Molecular detection of alpha-glucosidase inhibitor-producing actinomycetes.
In this study, we demonstrate the use of a PCR-based method for the detection of the specific genes involved in natural-product biosynthesis. This method was applied, using specifically designed PCR primers, to the amplification of a gene segment encoding for sedo-heptulose 7-phosphate cyclase, which appears to be involved in the biosynthetic pathways of C7N aminoacyclitol or its keto analogue-containing metabolites, in a variety of actinomycetes species. The sequences of DNA fragments (about 540 bp) obtained from three out of 39 actinomycete strains exhibited a high degree of homology with the sedo-heptulose 7-phosphate cyclase gene, which has been implicated in acarbose biosynthesis. The selective cultivation conditions of this experiment induced the expression of these loci, indicating that the range of C7 aminoacyclitol or its keto analogue-group natural products might be far greater than was previously imagined. Considering that a total of approximately 20 C7 aminoacyclitol metabolites, or its keto analogue-containing metabolites, have been described to date, it appears likely that some of the unknown loci described herein might constitute new classes of C7 aminoacyclitol, or of its keto analogue-containing metabolites. As these metabolites, some of which contain valienamine, are among the most potent antidiabetic agents thus far discovered, the molecular detection of specific metabolite-producing actinomycetes may prove a crucial step in current attempts to expand the scope and diversity of natural-product discovery.
Laboratory of Biotech and Drug Discovery, Chem Tech Research Incorporation (C-TRI), Kowoon Institute of Technology Innovation, Whasung-City 445-743, Republic of Korea.
Chang-Gu
CG
Hyun
Seung-Young
SY
Kim
Jin-Haeng
JH
Hur
Myung-Ji
MJ
Seo
Joo-Won
JW
Suh
Soon-Ok
SO
Kim
D039903:Actinobacteria / Q000235:genetics* / Q000378:metabolism*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism*; D065089:Glycoside Hydrolase Inhibitors; D019746:Intramolecular Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
D001426:Bacterial Proteins; D004791:Enzyme Inhibitors; D065089:Glycoside Hydrolase Inhibitors; D019746:Intramolecular Oxidoreductases; C118743:ACBC protein, Actinoplanes