this study, gene dosage origin-adjacent genes during discussed together nonrandom distribution on natriegens. Taken together, results study provide first comprehensive overview dynamics will guide optimization molecular characterization applications.

Language: Английский

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Efficient Natural Plasmid Transformation ofVibrio natriegensEnables Zero-capital Molecular Biology

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Aug. 13, 2023

Abstract The fast-growing microbe Vibrio natriegens is capable of natural transformation where it draws DNA in from media via an active process under physiological conditions. Using engineered strain with a genomic copy the master competence regulator tfoX cholera combination new minimal (MCM) that uses acetate as energy source, we demonstrate naturally competent cells which are created, transformed, and recovered entirely same media, without exchange or addition media. Cells to plasmids, recombination linear DNA, co-transformation both select for scarless markerless edits. entire simple inexpensive, requiring no capital equipment room temperature (Zero Capital protocol, 10 4 cfu/ µ g), just incubator (High Efficiency 5–6 g). These retain their state when frozen transformable immediately upon thawing like typical chemical electrochemical cell. Since optimized protocol requires only 50 minutes hands-on time, V. grows quickly even on plates, started at 9 AM yields abundant culturable single colonies by 5 PM. Further, because all stages occur can be arbitrarily scaled volume, this could ideal automated directed evolution applications. As result, compete E. coli excellent chassis low-cost highly scalable synthetic biology.

Language: Английский

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