Carboxyl CoA ligases(CCLs) is an important branch of adenylate synthetase gene family, which mainly has two-step catalytic reactions. Firstly, in the presence of adenosine triphosphate, it can catalyze the pyrophosphorylation of carboxylateswith diffe-rent structures to form corresponding acyl adenosine monophosphate intermediates. Secondly, adenosine monophosphate was replaced by free electrons in the mercaptan group of enzyme A or other acyl receptors by nucleophilic attack to form thioesters. In this study, on the basis of the transcriptome database of Arnebia euchroma, two genes were selected, named AeCCL5(XP_019237476.1) and AeCCL7(XP_019237476.1). Bioinformatics analysis showed that their relative molecular weights were 60.569 kDa and 60.928 kDa, theoretical PI were 8.59 and 8.92, respectively. They both have transmembrane domains but without signal peptide. By multiple sequence alignment and phylogenetic tree analysis, we found that the similarity between AeCCLs and other plant homologous proteins was not high, and the substrate binding sites of AeCCLs were not highly conserved. The reasons might be that the sequence and structure need to adapt to the changes of new substrates in the process of evolution. In this study, the full-length of AeCCL5 and AecCCL7 were cloned into the expression vector pCDFDuet-1. The proteins of AeCCL5 and AeCCL7 with His-tag were expressed in Escherichia coli. The proteins of AeCCL5 and AeCCL7 were purified by nickel column. In vitro enzymatic reactions proved that both AeCCL5 and AeCCL7 can participate in the upstream phenylpropane pathway of shikonin biosynthesisby catalyzing 4-coumaric acid to produce 4-coumarin-CoA, and then to synthesis p-hydroxybenzoic acid, which is an important precursor of shikonin biosynthesis in A. euchroma.
Boraginaceae/genetics* ; Cloning, Molecular ; Coenzyme A ; Coenzyme A Ligases/genetics* ; Ligases ; Phylogeny

No abstract available.
Multiple Acyl Coenzyme A Dehydrogenase Deficiency*

Lovastatin is a potent inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, which catalyzes the conversion of hydroxymethylglutaryl-coenzyme A to mevalonate, anearly and rate-limiting step in the synthesis of cholesterol. We studied the therapeutic effect and safety of lovastatin in 18 patients with nonfamilial primary hypercholesterolemia. Patients received 20mg/day lovastatin therapy as a single evening dose. If the total cholesterol level exceeded 200mg/dl after 2weeks of lovastatin therapy, the dosage of lovastatin was doubled. Mean percent total cholesterol level reductions from baseline were 26.4% and 31.9% after 4, and 8 weeks of lovastatin therapy respectively. Mean percent HDL-cholesterol level increase from baseline were 12% and 13% after 4, and 8 weeks of lovastatin therapy respectively. Adverse effects attributable to lovastatin were mild and temporary and no patient was withdrawn from therapy. We concluded that lovastatin was a well tolerated and effective agent for the treatment of nonfamilial primary hypercholesterolemia. Further studies are needed to establish the long-term safety and effectiveness of this drug.
Cholesterol ; Coenzyme A ; Humans ; Hypercholesterolemia* ; Lovastatin ; Mevalonic Acid ; Oxidoreductases

A new hypolipidemic drug, pravastatin, hydroxymethylglutaryl coenzyme A reductase inhibitor was administered to 33 patients with primary hyperlipidemia, 10mg daily for 8 weeks and sequential changes of lipid profile were analysed as follows. 1) Mean value at baseline period of total cholesterol, triglyceride, high and low density lipoprotein cholesterol were 260, 220, 51 and 163mg/dl respectively. 2) Total cholesterol showed 21% decrease at the end of 8 weeks and that of LDL-cholesterol were 30%. 3) Triglyceride decreased 16% at the end of 8 weeks and increment of HDL-cholesterol was 8% at the end of 8 weeks. 4) No serious side reactions were observed except one patient, who showed generalized skin rash which last 3 days and did not prevent further medication. In conclusion, pravastatin is a safe and useful hypolipidemic agent for the patient with primary hyperlipidemia.
Cholesterol ; Cholesterol, LDL ; Coenzyme A ; Exanthema ; Humans ; Hyperlipidemias* ; Oxidoreductases ; Pravastatin* ; Triglycerides

We report the biochemical profiling in two siblings with mitochondrial 2-methylacetoacetyl-CoA thiolase deficiency. Organic aciduria typical of this rare inborn error metabolism was found when the elder sibling presented with an episode of severe ketoacidosis at 20 months of age, which consisted of excessive excretion of ketones, tiglylglycine, 2-methyl-3-hydroxybutyrate, and 2-methylacetoacetate. Blood acylcarnitiness profile showed elevation of C5OH-carnitine, which represents 2-methyl-3-hydroxybutyrylcarnitine. A similar biochemical profile was identified in the younger sibling during screening although he had only mild clinical symptoms. Both patients reported a favourable outcome on follow-up.
2-methylacetoacetyl-coenzyme A ; deficiency ; Acetyl-CoA C-Acetyltransferase ; Biochemical ; Siblings

