Microbial upgrading of acetate into 2,3-butanediol and acetoin by E. coli W DOI Creative Commons
Katharina Novak, Regina Kutscha, Stefan Pflügl

et al.

Biotechnology for Biofuels, Journal Year: 2020, Volume and Issue: 13(1)

Published: Oct. 22, 2020

Abstract Background Acetate is an abundant carbon source and its use as alternative feedstock has great potential for the production of fuel platform chemicals. Acetoin 2,3-butanediol represent two these Results The aim this study was to produce acetoin from acetate in Escherichia coli W. key strategies achieve goal were: strain engineering, detail deletion mixed-acid fermentation pathways E. W Δ ldhA adhE pta frdA 445_Ediss development a new defined medium containing five amino acids seven vitamins. Stepwise reduction media additives further revealed that diol mediated by availability aspartate. Other or TCA cycle intermediates did not enable growth on acetate. Cultivation under controlled conditions batch pulsed fed-batch experiments showed aspartate consumed before acetate, indicating co-utilization prerequisite production. addition gave cultures start-kick required feeding. Pulsed fed-batches resulted 1.43 g l −1 1.16 diols (2,3-butanediol acetoin) alone. yield reached 0.09 per which accounts 26% theoretical maximum. Conclusion This first time well chemically product formation . Hereby, we provide solid base process intensification investigation other products.

Language: Английский

Prospects on bio-based 2,3-butanediol and acetoin production: Recent progress and advances DOI
Sofia Maina, Ashish A. Prabhu, Vivek Narisetty

et al.

Biotechnology Advances, Journal Year: 2021, Volume and Issue: 54, P. 107783 - 107783

Published: June 29, 2021

Language: Английский

Citations

123
Advances in developing metabolically engineered microbial platforms to produce fourth-generation biofuels and high-value biochemicals DOI
Muhammad Aamer Mehmood, Ayesha Shahid, Sana Malik

et al.

Bioresource Technology, Journal Year: 2021, Volume and Issue: 337, P. 125510 - 125510

Published: July 7, 2021

Language: Английский

Citations

57
Metabolic engineering of Vibrio natriegens DOI Creative Commons
Felix Thoma, Bastian Blombach

Essays in Biochemistry, Journal Year: 2021, Volume and Issue: 65(2), P. 381 - 392

Published: April 9, 2021

Vibrio natriegens is emerging as a promising host for biotechnology which basically due to the remarkable intrinsic properties such exceptionally high growth and substrate consumption rates. The facultatively anaerobic marine bacterium possesses versatile metabolism, able utilize variety of substrates carbon energy sources easy handle in lab. These features initiated rapid development genetic tools resulted extensive engineering production strains past years. Although recent examples illustrate potential V. biotechnology, comprehensive understanding metabolism its regulation still lacking but essential exploit full this bacterium. In review, we summarize current knowledge on physiological traits genomic organization, provide an overview available advances metabolic natriegens. Finally, discuss obstacles have be overcome order establish industrial host.

Language: Английский

Citations

51
The Marburg Collection: A Golden Gate DNA Assembly Framework for Synthetic Biology Applications in Vibrio natriegens DOI
Daniel Stukenberg,

Tobias Hensel,

Josef Hoff

et al.

ACS Synthetic Biology, Journal Year: 2021, Volume and Issue: 10(8), P. 1904 - 1919

Published: July 13, 2021

Vibrio natriegens is known as the world's fastest growing organism with a doubling time of less than 10 min. This incredible growth speed empowers V. chassis for synthetic and molecular biology, potentially replacing E. coli in many applications. While first genetic parts have been built tested natriegens, comprehensive toolkit containing well-characterized standardized did not exist. To close this gap, we created Marburg Collection—a highly flexible Golden Gate cloning toolbox optimized emerging 191 parts. The Collection overcomes paradigm plasmid construction—integrating inserts into backbone—by enabling de novo assembly plasmids from basic allows users to select replication origin resistance part independently, which advantageous when limited knowledge about behavior those target available. Additional design highlights are novel connector parts, facilitate modular circuit and, optionally, inversion individual transcription units reduce transcriptional crosstalk multigene constructs. quantitatively characterize contained developed reliable microplate reader measurement workflow reporter experiments overcame organism-specific challenges. We think its thoroughly characterized will provide valuable resource community.

Language: Английский

Citations

49
Efficient natural plasmid transformation of Vibrio natriegens enables zero-capital molecular biology DOI Creative Commons
David A. Specht, Timothy J. Sheppard, Finn Kennedy

et al.

PNAS Nexus, Journal Year: 2024, Volume and Issue: 3(2)

Published: Feb. 1, 2024

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 cholerae 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 fresh media. Cells to plasmids, recombination linear DNA, cotransformation both select for scarless markerless edits. entire simple inexpensive, requiring no capital equipment room temperature (zero protocol, 104 cfu/μg), just incubator (high-efficiency 105−6 cfu/μg). These retain their state when frozen transformable immediately upon thawing like typical chemical electrochemical cell. Since optimized protocol requires only 50 min 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 Escherichia coli excellent chassis low-cost highly scalable synthetic biology.

