Biochemical characterization of the Nocardia lactamdurans ACV synthetase
| العنوان: | Biochemical characterization of the Nocardia lactamdurans ACV synthetase |
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| المؤلفون: | Riccardo Iacovelli, Roel A. L. Bovenberg, Arnold J. M. Driessen, Reto D. Zwahlen |
| المساهمون: | Molecular Microbiology, Groningen Biomolecular Sciences and Biotechnology |
| المصدر: | PLoS ONE PLoS ONE, Vol 15, Iss 4, p e0231290 (2020) PLoS ONE, 15(4):e0231290. PUBLIC LIBRARY SCIENCE |
| بيانات النشر: | Public Library of Science (PLoS), 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | 0301 basic medicine, Enzyme Purification, Protein Sequencing, Tripeptide, Pathology and Laboratory Medicine, Protein Engineering, Biochemistry, 01 natural sciences, Nocardia, Substrate Specificity, CONDENSATION, Database and Informatics Methods, Medicine and Health Sciences, PEPTIDE SYNTHETASES, Peptide bond, Peptide Synthases, Peptide sequence, chemistry.chemical_classification, Multidisciplinary, biology, CEPHALOSPORIUM-ACREMONIUM, Bacterial Pathogens, Enzymes, Medical Microbiology, Amino Acid Analysis, Medicine, Engineering and Technology, Pathogens, Sequence Analysis, ANTIBIOTICS, Research Article, DOMAINS, ENZYME, Bioinformatics, Science, ACV synthetase, Protein domain, Research and Analysis Methods, Microbiology, 03 medical and health sciences, Protein Domains, Sequence Motif Analysis, Nonribosomal peptide, DELTA-(L-ALPHA-AMINOADIPYL)-L-CYSTEINYL-D-VALINE SYNTHETASE, Amino Acid Sequence, Molecular Biology Techniques, Sequencing Techniques, Microbial Pathogens, Molecular Biology, Molecular Biology Assays and Analysis Techniques, PURIFICATION, Bacteria, 010405 organic chemistry, Organisms, Biology and Life Sciences, Proteins, Protein engineering, 0104 chemical sciences, 030104 developmental biology, chemistry, PENICILLIN BIOSYNTHESIS, Enzymology, biology.protein, Heterologous expression, Peptides, Sequence Alignment, CHRYSOGENUM, Purification Techniques |
| الوصف: | The L-δ-(α-aminoadipoyl)-L-cysteinyl-D-valine synthetase (ACVS) is a nonribosomal peptide synthetase (NRPS) that fulfills a crucial role in the synthesis of β-lactams. Although some of the enzymological aspects of this enzyme have been elucidated, its large size, at over 400 kDa, has hampered heterologous expression and stable purification attempts. Here we have successfully overexpressed the Nocardia lactamdurans ACVS in E. coli HM0079. The protein was purified to homogeneity and characterized for tripeptide formation with a focus on the substrate specificity of the three modules. The first L-α-aminoadipic acid-activating module is highly specific, whereas the modules for L-cysteine and L-valine are more promiscuous. Engineering of the first module of ACVS confirmed the strict specificity observed towards its substrate, which can be understood in terms of the non-canonical peptide bond position. |
| وصف الملف: | application/pdf |
| تدمد: | 1932-6203 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::2b59d41a10b1ab624d9ceeb75c48dadf https://doi.org/10.1371/journal.pone.0231290 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....2b59d41a10b1ab624d9ceeb75c48dadf |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 19326203 |
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