Development of a Biosensor for Crotonobetaine-CoA Ligase Screening Based on the Elucidation of Escherichia coli Carnitine Metabolism

2020 ◽  
Vol 9 (9) ◽  
pp. 2460-2471
Author(s):  
Pierre Kugler ◽  
Deborah Fröhlich ◽  
Volker F. Wendisch
Keyword(s):  
Escherichia Coli ◽  
Coa Ligase ◽  
Carnitine Metabolism

scholarly journals The fixA and fixB Genes Are Necessary for Anaerobic Carnitine Reduction in Escherichia coli

2002 ◽  
Vol 184 (14) ◽  
pp. 4044-4047 ◽  
Author(s):  
Angelique Walt ◽  
Michael L. Kahn
Keyword(s):  
Escherichia Coli ◽  
Electron Acceptor ◽  
Anaerobic Conditions ◽  
Terminal Electron Acceptor ◽  
E Coli ◽  
Carnitine Metabolism

ABSTRACT In Escherichia coli, the use of carnitine as a terminal electron acceptor depends on a functional caiTABCDE operon. It had been suggested that the adjacent but divergent fixABCX operon is also required for carnitine metabolism, perhaps to provide electrons for carnitine reduction. We have constructed E. coli fixA and fixB mutants and find that they are unable to reduce carnitine to γ-butyrobetaine under anaerobic conditions.

scholarly journals Biosynthesis of fraxetin from three different substrates using engineered Escherichia coli

2020 ◽  
Vol 63 (1) ◽  
Author(s):  
Seung Hoon An ◽  
Gyu-Sik Choi ◽  
Joong-Hoon Ahn
Keyword(s):  
Escherichia Coli ◽  
Ferulic Acid ◽  
Iron Homeostasis ◽  
Antibacterial Activities ◽  
Enzymatic Reactions ◽  
Biosynthesis Pathway ◽  
Coumaric Acid ◽  
E Coli ◽  
Coa Ligase ◽  
Key Enzyme

Abstract Fraxetin, which is a simple coumarin, is a phytochemical present in medicinal plants, such as Fraxinus rhynchophylla, and Cortex Fraxini. In plants, it serves as a controller of iron homeostasis. The health-enhancing activities of fraxetin, such as anticancer, neuroprotective and antibacterial activities, are known. Scopoletin 8-hydroxylase (S8H) is a key enzyme involved in the synthesis of fraxetin from scopoletin. Scopoletin can be synthesized either from esculetin by O-methylation or from ferulic acid by feruloyl CoA 6′-hydroxylase (F6′H) and 4-coumaric acid CoA ligase (4CL). To enable fraxetin synthesis, the fraxetin biosynthesis pathway was introduced into Escherichia coli. Three distinct routes, from ferulic acid, esculetin, and scopoletin, were designed for the synthesis of fraxetin. In the first approach, E. coli strain harboring S8H was used and found to synthesize 84.8 μM fraxetin from 100 μM scopoletin. Two E. coli strains were used for the other two approaches because these approaches required at least two enzymatic reactions. Through this approach, 41.4 μM fraxetin was synthesized from 100 μM esculetin, while 33.3 μM fraxetin was synthesized from 100 μM ferulic acid.

scholarly journals Production of 7-O-Methyl Aromadendrin, a Medicinally Valuable Flavonoid, in Escherichia coli

2011 ◽  
Vol 78 (3) ◽  
pp. 684-694 ◽  
Author(s):  
Sailesh Malla ◽  
Mattheos A. G. Koffas ◽  
Romas J. Kazlauskas ◽  
Byung-Gee Kim
Keyword(s):  
Escherichia Coli ◽  
Streptomyces Avermitilis ◽  
Flavonoid Glycosides ◽  
Petroselinum Crispum ◽  
Cell Factory ◽  
Coumaric Acid ◽  
Content Type ◽  
E Coli ◽  
Coa Ligase ◽  
Malonyl Coa

ABSTRACT7-O-Methyl aromadendrin (7-OMA) is an aglycone moiety of one of the important flavonoid-glycosides found in several plants, such asPopulus albaandEucalyptus maculata, with various medicinal applications. To produce such valuable natural flavonoids in large quantity, anEscherichia colicell factory has been developed to employ various plant biosynthetic pathways. Here, we report the generation of 7-OMA from its precursor,p-coumaric acid, inE. colifor the first time. Primarily, naringenin (NRN) (flavanone) synthesis was achieved by feedingp-coumaric acid and reconstructing the plant biosynthetic pathway by introducing the following structural genes: 4-coumarate–coenzyme A (CoA) ligase fromPetroselinum crispum, chalcone synthase fromPetunia hybrida, and chalcone isomerase fromMedicago sativa.In order to increase the availability of malonyl-CoA, a critical precursor of 7-OMA, genes for the acyl-CoA carboxylase α and β subunits (nfa9890andnfa9940), biotin ligase (nfa9950), and acetyl-CoA synthetase (nfa3550) fromNocardia farcinicawere also introduced. Thus, produced NRN was hydroxylated at position 3 by flavanone-3-hydroxylase fromArabidopsis thaliana, which was further methylated at position 7 to produce 7-OMA in the presence of 7-O-methyltransferase fromStreptomyces avermitilis. Dihydrokaempferol (DHK) (aromadendrin) and sakuranetin (SKN) were produced as intermediate products. Overexpression of the genes for flavanone biosynthesis and modification pathways, along with malonyl-CoA overproduction inE. coli, produced 2.7 mg/liter (8.9 μM) 7-OMA upon supplementation with 500 μMp-coumaric acid in 24 h, whereas the strain expressing only the flavanone modification enzymes yielded 30 mg/liter (99.2 μM) 7-OMA from 500 μM NRN in 24 h.

