Analysis of intracellular short organic acid‐coenzyme A esters from actinomycetes using liquid chromatography‐electrospray ionization‐mass spectrometry

Park, Je Won; Jung, Won Seok; Park, Sung Ryeol; Park, Byoung Chul; Yoon, Yeo Joon

doi:10.1002/jms.1240

Cited by 63 publications

(66 citation statements)

References 26 publications

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“…It was obtained by the loss of a fragment (m/z ϭ 507 Da) from the parent ion. Park et al (33) used the constant decrease of m/z ϭ 507 Da from parent ions of different CoA esters to ascertain the identity of the expected CoA derivative in their study. Therefore, these researchers calculated the expected mass of the second daughter by subtracting m/z ϭ 507 Da from the respective parent ion.…”

Section: Discussionmentioning

confidence: 99%

Novel Reaction of Succinyl Coenzyme A (Succinyl-CoA) Synthetase: Activation of 3-Sulfinopropionate to 3-Sulfinopropionyl-CoA in Advenella mimigardefordensis Strain DPN7 ^T during Degradation of 3,3′-Dithiodipropionic Acid

et al. 2011

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The sucCD gene of Advenella mimigardefordensis strain DPN7 T encodes a succinyl coenzyme A (succinyl-CoA) synthetase homologue (EC 6.2.1.4 or EC 6.2.1.5) that recognizes, in addition to succinate, the structural analogues 3-sulfinopropionate (3SP) and itaconate as substrates. Accumulation of 3SP during 3,3-dithiodipropionic acid (DTDP) degradation was observed in Tn5::mob-induced mutants of A. mimigardefordensis strain DPN7T disrupted in sucCD and in the defined deletion mutant A. mimigardefordensis ⌬sucCD. These mutants were impaired in growth with DTDP and 3SP as the sole carbon source. Hence, it was proposed that the succinyl-CoA synthetase homologue in A. mimigardefordensis strain DPN7T activates 3SP to the corresponding CoA-thioester (3SP-CoA). The putative genes coding for A. mimigardefordensis succinyl-CoA synthetase (SucCD Am ) were cloned and heterologously expressed in Escherichia coli BL21(DE3)/pLysS. Purification and characterization of the enzyme confirmed its involvement during degradation of DTDP. 3SP, the cleavage product of DTDP, was converted into 3SP-CoA by the purified enzyme, as demonstrated by in vitro enzyme assays. The structure of 3SP-CoA was verified by using liquid chromatography-electrospray ionization-mass spectrometry. SucCD Am is Mg 2؉ or Mn 2؉ dependent and unspecific regarding ATP or GTP. In kinetic studies the enzyme showed highest enzyme activity and substrate affinity with succinate (V max ‫؍‬ 9.85 ؎ 0.14 mol min ؊1 mg ؊1 , K m ‫؍‬ 0.143 ؎ 0.001 mM). In comparison to succinate, activity with 3SP was only ca. 1.2% (V max ‫؍‬ 0.12 ؎ 0.01 mol min ؊1 mg ؊1 ) and the affinity was 6-fold lower (K m ‫؍‬ 0.818 ؎ 0.046 mM). Based on the present results, we conclude that SucCD Am is physiologically associated with the citric acid cycle but is mandatory for the catabolic pathway of DTDP and its degradation intermediate 3SP.3,3Ј-Dithiodipropionic acid (DTDP) is an organic disulfide and a precursor for the production of polythioesters (PTEs) by bacteria (25). Further applications for DTDP are thermodynamic studies (40), development of secondary batteries (52), amino acid analysis (53), and the construction of self-assembling monolayers (10). Microbial production of PTEs from simple carbon sources and inorganic sulfur is currently not possible. Knowledge of the catabolism of organic sulfur compounds in bacteria could provide a reasonable strategy to engineer strains suitable for PTE production. A first step in this direction was the isolation of bacteria able to utilize DTDP as the sole source of carbon and energy. Advenella mimigardefordensis strain DPN7T , a betaproteobacterium, found in mature compost in a waste management facility was one of the isolates (15,56

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Section: Discussionmentioning

confidence: 99%

Novel Reaction of Succinyl Coenzyme A (Succinyl-CoA) Synthetase: Activation of 3-Sulfinopropionate to 3-Sulfinopropionyl-CoA in Advenella mimigardefordensis Strain DPN7 ^T during Degradation of 3,3′-Dithiodipropionic Acid

et al. 2011

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“…However, most methods only assay either short-, medium-, or long-chain acyl-CoAs (9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19). The best approach for acyl-CoA quantitation, so far, is to measure the target acyl-CoAs from short to long carbon chain length in a single method ( 20,21 ).…”

Section: Lc-ms/ms Is the Most Sensitive Methods To Analyze Acylcoas Anmentioning

confidence: 99%

Novel approach in LC-MS/MS using MRM to generate a full profile of acyl-CoAs: discovery of acyl-dephospho-CoAs

