Improved production of propionic acid using genome shuffling

Luna‐Flores, Carlos H.; Palfreyman, Robin W.; Krömer, Jens O.; Nielsen, Lars K.; Marcellin, Esteban

doi:10.1002/biot.201600120

Cited by 27 publications

(24 citation statements)

References 61 publications

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“…Furthermore, utilization of lowcost cane molasses by P. acidipropionici action could generate 30 g/L propionic acid [32]. In addition, genome shuffling technology could increase 25% of P. acidipropionici [33]. Moreover, the production of propionic acid reached at 26.95 g/L and 34.93 g/L by the action of engineered P. jenseniis [34].…”

Section: Microbial Production Of Particular Vfas and Hydrogen Using Pmentioning

confidence: 99%

Bioengineering of anaerobic digestion for volatile fatty acids, hydrogen or methane production: A critical review

et al. 2019

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Anaerobic digestion (AD) is a well-established technology used for producing biogas or biomethane alongside the slurry used as biofertilizer. However, using a variety of wastes and residuals as substrate and mixed cultures in the bioreactor makes AD as one of the most complicated biochemical processes employing hydrolytic, acidogenic, hydrogen-producing, acetate-forming bacteria as well as acetoclastic and hydrogenoclastic methanogens. Hydrogen and volatile fatty acids (VFAs) including acetic, propionic, isobutyric, butyric, isovaleric, valeric and caproic acid and other carboxylic acids such as succinic and lactic acids are formed as intermediate products. As these acids are important precursors for various industries as mixed or purified chemicals, the AD process can be bioengineered to produce VFAs alongside hydrogen and therefore biogas plants can become biorefineries. The current review paper provides the theory and means to produce and accumulate VFAs and hydrogen, inhibit their conversion to methane and to extract them as the final products. The effects of pretreatment, pH, temperature, hydraulic retention time (HRT), organic loading rate (OLR), chemical methane inhibitions, and heat shocking of the inoculum on VFAs accumulation, hydrogen production, VFAs composition, and the microbial community were discussed. Furthermore, this paper highlights the possible techniques for recovery of VFAs from the fermentation media in order to minimize product inhibition as well as to supply the carboxylates for downstream procedures.

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Section: Microbial Production Of Particular Vfas and Hydrogen Using Pmentioning

confidence: 99%

Bioengineering of anaerobic digestion for volatile fatty acids, hydrogen or methane production: A critical review

et al. 2019

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“…Given limitations in the availability of tools for rational strain design, random mutagenesis and/or adaptive evolution have been the best strategies to improve the yield of PA in propionibacterium . We have previously used genome shuffling (GS) to obtain a recombinant strain capable of producing PA at a yield of 0.55 g/g which was close to the target yield of 0.6 g/g [ 5 – 7 ]. Developed in the early 2000s, GS is a rapid phenotypic improvement technique based on protoplast fusion [ 8 , 9 ].…”

Section: Introductionmentioning

confidence: 99%

Linking genotype and phenotype in an economically viable propionic acid biosynthesis process

Luna‐Flores

Stowers²,

Cox³

et al. 2018

Biotechnol Biofuels

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BackgroundPropionic acid (PA) is used as a food preservative and increasingly, as a precursor for the synthesis of monomers. PA is produced mainly through hydrocarboxylation of ethylene, also known as the ‘oxo-process’; however, Propionibacterium species are promising biological PA producers natively producing PA as their main fermentation product. However, for fermentation to be competitive, a PA yield of at least 0.6 g/g is required.ResultsA new strain able to reach the required yield was obtained using genome shuffling. To gain insight into the changes leading to the improved phenotype, the genome of the new strain was sequenced, and metabolomics and transcriptomics data were obtained. In combination, the data revealed three key mutations: (i) a mutation in the promoter region of a sugar transporter which enables an increase in the uptake rate of sucrose; (ii) a mutation in a polar amino acid transporter which improves consumption of amino acids and acid tolerance; and (iii) a mutation in a gene annotated as a cytochrome C biogenesis gene, which is likely responsible for the coupling of the Wood–Werkman cycle to ATP production were responsible for the phenotype. The bioprocess was further enhanced with a feeding strategy that achieved 70 g/L of product. The proposed bioprocess is expected to outperform the economics of the current ‘oxo-process’ by 2020.ConclusionsIn this study, using genome shuffling, we obtained a strain capable of producing PA exceeding the commercial needs. The multiomics comparison between the novel strain and the wild type revealed overexpression of amino acid pathways, changes in sucrose transporters and an increased activity in the methylglyoxal and the glucuronate interconversion pathways. The analysis also suggests that a mutation in the cytochrome C biogenesis gene, coupled with ATP production through the Wood–Werkman cycle, may be responsible for the increased PA production.Electronic supplementary materialThe online version of this article (10.1186/s13068-018-1222-9) contains supplementary material, which is available to authorized users.

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“…Genome shuffling in propionibacteria has been used to improve vitamin B12 production in P. shermanii [78] and propionic acid in P. acidipropionici [71,75,76]. Recently, multiple propionibacteria species were genome shuffled resulting in a strain which achieved a yield of 0.55 g propionate/g glucose [75,76].…”

Section: Empirical Strain Designmentioning

confidence: 99%

“…One option is relying on high-throughput HPLC to identify high producers [6]. Alternatively, growth rate may be used to identify mutants that are more acid tolerant and exhibit improved propionate yields [10,75,76].…”

Section: Empirical Strain Designmentioning

confidence: 99%

“…Recently, multiple propionibacteria species were genome shuffled resulting in a strain which achieved a yield of 0.55 g propionate/g glucose [75,76]. Next-generation sequencing was used to analyse recombination events and identify novel/unique regions from each strain leading to the improved phenotype, including changes linked to acid tolerance mechanisms and possibly to a new transcriptional mechanism through mutation in ribosomal RNAs.…”

Section: Empirical Strain Designmentioning

confidence: 99%

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Microbial Propionic Acid Production

et al. 2017

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Propionic acid (propionate) is a commercially valuable carboxylic acid produced through microbial fermentation. Propionic acid is mainly used in the food industry but has recently found applications in the cosmetic, plastics and pharmaceutical industries. Propionate can be produced via various metabolic pathways, which can be classified into three major groups: fermentative pathways, biosynthetic pathways, and amino acid catabolic pathways. The current review provides an in-depth description of the major metabolic routes for propionate production from an energy optimization perspective. Biological propionate production is limited by high downstream purification costs which can be addressed if the target yield, productivity and titre can be achieved. Genome shuffling combined with high throughput omics and metabolic engineering is providing new opportunities, and biological propionate production is likely to enter the market in the not so distant future. In order to realise the full potential of metabolic engineering and heterologous expression, however, a greater understanding of metabolic capabilities of the native producers, the fittest producers, is required.

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Improved production of propionic acid using genome shuffling

Cited by 27 publications

References 61 publications

Bioengineering of anaerobic digestion for volatile fatty acids, hydrogen or methane production: A critical review

Bioengineering of anaerobic digestion for volatile fatty acids, hydrogen or methane production: A critical review

Linking genotype and phenotype in an economically viable propionic acid biosynthesis process

Microbial Propionic Acid Production

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