Multilevel optimisation of anaerobic ethyl acetate production in engineered Escherichia coli

Abstract Background Ethyl acetate (C4H8O2) and hydrogen (H2) are industrially relevant compounds that preferably are produced via sustainable, non-petrochemical production processes. Both compounds are volatile and can be produced by Escherichia coli before. However, relatively low yields for hydrogen are obtained and a mix of by-products renders the sole production of hydrogen by micro-organisms unfeasible. High yields for ethyl acetate have been achieved, but accumulation of formate remained an undesired but inevitable obstacle. Coupling ethyl acetate production to the conversion of formate into H2 may offer an interesting solution to both drawbacks. Ethyl acetate production requires equimolar amounts of ethanol and acetyl-CoA, which enables a redox neutral fermentation, without the need for production of by-products, other than hydrogen and CO2. Results We engineered Escherichia coli towards improved conversion of formate into H2 and CO2 by inactivating the formate hydrogen lyase repressor (hycA), both uptake hydrogenases (hyaAB, hybBC) and/or overexpressing the hydrogen formate lyase activator (fhlA), in an acetate kinase (ackA) and lactate dehydrogenase (ldhA)-deficient background strain. Initially 10 strains, with increasing number of modifications were evaluated in anaerobic serum bottles with respect to growth. Four reference strains ΔldhAΔackA, ΔldhAΔackA p3-fhlA, ΔldhAΔackAΔhycAΔhyaABΔhybBC and ΔldhAΔackAΔhycAΔhyaABΔhybBC p3-fhlA were further equipped with a plasmid carrying the heterologous ethanol acyltransferase (Eat1) from Wickerhamomyces anomalus and analyzed with respect to their ethyl acetate and hydrogen co-production capacity. Anaerobic co-production of hydrogen and ethyl acetate via Eat1 was achieved in 1.5-L pH-controlled bioreactors. The cultivation was performed at 30 °C in modified M9 medium with glucose as the sole carbon source. Anaerobic conditions and gas stripping were established by supplying N2 gas. Conclusions We showed that the engineered strains co-produced ethyl acetate and hydrogen to yields exceeding 70% of the pathway maximum for ethyl acetate and hydrogen, and propose in situ product removal via gas stripping as efficient technique to isolate the products of interest.

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Nice T-R-Y : Metabolic engineering of Escherichia coli for improved bio-based ethyl acetate production

10.18174/551681 ◽

2021 ◽

Author(s):

Anna C. Bohnenkamp

Keyword(s):

Escherichia Coli ◽

Metabolic Engineering ◽

Ethyl Acetate ◽

Acetate Production

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Study effect of crude extracts of Hibiscus sabdarifa L. on some Enterobacteriaceae

International Journal of Research in Pharmaceutical Sciences ◽

10.26452/ijrps.v9ispl1.1302 ◽

2018 ◽

Vol 9 (SPL1) ◽

Author(s):

Sabreen A Kamal ◽

Ishraq A Salih ◽

Hawraa Jawad Kadhim ◽

Zainab A Tolaifeh

Keyword(s):

Escherichia Coli ◽

Ethyl Acetate ◽

Bacterial Growth ◽

Chemical Compounds ◽

Inhibition Zone ◽

Crude Extracts ◽

And Control

Red rose or roselle (beauty rose ) is natively known as red tea belong to Malvaceae, it is flowers use traditionally for antihypertensive hepato protective, anticancer,antidiabetic,antibacterial, cytotoxicity and antidiarreal, By preparing red tea from it's flower. In this study, we extract chemical compounds by using two solvent which are Ethanol, Ethyl acetate. so we can extract Anthocyanin which is responsible for red colour of flower with many chemical compounds. then study the effect of these extracts on 5 genera from Enterobacteriacaea which can cause diarrheae (Shigella, Salmonella, Escherichia coli, Proteus and Klebsiella ) by preparing 3 concentrations for each solvent (250, 500, 750 ) mg/ml, and control then compare with two antibiotic (Azereonam 30 mg/ml and Bacitracin 10 mg/ml ) these extracts revealed obvious inhibition zone in bacterial growth.

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Faculty Opinions recommendation of High-level production of porphyrins in metabolically engineered Escherichia coli: systematic extension of a pathway assembled from overexpressed genes involved in heme biosynthesis.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1014950.195366 ◽

2003 ◽

Author(s):

Frances Arnold

Keyword(s):

Escherichia Coli ◽

Heme Biosynthesis ◽

Engineered Escherichia Coli ◽

High Level ◽

Level Production ◽

Metabolically Engineered Escherichia Coli

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Biodecomposition of Phenanthrene and Pyrene by a Genetically Engineered Escherichia coli

Recent Patents on Biotechnology ◽

10.2174/1872208314666200128103513 ◽

2020 ◽

Vol 14 (2) ◽

pp. 121-133 ◽

Cited By ~ 1

Author(s):

Maryam Ahankoub ◽

Gashtasb Mardani ◽

Payam Ghasemi-Dehkordi ◽

Ameneh Mehri-Ghahfarrokhi ◽

Abbas Doosti ◽

...

Keyword(s):

Escherichia Coli ◽

High Performance ◽

Human Cancer ◽

Genetically Engineered ◽

Hazardous Substances ◽

E Coli ◽

Spiked Soil ◽

Engineered Escherichia Coli ◽

Genetically Manipulated ◽

Hplc Measurement

Background: Genetically engineered microorganisms (GEMs) can be used for bioremediation of the biological pollutants into nonhazardous or less-hazardous substances, at lower cost. Polycyclic aromatic hydrocarbons (PAHs) are one of these contaminants that associated with a risk of human cancer development. Genetically engineered E. coli that encoded catechol 2,3- dioxygenase (C230) was created and investigated its ability to biodecomposition of phenanthrene and pyrene in spiked soil using high-performance liquid chromatography (HPLC) measurement. We revised patents documents relating to the use of GEMs for bioremediation. This approach have already been done in others studies although using other genes codifying for same catechol degradation approach. Objective: In this study, we investigated biodecomposition of phenanthrene and pyrene by a genetically engineered Escherichia coli. Methods: Briefly, following the cloning of C230 gene (nahH) into pUC18 vector and transformation into E. coli Top10F, the complementary tests, including catalase, oxidase and PCR were used as on isolated bacteria from spiked soil. Results: The results of HPLC measurement showed that in spiked soil containing engineered E. coli, biodegradation of phenanthrene and pyrene comparing to autoclaved soil that inoculated by wild type of E. coli and normal soil group with natural microbial flora, were statistically significant (p<0.05). Moreover, catalase test was positive while the oxidase tests were negative. Conclusion: These findings indicated that genetically manipulated E. coli can provide an effective clean-up process on PAH compounds and it is useful for bioremediation of environmental pollution with petrochemical products.

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