Characterization of acetyl-CoA synthetase kinetics and ATP-binding

Gallego‐Jara, Julia; Terol, Gema Lozano; Conesa, Ana Écija; Zambelli, Barbara; Díaz, Manuel Cánovas; Diego, Teresa De

doi:10.1016/j.bbagen.2019.03.017

Cited by 14 publications

(13 citation statements)

References 58 publications

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“…We also detected a decrease in OD 600 , presumably due to a toxic effect of p -coumaric acid accumulation in cells. In addition, we also did not observe higher scutellarin production either by overexpressing SeACS P641L [ 35 , 51 ] and PEX10 [ 49 ], which could increase the malonyl-CoA pool ( Fig. S4 ).…”

Section: Resultsmentioning

confidence: 92%

Metabolic engineering of Yarrowia lipolytica for scutellarin production

Wang

Liu

Chen

et al. 2022

Synthetic and Systems Biotechnology

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Section: Resultsmentioning

confidence: 92%

Metabolic engineering of Yarrowia lipolytica for scutellarin production

Wang

Liu

Chen

et al. 2022

Synthetic and Systems Biotechnology

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“…Heterologous expression of the Salmonella enterica acetyl‐CoA synthetase mutant (Acs L641P ), [28] in which the critical residue L641 for recognition by the Lys‐A10 acetyltransferase Pat is mutated, and engineering acetyl‐CoA synthetases in corresponding residue are the most widely used approaches to gain the immunity of Lys‐A10 acylation [29] . However, mutation of L641 will significantly reduce the catalytic activity of acetyl‐CoA synthetases, [30] e.g. Acs L641P is only about one‐third that of the wildtype enzyme [7c] .…”

Section: Resultsmentioning

confidence: 99%

An Atypical Acyl‐CoA Synthetase Enables Efficient Biosynthesis of Extender Units for Engineering a Polyketide Carbon Scaffold

Zheng

Zhang

et al. 2022

Angewandte Chemie

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Acyl-CoAs are key precursors of primary and secondary metabolism. Their efficient biosynthesis is often impeded by the limited substrate specificity and low in vivo activity of acyl-CoA synthetases (ACSs) due to regulatory acylation of the catalytically important lysine residue in motif A10 (Lys-A10). In this study, we identified an unusual ACS (UkaQ) from the UK-2A biosynthetic pathway that naturally lacks the Lys-A10 residue and exhibits extraordinarily broad substrate specificity. Protein engineering significantly improved its stability and catalytic activity, enabling it to synthesize a large variety of acyl-CoAs with highly robust activity. By combining it with permissive carboxylases, we produced a large array of polyketide extender units and obtained six novel halobenzyl-containing antimycin analogues through an engineered biosynthetic pathway. This study significantly expands the catalytic mode of ACSs and provides a potent tool for the biosynthesis of acyl-CoA-derived natural products.

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Section: Resultsmentioning

confidence: 99%

“…Acetyl-CoA synthase (ACS) is a member of the superfamily of adenylating enzymes, which can convert intracellular acetic acid to acetyl-CoA, the precursor for fatty acid synthesis. 29 To further enhance the acetate utilization capability, acs was first overexpressed in Y. lipolytica PO1f. Five ACS genes from different resources including Y. lipolytica, Saitozyma podzolica, Rhodosporidium toruloides, Saccharomyces cerevisiae, and Salmonella enterica were selected and compared.…”

Section: Resultsmentioning

confidence: 99%

“…Compared with the ACS of S. cerevisiae, valine at position 585 was changed to isoleucine in acs SP , and histidine at the 650 position was changed to glutamic acid in acs SP , which has been reported to affect the ATP recognition and substrate acetylation. 29 The differences in key sites will affect the expression of the enzyme. The changes in the key sites of ACS RT are exactly the same as ACS SP , but the final modification effect was not as good as ACS SP .…”

Section: Resultsmentioning

confidence: 99%

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Increased Lipid Production in Yarrowia lipolytica from Acetate through Metabolic Engineering and Cosubstrate Fermentation

et al. 2021

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Bioconversion of acetate, a byproduct generated in industrial processes, into microbial lipids using oleaginous yeasts offers a promising alternative for the economic utilization of acetate-containing waste streams. However, high acetate concentrations will inhibit microbial growth and metabolism. In this study, the acetate utilization capability of Yarrowia lipolytica PO1f was successively improved by overexpressing the key enzyme of acetyl-CoA synthetase (ACS), which resulted in an accumulation of 9.2% microbial lipids from acetate in shake flask fermentation. By further overexpressing the second key enzymes of acetyl-CoA carboxylase (ACC1) and fatty acid synthase (FAS) in Y. lipolytica, the lipid content was increased to 25.7% from acetate. Finally, the maximum OD600 of 29.2 and a lipid content of 41.7% were obtained with the engineered strain by the adoption of cosubstrate (glycerol and acetate) fed-batch fermentation, which corresponded to an increase of 68 and 95%, respectively. These results presented a promising strategy for economic and efficient microbial lipid production from the waste acetate.

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Characterization of acetyl-CoA synthetase kinetics and ATP-binding

Cited by 14 publications

References 58 publications

Metabolic engineering of Yarrowia lipolytica for scutellarin production

Metabolic engineering of Yarrowia lipolytica for scutellarin production

An Atypical Acyl‐CoA Synthetase Enables Efficient Biosynthesis of Extender Units for Engineering a Polyketide Carbon Scaffold

Increased Lipid Production in Yarrowia lipolytica from Acetate through Metabolic Engineering and Cosubstrate Fermentation

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