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Enhanced PHB Accumulation in Photosystem- and Respiration-defective Mutants of a Cyanobacterium Synechocystis sp. PCC 6803  

Kim Soo-Youn (Department of Microbiology, Chungnam National University)
Choi Gang Guk (Department of Microbiology, Chungnam National University)
Park Youn Il (Department of Biology, Chungnam National University)
Park Young Mok (Biomolecular Research Team, Korea Basic Science Institute)
Yang Young Ki (Department of Genetic Engineering, Chosun University)
Rhee Young Ha (Department of Microbiology, Chungnam National University)
Publication Information
Korean Journal of Microbiology / v.41, no.1, 2005 , pp. 67-73 More about this Journal
Abstract
Photoautotrophic bacteria are promising candidates for the production of poly(3-hydroxybutyrate) (PHB) since they can address the critical problem of substrate costs. In this study, we isolated 25 Tn5-inserted mutants of the Synechocystis sp. PCC 6803 which showed enhanced PHB accumulation compared to the wild-type strain. After 5-days cultivation under nitrogen-limited mixotrophic conditions, the intracellular levels of PHB content in these mutants reached up to $10-30\%$ of dry cell weight (DCW) comparable to $4\%$ of DCW in the wild-type strain. Using the method of inverse PCR, the affected genes of the mutants were mapped on the completely known genome sequence of Synechocystis sp. PCC 6803. As a result, the increased PHB accumulation in 5 mutants were found to be resulted from defects of genes coding for NADH-ubiquinone oxidoreductase, O-succinylbenzoic-CoA ligase, photosystem II PsbT protein or histidine kinase, which are involved in photosystem in thylakoid inner membrane of the cell. The values of $NAD(P)H/NAD(P)^+$ ratio in the cells of these mutants were much higher than that of the wild-type strain as measured by using pulse-amplitude modulated fluorometer, suggesting that PHB synthesis could be enhanced by increasing the level of cellular NAD(P)H which is a limiting substrate for NADPH-dependent acetoacetyl-CoA reductase. From these results, it is likely that NAD(P)H would be a limiting factor for PHB synthesis in Synechocystis sp. PCC 6803.
Keywords
NADPH/NADP+ ratio; poly(3-hydroxybutyrate); Synechocystis sp. PCC 6803; transposon mutagenesis;
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