| Budget Amount *help |
¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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| Outline of Final Research Achievements |
In this study, we aimed to elucidate the physiological functions of bacterial glycolipids by functional analysis of ceramide glucuronosyltransferase (Cer GlcAT) of Zymomonas mobilis and diacylglycerol glucuronosyltransferase (DAG GlcAT) of Pseudomonas aeruginosa.
We first cloned and expressed Z. mobilis Cer GlcAT in E. coli and purified the enzyme using Ni-affinity and gel filtration chromatography. Using the purified enzyme, we revealed that the enzyme was highly specific for UDP-glucuronic acid and ceramide as donor and acceptor substrates, respectively. We also established the knockout strain and elucidated that this enzyme was the sole Cer GlcAT of Z. mobilis. Next, we cloned PA0842 of P. aeruginonsa and expressed it in E. coli. We detected the DAG GlcAT activity when UDP-glucuronic acid and diacylglycerol were used as donor and acceptor substrates, respectively. Furthermore, we found that PA0842 was induced to produce glucuronosyl DAG under phosphate-deficient conditions.
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