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

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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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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Metabolic Engineering of Saccharomyces cerevisiae for Ethyl Acetate Biosynthesis

ACS Synthetic Biology ◽

10.1021/acssynbio.0c00446 ◽

2021 ◽

Author(s):

Wenqi Shi ◽

Jie Li ◽

Yanfang Chen ◽

Xiaohang Liu ◽

Yefu Chen ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Metabolic Engineering ◽

Ethyl Acetate

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Metabolic Engineering of Escherichia coli for Lacto-N-triose II Production with High Productivity

Journal of Agricultural and Food Chemistry ◽

10.1021/acs.jafc.1c00246 ◽

2021 ◽

Author(s):

Yingying Zhu ◽

Li Wan ◽

Jiawei Meng ◽

Guocong Luo ◽

Geng Chen ◽

...

Keyword(s):

Escherichia Coli ◽

Metabolic Engineering ◽

High Productivity

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Improving Ethyl Acetate Production in Baijiu Manufacture by Wickerhamomyces anomalus and Saccharomyces cerevisiae Mixed Culture Fermentations

BioMed Research International ◽

10.1155/2019/1470543 ◽

2019 ◽

Vol 2019 ◽

pp. 1-11 ◽

Cited By ~ 4

Author(s):

Guangsen Fan ◽

Chao Teng ◽

Dai Xu ◽

Zhilei Fu ◽

Pengxiao Liu ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Ethyl Acetate ◽

Mixed Culture ◽

Nutrient Competition ◽

Acetate Production ◽

Mixed Fermentation ◽

Wickerhamomyces Anomalus ◽

Flavor Perception ◽

Positive Effect

Ethyl acetate content has strong influence on the style and quality of Baijiu. Therefore, this study investigated the effect of Saccharomyces cerevisiae Y3401 on the production of ethyl acetate by Wickerhamomyces anomalus Y3604. Analysis of cell growth showed that Y3401 influences Y3604 by nutrient competition and inhibition by metabolites, while the effect of Y3604 on Y3401 was mainly competition for nutrients. Mixed fermentation with two yeasts was found to produce more ethyl acetate than a single fermentation. The highest yield of ethyl acetate was 2.99 g/L when the inoculation ratio of Y3401:Y3604 was 1:2. Synergistic fermentation of both yeasts improved ethyl acetate production and increased the content of other flavor compounds in liquid and simulated solid-state fermentation for Baijiu. Saccharomyces cerevisiae had a positive effect on ethyl acetate production in mixed culture and provides opportunities to alter the aroma and flavor perception of Baijiu.

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