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Molecular characterization of the acyl-coenzyme A:isopenicillin N acyltransferase gene (penDE) from Penicillium chrysogenum and Aspergillus nidulans and activity of recombinant enzyme in Escherichia coli.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Tobin, Matthew B. Fleming, Mark D. Skatrud, Paul Luther Miller, J. Richard |
| Copyright Year | 1990 |
| Abstract | The final step in the biosynthesis of beta-lactam antibiotics in Penicillium chrysogenum and Aspergillus nidulans involves removal of the L-alpha-aminoadipyl side chain from isopenicillin N (IPN) and exchange with a nonpolar side chain. The enzyme catalyzing this reaction, acyl-coenzyme A:isopenicillin N acyltransferase (acyltransferase), was purified from P. chrysogenum and A. nidulans. Based on NH2-terminal amino acid sequence information, the acyltransferase gene (penDE) from P. chrysogenum and A. nidulans were cloned. In both organisms, penDE was located immediately downstream from the isopenicillin N synthetase gene (pcbC) and consisted of four exons encoding an enzyme of 357 amino acids (approximately 40 kilodaltons [kDa]). The DNA coding sequences showed approximately 73% identity, while the amino acid sequences were approximately 76% identical. Noncoding DNA regions (including the region between pcbC and penDE) were not conserved. Acyltransferase activity from Escherichia coli producing the 40-kDa protein accepted either 6-aminopenicillanic acid or IPN as the substrate and made a penicillinase-sensitive antibiotic in the presence of phenylacetyl coenzyme A. Therefore, a single gene is responsible for converting IPN to penicillin G. The active form of the enzyme may result from processing of the 40-kDa monomeric precursor to a heterodimer containing subunits of 11 and 29 kDa. |
| File Format | PDF HTM / HTML |
| DOI | 10.1128/jb.172.10.5908-5914.1990 |
| PubMed reference number | 2120195 |
| Journal | Medline |
| Volume Number | 172 |
| Issue Number | 10 |
| Alternate Webpage(s) | http://jb.asm.org/content/172/10/5908.full.pdf |
| Journal | Journal of bacteriology |
| Language | English |
| Access Restriction | Open |
| Content Type | Text |
| Resource Type | Article |