Language: Английский

Citations

7
Combinatorial metabolic pathway assembly approaches and toolkits for modular assembly DOI
Rosanna Young, Matthew Haines, Marko Storch

et al.

Metabolic Engineering, Journal Year: 2020, Volume and Issue: 63, P. 81 - 101

Published: Dec. 7, 2020

Language: Английский

Citations

43
Vibrio natriegens as a pET-Compatible Expression Host Complementary to Escherichia coli DOI Creative Commons
Jiaqi Xu, Feng Dong,

Meixian Wu

et al.

Frontiers in Microbiology, Journal Year: 2021, Volume and Issue: 12

Published: Feb. 19, 2021

Efficient and novel recombinant protein expression systems can further reduce the production cost of enzymes. Vibrio natriegens is fastest growing free-living bacterium with a doubling time less than 10 min, which makes it highly attractive as host. Here, 196 pET plasmids different genes interest (GOIs) were electroporated into V. strain VnDX, carries an integrated T7 RNA polymerase cassette. As result, 65 75% tested GOIs obtained soluble in Escherichia coli , respectively, 20 showed better former. Furthermore, we have adapted consensus “what to try first” protocol for based on Terrific Broth medium. Six sampled encoding biocatalysts enzymes thus achieved 50–128% higher catalytic efficiency under optimized conditions. Our study demonstrated pET-compatible host spectrum expressed distinct from E. easy-to-use protocol, solving problem that some cannot be well .

Language: Английский

Citations

39
Non-Sterilized Fermentation of 2,3-Butanediol with Seawater by Metabolic Engineered Fast-Growing Vibrio natriegens DOI Creative Commons
Wensi Meng, Yongjia Zhang, Liting Ma

et al.

Frontiers in Bioengineering and Biotechnology, Journal Year: 2022, Volume and Issue: 10

Published: July 12, 2022

Sustainable and environment-friendly microbial fermentation processes have been developed to produce numerous chemicals. However, the high energy input required for sterilization substantial fresh water consumption restrict economic feasibility of traditional processes. To address these problems, Vibrio natriegens, a promising chassis with low nutritional requirements, salt tolerance rapid growth rate can be selected as host chemical production. In this study, V. natriegens was metabolic engineered 2,3-butanediol (2,3-BD), an important platform chemical, through non-sterilized seawater-based minimal medium after expressing 2,3-BD synthesis cluster deleting two byproduct encoding genes. Under optimized fermentative conditions, 41.27 g/L produced productivity 3.44 g/L/h yield 0.39 g/g glucose by recombinant strain natriegensΔfrdAΔldhA-pETRABC. This study confirmed using seawater replace freshwater other valuable chemicals may also engineering emerging synthetic biology natriegens.

Language: Английский

Citations

28
Systems biology of competency in Vibrio natriegens is revealed by applying novel data analytics to the transcriptome DOI Creative Commons
Jongoh Shin, Kevin Rychel, Bernhard Ø. Palsson

et al.

Cell Reports, Journal Year: 2023, Volume and Issue: 42(6), P. 112619 - 112619

Published: June 1, 2023

Vibrio natriegens regulates natural competence through the TfoX and QstR transcription factors, which are involved in external DNA capture transport. However, extensive genetic transcriptional regulatory basis for competency remains unknown. We used a machine-learning approach to decompose natriegens's transcriptome into 45 groups of independently modulated sets genes (iModulons). Our findings show that is associated with repression two housekeeping iModulons (iron metabolism translation) activation six iModulons; including QstR, novel iModulon unknown function, three (representing motility, polycations, reactive oxygen species [ROS] responses). Phenotypic screening 83 gene deletion strains demonstrates loss function reduces or eliminates competency. This database-iModulon-discovery cycle unveils transcriptomic its relationship functions. These results provide systems biology this organism.

Language: Английский

Citations

17
Metabolic engineering of Vibrio natriegens for anaerobic succinate production DOI
Felix Thoma, Clarissa Schulze,

Carolina Gutierrez‐Coto

et al.

Microbial Biotechnology, Journal Year: 2021, Volume and Issue: 15(6), P. 1671 - 1684

Published: Nov. 29, 2021

The biotechnological production of succinate bears serious potential to fully replace existing petrochemical approaches in the future. In order establish an economically viable bioprocess, obtaining high titre, yield and productivity is central importance. this study, we present a straightforward engineering approach for anaerobic with Vibrio natriegens, consisting essential metabolic optimization process conditions. final producer strain V. natriegens Δlldh Δdldh Δpfl Δald Δdns::pycCg (Succ1) yielded 1.46 mol per glucose under conditions (85% theoretical maximum) revealed particularly biomass-specific rate 1.33 gSucc gCDW-1 h-1 compared well-established systems. By applying carbon redox balancing, determined intracellular flux distribution show that tested reductive TCA as well oxidative TCA/glyoxylate pathway contributed formation. zero-growth bioprocess using minimal medium devoid complex additives expensive supplements, obtained titre 60.4 l-1 maximum 20.8 overall volumetric 8.6 during 7 h fermentation. key performance indicators (titre, productivity) first are encouraging compete costly tailored microbial

Language: Английский

Citations

28