Mitochondrial 3-hydroxy-3-methylglutaryl CoA synthase deficiency (HMCSD) is caused by HMGCS2 gene mutation. This paper reports the clinical and genetic features of an infant with this disease. The 8-month-old female infant was admitted to the hospital with diarrhea for 1 week and fever and convulsion for 1 day. The child presented with seizures, acidosis, hypoglycemia, abnormal liver function, myocardial injury and coagulation dysfunction. The new homozygous mutation c.1502G>A(p.R501Q) in the HMGCS2 gene was found in the infant by genetic testing. The mutant gene was found to be harmful by bioinformatics software analysis. Urine organic acid analysis indicated that 4-hydroxy-6-methyl-2-pyranone was significantly increased, which was consistent with the results of genetic testing. The infant was definitely diagnosed with HMCSD.
Acyl Coenzyme A ; Female ; Humans ; Hypoglycemia ; Infant ; Mitochondria ; Mutation

Child ; Humans ; Multiple Acyl Coenzyme A Dehydrogenase Deficiency/genetics* ; Mutation

Raspberry ketones have important therapeutic properties such as anti-influenza and prevention of diabetes. In order to obtain raspberry ketone from Chlamydomonas reinhardtii, two enzymes catalyzing the last two steps of raspberry ketone synthesis, i.e. 4-coumaryl-CoA ligase (4CL) and polyketide synthase (PKS1), were fused using a glycine-serine-glycine (GSG) tripeptide linker to construct an expression vector pChla-4CL-PKS1. The fusion gene 4CL-PKS1 driven by a PSAD promoter was transformed into a wild-type (CC125) and a cell wall-deficient C. reinhardtii (CC425) by electroporation. The results showed the recombinant C. reinhardtii strain CC125 and CC425 with 4CL-PKS1 produced raspberry ketone at a level of 6.7 μg/g (fresh weight) and 5.9 μg/g (fresh weight), respectively, both were higher than that of the native raspberry ketone producing plants (2-4 μg/g).
Acyl Coenzyme A ; Butanones ; Chlamydomonas reinhardtii/genetics* ; Ligases ; Polyketide Synthases

The pyruvate dehydrogenase complex (PDC) is an emerging target for the treatment of metabolic syndrome. To maintain a steady-state concentration of adenosine triphosphate during the feed-fast cycle, cells require efficient utilization of fatty acid and glucose, which is controlled by the PDC. The PDC converts pyruvate, coenzyme A (CoA), and oxidized nicotinamide adenine dinucleotide (NAD+) into acetyl-CoA, reduced form of nicotinamide adenine dinucleotide (NADH), and carbon dioxide. The activity of the PDC is up- and down-regulated by pyruvate dehydrogenase kinase and pyruvate dehydrogenase phosphatase, respectively. In addition, pyruvate is a key intermediate of glucose oxidation and an important precursor for the synthesis of glucose, glycerol, fatty acids, and nonessential amino acids.
Acetyl Coenzyme A ; Adenosine Triphosphate ; Amino Acids ; Carbon Dioxide ; Coenzyme A ; Diabetes Mellitus ; Fatty Acids ; Glucose ; Glycerol ; NAD ; Obesity* ; Oxidoreductases* ; Phosphotransferases* ; Pyruvate Dehydrogenase (Lipoamide)-Phosphatase ; Pyruvate Dehydrogenase Complex ; Pyruvic Acid*

BACKGROUND: N-acetyl transferase (NAT) inactivates the pro-drug isoniazid (INH) to N-acetyl INH through a process of acetylation, and confers low-level resistance to INH in Mycobacterium tuberculosis (MTB). Similar to NAT of MTB, NAT2 in humans performs the same function of acetylation. Rapid acetylators, may not respond to INH treatment efficiently, and could be a potential risk factor, for the development of INH resistance in humans. METHODS: To understand the contribution of NAT of MTB and NAT2 of humans in developing INH resistance using in silico approaches, in this study, the wild type (WT) and mutant (MT)-NATs of MTB, and humans, were modeled and docked, with substrates and product (acetyl CoA, INH, and acetyl INH). The MT models were built, using templates 4BGF of MTB, and 2PFR of humans. RESULTS: On the basis of docking results of MTB-NAT, it can be suggested that in comparison to the WT, binding affinity of MT-G207R, was found to be lower with acetyl CoA, and higher with acetyl-INH and INH. In case of MT-NAT2 from humans, the pattern of score with respect to acetyl CoA and acetyl-INH, was similar to MT-NAT of MTB, but revealed a decrease in INH score. CONCLUSION: In MTB, MT-NAT revealed high affinity towards acetyl-INH, which can be interpreted as increased formation of acetyl-INH, and therefore, may lead to INH resistance through inactivation of INH. Similarly, in MT-NAT2 (rapid acetylators), acetylation occurs rapidly, serving as a possible risk factor for developing INH resistance in humans.
Acetyl Coenzyme A ; Acetylation ; Computer Simulation* ; Humans* ; Isoniazid* ; Mycobacterium tuberculosis* ; Mycobacterium* ; Risk Factors ; Transferases*