Usnea longissima has a long history of use as a traditional medicine. Several bioactive compounds, primarily belonging to the polyketide family, have been isolated from U. longissima. However, the genes for the biosynthesis of these compounds are yet to be identified. In the present study, three dif...
Usnea longissima has a long history of use as a traditional medicine. Several bioactive compounds, primarily belonging to the polyketide family, have been isolated from U. longissima. However, the genes for the biosynthesis of these compounds are yet to be identified. In the present study, three different types of non-reducing polyketide synthases (UlPKS2, UlPKS4, and UlPKS6) were identified from a cultured lichen-forming fungus of U. longissima. Phylogenetic analysis of product template domains showed that UlPKS2 and UlPKS4 belong to group IV, which includes the non-reducing polyketide synthases with an methyltransferase (MeT) domain that are involved in methylorcinol-based compound synthesis; UlPKS6 was found to belong to group I, which includes the non-reducing polyketide synthases that synthesize single aromatic ring polyketides, such as orsellinic acid. Reverse transcriptase-PCR analysis demonstrated that UlPKS2 and UlPKS4 were upregulated by sucrose; UlPKS6 was downregulated by asparagine, glycine, and alanine.
Usnea longissima has a long history of use as a traditional medicine. Several bioactive compounds, primarily belonging to the polyketide family, have been isolated from U. longissima. However, the genes for the biosynthesis of these compounds are yet to be identified. In the present study, three different types of non-reducing polyketide synthases (UlPKS2, UlPKS4, and UlPKS6) were identified from a cultured lichen-forming fungus of U. longissima. Phylogenetic analysis of product template domains showed that UlPKS2 and UlPKS4 belong to group IV, which includes the non-reducing polyketide synthases with an methyltransferase (MeT) domain that are involved in methylorcinol-based compound synthesis; UlPKS6 was found to belong to group I, which includes the non-reducing polyketide synthases that synthesize single aromatic ring polyketides, such as orsellinic acid. Reverse transcriptase-PCR analysis demonstrated that UlPKS2 and UlPKS4 were upregulated by sucrose; UlPKS6 was downregulated by asparagine, glycine, and alanine.
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대상 데이터
The putative function of these ORFs was determined using a basic local alignment search tool (BLAST). The conserved domain was confirmed using the CDD-Search/PRS-BLAST (http://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi).
Of the clones, three contained a partial KS domain sequence (NRks57, LC3551, and MTc14). The NRks57 clone was obtained using degenerate primers NR3KSF and NR3KSR. The closest NRks57 homologue found in the GenBank database was a putative NR-PKS KS domain fragment from Coccidioides posadasii (87% identity; GenBank accession No.
After screening of the genomic library, a clone containing a complete PKS fragment was chosen for further sequencing. A total of three PKSs, UlPKS2, UlPKS4, and UlPKS6, were obtained from the sequence analysis.
이론/모형
com). The putative function of these ORFs was determined using a basic local alignment search tool (BLAST). The conserved domain was confirmed using the CDD-Search/PRS-BLAST (http://www.
2 program. A phylogenetic tree was constructed using the minimum evolution method.
The mixture of symbiotic organisms may provide imprecise information during the genetic analysis of LFF. Therefore, we isolated and cultured the LFF of U. longissima, using the Yamamoto method [28], to acquire genetic information.
성능/효과
Comparison of UlPKS2 with corresponding domains of other fungal PKSs showed conservation of active site residues in the KS (DTACSASAV), AT (GHSFGQ), PT (GHEVVDQTLCPAS), ACP1 (NIDSLLATE) ACP2 (GVDSLLATE), and TE (GWSSGGHL) domains. UlPKS2 was deposited in GenBank with the accession No.
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