https://scholars.lib.ntu.edu.tw/handle/123456789/502738
Title: | Characterization of a Phosphatidylserine Synthase of Vibrio parahaemolyticus | Authors: | Huang, Ru-Yin CHIA-YIN LEE |
Issue Date: | May-2020 | Publisher: | SPRINGER | Journal Volume: | 77 | Journal Issue: | 5 | Start page/Pages: | 710 | Source: | Current microbiology | Abstract: | Phosphatidylserine synthase (Pss) is involved in the metabolic pathway in phospholipid synthesis in different organisms. In this study, Pss expression in Vibrio parahaemolyticus was verified through liquid chromatography tandem-mass spectrometry. To analyze the characteristics of Pss, the recombinant Pss was overexpressed and purified from Escherichia coli. The optimum temperature and pH of Pss were 40 °C and 8, respectively. When reacting with divalent metal, Pss activity decreased. In addition, Pss could not only use cytidine diphosphate diacylglycerol (CDP-DAG, 16:0), but also CDP-DAG (18:1) as a substrate to produce cytidine 5'-monophosphate. Furthermore, the 3D structure of Pss was predicted, and the results revealed that histidine and lysine of the two HKD motifs were present in the catalytic site. Moreover, CDP-DAG (16:0) was docked with the Pss model. To investigate whether the two HKD motifs in Pss are important to its activity, site-directed mutagenesis of histidine was performed. The results revealed that the activities of both H131A and H352A were diminished. Little is known regarding the catalytic site of type I Pss. This is the first report on the biochemical characterization of Pss in V. parahaemolyticus. |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/502738 | ISSN: | 0343-8651 | DOI: | 10.1007/s00284-019-01854-x |
Appears in Collections: | 農業化學系 |
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