Zhang

Berthiaume

et al. 2014

Journal of Lipid Research

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This article is available online at http://www.jlr.org Acyl-CoAs are a class of important molecules that play essential roles in many physiological processes ( 1 ), such as fatty acid oxidation, lipid synthesis/remodeling, ketone body synthesis, xenobiotic metabolism, and signaling pathways. Classically, acyl-CoAs such as free CoA, acetyl-CoA, and malonyl-CoA are recognized as regulators of metabolic fl ux. The ratio of acetyl-CoA versus free CoA tightly regulates glycolysis and fatty acid oxidation. Malonyl-CoA attenuates fatty acid oxidation by inhibiting acyl-CoA transport into the mitochondrion for oxidation and is utilized in fatty acid synthesis when its concentration is elevated ( 2 ). Many proteins and genes are dynamically regulated by deacylation and acylation via various acyl-CoAs, such as acetyl-CoA, succinyl-CoA, palmitoyl-CoA, etc. ( 3 ). However, acyl-CoAs present in the cell are diverse and may involve molecules beyond fatty acids and their oxidized derivatives. The metabolism of xenobiotics can also lead to the formation of acyl-CoAs, as demonstrated in our previous work on the metabolism of 4-hydroxy acids from C 4 to C 11 in perfused rat liver. The identifi cation of these novel acyl-CoAs extends our understanding of the new catabolic pathways involved in the disposal of 4-hydroxy acids including drugs of abuse and lipid peroxidation products ( 4-8 ).Acyl-CoAs are exclusive to the intracellular metabolites, and the profi le of these biomolecules is indicative of the local metabolic status. Each organ has its specifi c physiological roles with different energetic demands, and therefore, would be expected to exhibit a unique acyl-CoA profi le. The heart preferentially utilizes fatty acids to meet the ATP demand of mechanical contraction. Glucose and/or ketone bodies serve as the primary substrate of the brain for energy

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“…The SPE method for acyl-CoAs was according to the method described by Park et al [16] with OASIS HLB 3 cc/60 mg SPE column.…”

Section: Solid-phase Extraction (Spe) Methods For Acyl-coasmentioning

confidence: 99%

13C-assisted metabolomics analysis reveals the positive correlation between specific erythromycin production rate and intracellular propionyl-CoA pool size in Saccharopolyspora erythraea

Hong

Mou

Liu

et al. 2017

Bioprocess Biosyst Eng

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Metabolomics analysis is extremely essential to explore the metabolism characteristics of Saccharopolyspora erythraea. The lack of suitable methods for the determination of intracellular metabolites, however, hinders the application of metabolomics analysis for S. erythraea. Acyl-CoAs are important precursors of erythromycin; phosphorylated sugars are intermediate metabolites in EMP pathway or PPP pathway; organic acids are intermediate metabolites in TCA cycle. Reliable determination methods for intracellular acyl-CoAs, phosphorylated sugars, and organic acids of S. erythraea were designed and validated in this study. Using the optimized determination methods, the pool sizes of intracellular metabolites during an erythromycin fermentation process were precisely quantified by isotope dilution mass spectroscopy method. The quantification results showed that the specific erythromycin production rate was positively correlated with the pool sizes of propionyl-CoA as well as many other intracellular metabolites. The experiment under the condition without propanol, which is a precursor of propionyl-CoA and an important substrate in industrial erythromycin production process, also corroborated the correlation between specific erythromycin production rate and intracellular propionyl-CoA pool size. As far as we know, this is the first paper to conduct the metabolomics analysis of S. erythraea, which makes the metabolomics analysis of S. erythraea in the industrial erythromycin production process possible.

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Analysis of intracellular short organic acid‐coenzyme A esters from actinomycetes using liquid chromatography‐electrospray ionization‐mass spectrometry

Cited by 63 publications

References 26 publications

Novel Reaction of Succinyl Coenzyme A (Succinyl-CoA) Synthetase: Activation of 3-Sulfinopropionate to 3-Sulfinopropionyl-CoA in Advenella mimigardefordensis Strain DPN7 ^T during Degradation of 3,3′-Dithiodipropionic Acid

Novel Reaction of Succinyl Coenzyme A (Succinyl-CoA) Synthetase: Activation of 3-Sulfinopropionate to 3-Sulfinopropionyl-CoA in Advenella mimigardefordensis Strain DPN7 ^T during Degradation of 3,3′-Dithiodipropionic Acid

Novel approach in LC-MS/MS using MRM to generate a full profile of acyl-CoAs: discovery of acyl-dephospho-CoAs

13C-assisted metabolomics analysis reveals the positive correlation between specific erythromycin production rate and intracellular propionyl-CoA pool size in Saccharopolyspora erythraea

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Analysis of intracellular short organic acid‐coenzyme A esters from actinomycetes using liquid chromatography‐electrospray ionization‐mass spectrometry

Cited by 63 publications

References 26 publications

Novel Reaction of Succinyl Coenzyme A (Succinyl-CoA) Synthetase: Activation of 3-Sulfinopropionate to 3-Sulfinopropionyl-CoA in Advenella mimigardefordensis Strain DPN7 T during Degradation of 3,3′-Dithiodipropionic Acid

Novel Reaction of Succinyl Coenzyme A (Succinyl-CoA) Synthetase: Activation of 3-Sulfinopropionate to 3-Sulfinopropionyl-CoA in Advenella mimigardefordensis Strain DPN7 T during Degradation of 3,3′-Dithiodipropionic Acid

Novel approach in LC-MS/MS using MRM to generate a full profile of acyl-CoAs: discovery of acyl-dephospho-CoAs

13C-assisted metabolomics analysis reveals the positive correlation between specific erythromycin production rate and intracellular propionyl-CoA pool size in Saccharopolyspora erythraea

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Novel Reaction of Succinyl Coenzyme A (Succinyl-CoA) Synthetase: Activation of 3-Sulfinopropionate to 3-Sulfinopropionyl-CoA in Advenella mimigardefordensis Strain DPN7 ^T during Degradation of 3,3′-Dithiodipropionic Acid

Novel Reaction of Succinyl Coenzyme A (Succinyl-CoA) Synthetase: Activation of 3-Sulfinopropionate to 3-Sulfinopropionyl-CoA in Advenella mimigardefordensis Strain DPN7 ^T during Degradation of 3,3′-Dithiodipropionic Acid