Proteome analysis of Halobacterium salinarum and characterization of proteins related to the degradation of isopropyl alcohol.
We reported in a previous study that proteomic approach, coupled with genomic techniques, could be used to screen and develop multiple candidates for halophilic enzymes from Halobacterium salinarum. In order to evaluate the biodegradation of isopropyl alcohol (IPA) by H. salinarum, the amounts of residual IPA and acetone generated in the growth media were determined using a gas chromatography-flame ionization detector (GC-FID). The protein expression profiles of cells which had been cultured with IPA were obtained with the two-dimensional gel electrophoresis. Proteins evidencing different expression levels in the presence of 0.5% IPA were identified by electrospray ionization-quadruple-time of flight (ESI-Q-TOF) mass spectrometry. We found 12 proteins which were down-regulated, and another 12 proteins which were up-regulated, in the presence of 0.5% IPA and we further identified 17 proteins among them using ESI-TOF MS/MS. Among these identified proteins, we selected glyceraldehyde 3-phosphate dehydrogenase (GAPDH) for further characterization as a halophilic enzyme. We have demonstrated for the first time that H. salinarum possesses the ability to degrade IPA and GAPDH was both stable and active at high salt concentrations, with maximum activity occurring at 1 M NaCl, although the optimal salt concentration with regard to the growth of H. salinarum is 4.3 M.
Ha
Dong-Jin
DJ
School of Life Sciences and Biotechnology, Korea University, 1, 5-Ka, Anam-dong, Sungbuk-ku, Seoul 136-701, Korea.
Joo
Won-A
WA
Han
Gi-Yeon
GY
Kim
Chan-Wha
CW
D019840:2-Propanol / Q000378:metabolism* / Q000494:pharmacology; D019843:Archaeal Proteins / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+) / Q000737:chemistry / Q000235:genetics; D019617:Halobacterium salinarum / Q000737:chemistry* / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism*; D020543:Proteome
D019843:Archaeal Proteins; D020543:Proteome; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+); D019840:2-Propanol