Thefix Escherichia coli region contains four genes related to carnitine metabolism

1995 ◽  
Vol 35 (4) ◽  
pp. 217-227 ◽  
Author(s):  
Knut Eichler ◽  
Anne Buchet ◽  
Fabienne Bourgis ◽  
Hans-Peter Kleber ◽  
Marie-Andrée Mandrand-Berthelot
Keyword(s):  
Escherichia Coli ◽  
Carnitine Metabolism

scholarly journals Biosynthesis of ethyl caffeate via caffeoyl-CoA acyltransferase expression in Escherichia coli

2021 ◽  
Vol 64 (1) ◽  
Author(s):  
Shin-Won Lee ◽  
Han Kim ◽  
Joong-Hoon Ahn
Keyword(s):  
Escherichia Coli ◽  
Caffeic Acid ◽  
Shikimate Pathway ◽  
Acid Synthesis ◽  
Biological Properties ◽  
E Coli ◽  
Pathway Gene ◽  
Coa Ligase ◽  
Prunus Yedoensis ◽  
Tyrosine Ammonia Lyase

AbstractHydroxycinnamic acids (HCs) are natural compounds that form conjugates with diverse compounds in nature. Ethyl caffeate (EC) is a conjugate of caffeic acid (an HC) and ethanol. It has been found in several plants, including Prunus yedoensis, Polygonum amplexicaule, and Ligularia fischeri. Although it exhibits anticancer, anti-inflammatory, and antifibrotic activities, its biosynthetic pathway in plants still remains unknown. This study aimed to design an EC synthesis pathway and clone genes relevant to the same. Genes involved in the caffeic acid synthesis pathway (tyrosine ammonia-lyase (TAL) and p-coumaric acid hydroxylase (HpaBC)) were introduced into Escherichia coli along with 4-coumaroyl CoA ligase (4CL) and acyltransferases (AtCAT) cloned from Arabidopsis thaliana. In presence of ethanol, E. coli harboring the above genes successfully synthesized EC. Providing more tyrosine through the overexpression of shikimate-pathway gene-module construct and using E. coli mutant enhanced EC yield; approximately 116.7 mg/L EC could be synthesized in the process. Synthesis of four more alkyl caffeates was confirmed in this study; these might potentially possess novel biological properties, which would require further investigation.

scholarly journals Model Driven Analysis of the Biosynthesis of 1,4-butanediol from Renewable Feedstocks in Escherichia coli

2019 ◽  
Vol 70 (11) ◽  
pp. 3808-3817
Author(s):  
Zsolt Bodor ◽  
Szabolcs Lanyi ◽  
Beata Albert ◽  
Katalin Bodor ◽  
Aurelia Cristina Nechifor ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Metabolic Model ◽  
E Coli ◽  
Model Driven ◽  
Metabolic Characteristics ◽  
Strain Design ◽  
Renewable Feedstocks ◽  
Coa Ligase ◽  
Coa Transferase ◽  
Genome Scale

Bio-based, environmentally benign production of commodity chemicals such as 1,4-butanediol (BDO) from renewable feedstocks is highly challenging due to the lack of natural synthesis pathways. Herein, we present a systematic model-driven evaluation of the production potential for Escherichia coli to produce BDO from renewable carbohydrates (glucose, glycerol). Computational analysis was carried out in order to decipher the metabolic characteristics under various genetic and environmental conditions. Optimal strain designs were achieved using only two (adhE2- alcohol dehydrogenase and cat/sucCD- 4-hydroxybutyrate-CoA transferase/4-hydroxybutyryl-CoA ligase) heterologous reactions; highest yields were attained for: glucose ~0.37 g g-1 (3 knockouts, anaerobically) and glycerol ~0.43 g g-1 (4 knockouts, microaerobically). The maximum achievable production yield was over 95% of the theoretical maximum potential for glucose and over 75% for glycerol. In regards to the genome-scale metabolic model predictions, a metabolically engineered E. coli was created to analyze the new biosynthetic pathway stability and functionality. Considering the preliminary outcomes the strain and pathway is stable under fermentative conditions and a limited quantity of BDO ~1 mg L-1 was obtained, therefore long-term adaptive evolution is mandatory. This study outlines a strain design and analysis pipeline -systems biology-based approach- for non-native compounds production strains.

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