scholarly journals Improving Production of Malonyl Coenzyme A-Derived Metabolites by Abolishing Snf1-Dependent Regulation of Acc1

mBio ◽  
2014 ◽  
Vol 5 (3) ◽  
Author(s):  
Shuobo Shi ◽  
Yun Chen ◽  
Verena Siewers ◽  
Jens Nielsen
Keyword(s):  
Fatty Acid ◽  
Coenzyme A ◽  
Fatty Acid Ethyl Esters ◽  
Ethyl Esters ◽  
Regulation Of Transcription ◽  
Acetyl Coa ◽  
Content Type ◽  
Obesity And Diabetes ◽  
Malonyl Coa ◽  
Hydroxypropionic Acid

ABSTRACT Acetyl coenzyme A (acetyl-CoA) carboxylase (ACCase) plays a central role in carbon metabolism and has been the site of action for the development of therapeutics or herbicides, as its product, malonyl-CoA, is a precursor for production of fatty acids and other compounds. Control of Acc1 activity in the yeast Saccharomyces cerevisiae occurs mainly at two levels, i.e., regulation of transcription and repression by Snf1 protein kinase at the protein level. Here, we demonstrate a strategy for improving the activity of ACCase in S. cerevisiae by abolishing posttranslational regulation of Acc1 via site-directed mutagenesis. It was found that introduction of two site mutations in Acc1, Ser659 and Ser1157, resulted in an enhanced activity of Acc1 and increased total fatty acid content. As Snf1 regulation of Acc1 is particularly active under glucose-limited conditions, we evaluated the effect of the two site mutations in chemostat cultures. Finally, we showed that our modifications of Acc1 could enhance the supply of malonyl-CoA and therefore successfully increase the production of two industrially important products derived from malonyl-CoA, fatty acid ethyl esters and 3-hydroxypropionic acid. IMPORTANCE ACCase is responsible for carboxylation of acetyl-CoA to produce malonyl-CoA, which is a crucial step in the control of fatty acid metabolism. ACCase opened the door for pharmaceutical treatments of obesity and diabetes as well as the development of new herbicides. ACCase is also recognized as a promising target for developing cell factories, as its malonyl-CoA product serves as a universal precursor for a variety of high-value compounds in white biotechnology. Yeast ACCase is a good model in understanding the enzyme’s catalysis, regulation, and inhibition. The present study describes the importance of protein phosphorylation in regulation of yeast ACCase and identifies potential regulation sites. This study led to the generation of a more efficient ACCase, which was applied in the production of two high-value compounds derived from malonyl-CoA, i.e., fatty acid ethyl esters that can be used as biodiesel and 3-hydroxypropionic acid that is considered an important platform chemical.

The Classical, Yet Controversial, First Enzyme of Lipid Synthesis: Escherichia coli Acetyl-CoA Carboxylase

2021 ◽  
Author(s):  
John E. Cronan
Keyword(s):  
Escherichia Coli ◽  
Fatty Acid ◽  
Fatty Acid Synthesis ◽  
Building Block ◽  
Lipid Synthesis ◽  
Acid Synthesis ◽  
Acetyl Coa Carboxylase ◽  
Acetyl Coa ◽  
Content Type ◽  
Malonyl Coa

SUMMARY Escherichia coli acetyl-CoA carboxylase (ACC), the enzyme responsible for synthesis of the malonyl-CoA, the building block of fatty acid synthesis, is the paradigm bacterial ACC. Many reports on the structures and stoichiometry of the four subunits comprising the active enzyme as well as on regulation of ACC activity and expression have appeared in the almost 20 years since this subject was last reviewed. This review seeks to update and expand on these reports.

scholarly journals Structure, Activity, and Inhibition of the Carboxyltransferase β-Subunit of Acetyl Coenzyme A Carboxylase (AccD6) from Mycobacterium tuberculosis

2014 ◽  
Vol 58 (10) ◽  
pp. 6122-6132 ◽  
Author(s):  
Manchi C. M. Reddy ◽  
Ardala Breda ◽  
John B. Bruning ◽  
Mukul Sherekar ◽  
Spandana Valluru ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Fatty Acid Biosynthesis ◽  
Coenzyme A ◽  
Cell Activity ◽  
Acetyl Coa ◽  
Acetyl Coenzyme A ◽  
Content Type ◽  
Acetyl Coenzyme ◽  
Malonyl Coa

ABSTRACTInMycobacterium tuberculosis, the carboxylation of acetyl coenzyme A (acetyl-CoA) to produce malonyl-CoA, a building block in long-chain fatty acid biosynthesis, is catalyzed by two enzymes working sequentially: a biotin carboxylase (AccA) and a carboxyltransferase (AccD). While the exact roles of the three different biotin carboxylases (AccA1 to -3) and the six carboxyltransferases (AccD1 to -6) inM. tuberculosisare still not clear, AccD6 in complex with AccA3 can synthesize malonyl-CoA from acetyl-CoA. A series of 10 herbicides that target plant acetyl-CoA carboxylases (ACC) were tested for inhibition of AccD6 and for whole-cell activity againstM. tuberculosis. From the tested herbicides, haloxyfop, an arylophenoxypropionate, showedin vitroinhibition ofM. tuberculosisAccD6, with a 50% inhibitory concentration (IC50) of 21.4 ± 1 μM. Here, we report the crystal structures ofM. tuberculosisAccD6 in the apo form (3.0 Å) and in complex with haloxyfop-R(2.3 Å). The structure ofM. tuberculosisAccD6 in complex with haloxyfop-Rshows two molecules of the inhibitor bound on each AccD6 subunit. These results indicate the potential for developing novel therapeutics for tuberculosis based on herbicides with low human toxicity.

