Post-PKS tailoring steps of a disaccharide-containing polyene NPP in Pseudonocardia autotrophica.
A novel polyene compound NPP identified in a rare actinomycetes, Pseudonocardia autotrophica KCTC9441, was shown to contain an aglycone identical to nystatin but to harbor a unique di-sugar moiety, mycosaminyl-(α1-4)-N-acetyl-glucosamine, which led to higher solubility and reduced hemolytic activity...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2015-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0123270 |
| _version_ | 1819147225161269248 |
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| author | Hye-Jin Kim Min-Kyung Kim Mi-Jin Lee Hyung-Jin Won Si-Sun Choi Eung-Soo Kim |
| author_facet | Hye-Jin Kim Min-Kyung Kim Mi-Jin Lee Hyung-Jin Won Si-Sun Choi Eung-Soo Kim |
| author_sort | Hye-Jin Kim |
| collection | DOAJ |
| description | A novel polyene compound NPP identified in a rare actinomycetes, Pseudonocardia autotrophica KCTC9441, was shown to contain an aglycone identical to nystatin but to harbor a unique di-sugar moiety, mycosaminyl-(α1-4)-N-acetyl-glucosamine, which led to higher solubility and reduced hemolytic activity. Although the nppDI was proved to be responsible for the transfer of first polyene sugar, mycosamine in NPP biosynthesis, the gene responsible for the second sugar extending glycosyltransferase (GT) as well as NPP post-PKS tailoring mechanism remained unknown. Here, we identified a NPP-specific second sugar extending GT gene named nppY, located at the edge of the NPP biosynthetic gene cluster. Targeted nppY gene deletion and its complementation proved that nppY is indeed responsible for the transfer of second sugar, N-acetyl-glucosamine in NPP biosynthesis. Site-directed mutagenesis on nppY also revealed several amino acid residues critical for NppY GT function. Moreover, a combination of deletions and complementations of two GT genes (nppDI and nppY) and one P450 hydroxylase gene (nppL) involved in the NPP post-PKS biosynthesis revealed that NPP aglycone is sequentially modified by the two different GTs encoded by nppDI and nppY, respectively, followed by the nppL-driven regio-specific hydroxylation at the NPP C10 position. These results set the stage for the biotechnological application of sugar diversification for the biosynthesis of novel polyene compounds in actinomycetes. |
| first_indexed | 2024-12-22T13:26:26Z |
| format | Article |
| id | doaj.art-a5318c3decc24d1f82b129022e51ee13 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-22T13:26:26Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-a5318c3decc24d1f82b129022e51ee132022-12-21T18:24:18ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-01104e012327010.1371/journal.pone.0123270Post-PKS tailoring steps of a disaccharide-containing polyene NPP in Pseudonocardia autotrophica.Hye-Jin KimMin-Kyung KimMi-Jin LeeHyung-Jin WonSi-Sun ChoiEung-Soo KimA novel polyene compound NPP identified in a rare actinomycetes, Pseudonocardia autotrophica KCTC9441, was shown to contain an aglycone identical to nystatin but to harbor a unique di-sugar moiety, mycosaminyl-(α1-4)-N-acetyl-glucosamine, which led to higher solubility and reduced hemolytic activity. Although the nppDI was proved to be responsible for the transfer of first polyene sugar, mycosamine in NPP biosynthesis, the gene responsible for the second sugar extending glycosyltransferase (GT) as well as NPP post-PKS tailoring mechanism remained unknown. Here, we identified a NPP-specific second sugar extending GT gene named nppY, located at the edge of the NPP biosynthetic gene cluster. Targeted nppY gene deletion and its complementation proved that nppY is indeed responsible for the transfer of second sugar, N-acetyl-glucosamine in NPP biosynthesis. Site-directed mutagenesis on nppY also revealed several amino acid residues critical for NppY GT function. Moreover, a combination of deletions and complementations of two GT genes (nppDI and nppY) and one P450 hydroxylase gene (nppL) involved in the NPP post-PKS biosynthesis revealed that NPP aglycone is sequentially modified by the two different GTs encoded by nppDI and nppY, respectively, followed by the nppL-driven regio-specific hydroxylation at the NPP C10 position. These results set the stage for the biotechnological application of sugar diversification for the biosynthesis of novel polyene compounds in actinomycetes.https://doi.org/10.1371/journal.pone.0123270 |
| spellingShingle | Hye-Jin Kim Min-Kyung Kim Mi-Jin Lee Hyung-Jin Won Si-Sun Choi Eung-Soo Kim Post-PKS tailoring steps of a disaccharide-containing polyene NPP in Pseudonocardia autotrophica. PLoS ONE |
| title | Post-PKS tailoring steps of a disaccharide-containing polyene NPP in Pseudonocardia autotrophica. |
| title_full | Post-PKS tailoring steps of a disaccharide-containing polyene NPP in Pseudonocardia autotrophica. |
| title_fullStr | Post-PKS tailoring steps of a disaccharide-containing polyene NPP in Pseudonocardia autotrophica. |
| title_full_unstemmed | Post-PKS tailoring steps of a disaccharide-containing polyene NPP in Pseudonocardia autotrophica. |
| title_short | Post-PKS tailoring steps of a disaccharide-containing polyene NPP in Pseudonocardia autotrophica. |
| title_sort | post pks tailoring steps of a disaccharide containing polyene npp in pseudonocardia autotrophica |
| url | https://doi.org/10.1371/journal.pone.0123270 |
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