Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Oct;43(10):2035-2043.
doi: 10.1007/s10529-021-03169-z. Epub 2021 Aug 26.

Modification of an engineered Escherichia coli by a combinatorial strategy to improve 3,4-dihydroxybutyric acid production

Yidi Liu  1 , Xinlei Mao  1 , Baoqi Zhang  1 , Jinping Lin  2 , Dongzhi Wei  1
Affiliations

Modification of an engineered Escherichia coli by a combinatorial strategy to improve 3,4-dihydroxybutyric acid production

Yidi Liu et al. Biotechnol Lett. 2021 Oct.

Abstract

Objectives: 3,4-Dihydroxybutyric acid (3,4-DHBA) is a multifunctional C4 platform compound widely used for the synthesis of various materials, including pharmaceuticals. Although, a biosynthetic pathway for 3,4-DHBA production has been developed, its low yield still precludes large-scale use. Here, a heterologous four-step biosynthetic pathway was established in recombinant Escherichia coli (E. coli) using a combinatorial strategy.

Results: Several aldehyde dehydrogenases (ALDHs) were screened, using in vitro enzyme assays, to identify suitable catalysts for the dehydrogenation of 3,4-dihydroxybutanal (3,4-DHB) to 3,4-DHBA. A pathway containing glucose dehydrogenase (BsGDH) from Bacillus subtilis, D-xylonate dehydratase (YagF) from E. coli, benzoylformate decarboxylase (PpMdlC) from Pseudomonas putida and ALDH was introduced into E. coli, generating 3.04 g/L 3,4-DHBA from D-xylose (0.190 g 3,4-DHBA/g D-xylose). Disruption of competing pathways by deleting xylA, ghrA, ghrB and adhP contributed to an 87% increase in 3,4-DHBA accumulation. Expression of a fusion construct containing PpMdlC and YagF enhanced the 3,4-DHBA titer, producing the highest titer and yield reported thus far (7.71 g/L; 0.482 g 3,4-DHBA/g D-xylose).

Conclusions: These results showed that deleting genes from competing pathways and constructing fusion proteins significantly improved the titer and yield of 3,4-DHBA in engineered E. coli.

Keywords: 3,4-DHBA; Competing pathway; D-Xylose; E. coli; Fusion protein.

PubMed Disclaimer

References

    1. Albertsen L, Chen Y, Bach LS, Rattleff S, Maury J, Brix S, Nielsen J, Mortensen UH (2011) Diversion of flux toward sesquiterpene production in Saccharomyces cerevisiae by fusion of host and heterologous enzymes. Appl Environ Microbiol 77:1033–1040 - DOI
    1. Brower PL, Butler DE, Deering CF, Le TV, Millar A, Nanninga TN, Roth D (1992) The synthesis of (4R-cis)-1,1-dimethylethyl 6-cyanomethyl-2,2-dimethyl-1,3-dioxane-4-acetate, a key intermediate for the preparation of CI-981, a highly potent, tissue selective inhibitor of HMG-CoA reductase. Tetrahedron Lett 33:2279–2282 - DOI
    1. Chen K, Li K, Deng J, Zhang B, Lin J, Wei D (2016) Carbonyl reductase identification and development of whole-cell biotransformation for highly efficient synthesis of (R)-[3,5-bis(trifluoromethyl)phenyl] ethanol. Microb Cell Fact 15:191 - DOI
    1. Choi KY, Wernick DG, Tat CA, Liao JC (2014) Consolidated conversion of protein waste into biofuels and ammonia using Bacillus subtilis. Metab Eng 23:53–61 - DOI
    1. Choi S, Song CW, Shin JH, Lee SY (2015) Biorefineries for the production of top building block chemicals and their derivatives. Metab Eng 28:223–239 - DOI

MeSH terms

LinkOut - more resources