Butyryl/Caproyl-CoA:Acetate CoA-transferase: cloning, expression and characterization of the key enzyme involved in medium-chain fatty acid biosynthesis
| العنوان: | Butyryl/Caproyl-CoA:Acetate CoA-transferase: cloning, expression and characterization of the key enzyme involved in medium-chain fatty acid biosynthesis |
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| المؤلفون: | Shengtao Guo, Yong Tao, Qi Lu, Jun Liu, Qinmao Zhou, Decong Zheng, Qingzhuoma Yang |
| المصدر: | Bioscience Reports |
| بيانات النشر: | Portland Press Ltd., 2021. |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | Models, Molecular, Bioinformatics, Protein Conformation, Biophysics, Butyrate, Molecular cloning, Microbiology, Chain elongation, Biochemistry, Substrate Specificity, Active center, Structure-Activity Relationship, chemistry.chemical_compound, Bacterial Proteins, Biosynthesis, Escherichia coli, Cloning, Molecular, Caproates, Molecular Biology, Research Articles, Phylogeny, chemistry.chemical_classification, biology, Medium-chain fatty acids, Fatty acid, Cell Biology, biology.organism_classification, CoA-transferase, Clostridium tyrobutyricum, Enzyme assay, Butyrates, Kinetics, Enzyme, chemistry, Ruminococcaceae bacterium, Mutation, biology.protein, Caproic acid, Acyl Coenzyme A, Coenzyme A-Transferases, Oxidation-Reduction |
| الوصف: | Coenzyme A transferases (CoATs) are important enzymes involved in carbon chain elongation, contributing to medium-chain fatty acid (MCFA) biosynthesis. For example, butyryl-CoA:acetate CoA transferase (BCoAT) is responsible for the final step of butyrate synthesis from butyryl-CoA. However, little is known about caproyl-CoA:acetate CoA-transferase (CCoAT), which is responsible for the final step of caproate synthesis from caproyl-CoA. In the present study, two CoAT genes from Ruminococcaceae bacterium CPB6 and Clostridium tyrobutyricum BEY8 were identified by gene cloning and expression analysis. Enzyme assays and kinetic studies were carried out using butyryl-CoA or caproyl-CoA as the substrate. CPB6-CoAT can catalyze the conversion of both butyryl-CoA into butyrate and caproyl-CoA into caproate, but its catalytic efficiency with caproyl-CoA as the substrate was 3.8-times higher than that with butyryl-CoA. In contrast, BEY8-CoAT had only BCoAT activity, not CCoAT activity. This demonstrated the existence of a specific CCoAT involved in chain elongation via the reverse β-oxidation pathway. Comparative bioinformatics analysis showed the presence of a highly conserved motif (GGQXDFXXGAXX) in CoATs, which is predicted to be the active center. Single point mutations in the conserved motif of CPB6-CoAT (Asp346 and Ala351) led to marked decreases in the activity for butyryl-CoA and caproyl-CoA, indicating that the conserved motif is the active center of CPB6-CoAT and that Asp346 and Ala351 have a significant impact on the enzymatic activity. This work provides insight into the function of CCoAT in caproic acid biosynthesis and improves understanding of the chain elongation pathway for MCFA production. |
| تدمد: | 1573-4935 0144-8463 |
| DOI: | 10.1042/bsr20211135 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::5fbc79b929d617eb68bc1e6ba1e32b28 https://doi.org/10.1042/bsr20211135 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....5fbc79b929d617eb68bc1e6ba1e32b28 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15734935 01448463 |
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| DOI: | 10.1042/bsr20211135 |