scholarly journals Differences between manganese and magnesium ions with regard to fatty acid biosynthesis, acetyl-coenzyme A carboxylase activity and malonyl-coenzyme A decarboxylation

1970 ◽  
Vol 118 (3) ◽  
pp. 391-399 ◽  
Author(s):  
R. M. Scorpio ◽  
E. J. Masoro
Keyword(s):  
Fatty Acid ◽  
Coenzyme A ◽  
Acetyl Coa Carboxylase ◽  
Biosynthetic Activity ◽  
Acetyl Coa ◽  
Carboxylase Activity ◽  
Bivalent Cation ◽  
Biochemical Basis ◽  
Liver Cytosol ◽  
Malonyl Coa

Fatty acid-biosynthetic activity in rat liver cytosol fractions is much greater when the bivalent cation in the assay system is Mn2+ than when it is Mg2+. This difference between bivalent cations can be abolished if the cytosol fractions are preincubated with isocitrate and the bivalent cation for 30min before assay of fatty acid-biosynthetic activity. In a search for the biochemical basis of this phenomenon, the following differences between Mg2+ and Mn2+ were established: (1) Mn2+ promotes acetyl-CoA carboxylase activity of the protomeric form of the enzyme under conditions in which Mg2+ does not; (2) Mn2++ATP have little inhibitory effect on the polymerization of acetyl-CoA carboxylase whereas Mg2++ATP are markedly inhibitory; (3) under conditions in which utilization of malonyl-CoA in condensation reactions is prevented, the steady-state concentration of malonyl-CoA formed by a cytosol fraction is much greater with Mn2+ than with Mg2+. The role that each of these specific differences between Mn2+ and Mg2+ might play in causing liver cytosol preparations to have greater fatty acid-biosynthetic activity in the presence of Mn2+ is discussed.

scholarly journals Fatty acid ethyl esters in meconium and substance use in adolescence

2021 ◽  
Vol 83 ◽  
pp. 106946
Author(s):  
Meeyoung O. Min ◽  
Sonia Minnes ◽  
Hasina Momotaz ◽  
Lynn T. Singer ◽  
Anna Wasden ◽  
...  
Keyword(s):  
Substance Use ◽  
Fatty Acid ◽  
Fatty Acid Ethyl Esters ◽  
Ethyl Esters

scholarly journals Development and validation of a method for the simultaneous analysis of fatty acid ethyl esters, ethyl sulfate and ethyl glucuronide in neonatal meconium: application in two cases of alcohol consumption during pregnancy

2021 ◽  
Vol 413 (11) ◽  
pp. 3093-3105
Author(s):  
Mateusz Kacper Woźniak ◽  
Laura Banaszkiewicz ◽  
Justyna Aszyk ◽  
Marek Wiergowski ◽  
Iwona Jańczewska ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Fatty Acid ◽  
Alcohol Consumption ◽  
Solid Phase ◽  
Fatty Acid Ethyl Esters ◽  
Ethyl Esters ◽  
Ethyl Glucuronide ◽  
Published Data ◽  
Ethyl Sulfate ◽  
Analytical Approaches

AbstractAlcohol consumption during pregnancy constitutes one of the leading preventable causes of birth defects and neurodevelopmental disorders in the exposed children. Fatty acid ethyl esters (FAEEs), ethyl glucuronide (EtG) and ethyl sulfate (EtS) have been studied as potential biomarkers of alcohol consumption. However, most analytical approaches proposed for their analysis in meconium samples consist of separated extraction procedures requiring the use of two meconium aliquots, which is costly in terms of both time and materials. Therefore, the aim of this study was to develop and validate a method for the simultaneous extraction of 9 FAEEs, EtG and EtS from one meconium aliquot. The sample was homogenized using methanol, and then FAEEs were extracted with hexane while EtG and EtS were isolated using acetonitrile. Then, extracts were applied to solid-phase extraction columns and analysed by gas chromatography mass spectrometry (FAEEs) and liquid chromatography tandem mass spectrometry (EtG and EtS). Calibration curves were linear with r values greater than 0.99. The LODs ranged from 0.8 to 7.5 ng/g for FAEEs and were 0.2 ng/g and 0.8 ng/g for EtS and EtG, respectively. LOQs ranged from 5 to 25 ng/g for FAEEs and were 1 ng/g and 2.5 ng/g for EtS and EtG, respectively. Accuracies and precisions were between 93.8 and 107% and between 3.5 and 9.7%, respectively. The recovery values ranged from 89.1 to 109%. The method proved to be sensitive, specific, simple and fast and allowed for the reduction of the amount of organic solvent used for extraction compared to other published data while higher recoveries were obtained. The method was used for analysis of meconium samples in two cases of mothers who were consuming alcohol during pregnancy.

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