KMID : 1094720080130060738
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Biotechnology and Bioprocess Engineering 2008 Volume.13 No. 6 p.738 ~ p.744
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Expression profiling of Streptomyces peucetius metabolic genes using DNA microarray analysis
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Yang Joon-Ryeol
Song Eun-Jung Kim Byung-Gee Kim Eung-Soo Sohng Jae-Kyung Oh Min-Kyu
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Abstract
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Doxorubicin (DXR) and daunorubicin (DNR) are anthracycline antibiotics produced by Streptomyces peucetius and widely used as cancer chemotherapeutic agents. To improve their productivity, regulation of DXR/DNR synthesis genes as well as central metabolic pathway genes must be understood more clearly. So far, studies have focused on DXR/DNR gene regulation. To investigate the correlation between the central metabolic pathway genes and DXR/DNR productivity, we selected 265 genes involved in glycolysis, fermentation, the citric acid cycle, butanoate metabolism, etc., and searched for their sequences in the S. peucetius genome by comparing gene sequences to those of Streptomyces coelicolor. The homologous genes were amplified by PCR and arrayed on glass microarray slides. Gene expression was monitored under two different growth media conditions, R2YE and NDYE. Genes involved in the production of malonyl-CoA and propionyl-CoA, the main precursors for doxorubicin synthesis, were mainly upregulated in NDYE media. Genes related to acetyl-CoA and the urea cycle were also upregulated. These changes in gene expression were confirmed by real-time RT-PCR.
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KEYWORD
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Streptomyces peucetius, DNA microarray, doxorubicin, metabolic gene
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