Please use this identifier to cite or link to this item: http://scholarbank.nus.edu.sg/handle/10635/13768
Title: Proteome analysis of gentisate-induced responses in Pseudomonas alcaligenes NCIB 9867
Authors: ZHAO BING
Keywords: Gentisate pathway enzymes;Pseudomonas alcaligenes;Proteome;Stress response proteins
Issue Date: 5-May-2004
Source: ZHAO BING (2004-05-05). Proteome analysis of gentisate-induced responses in Pseudomonas alcaligenes NCIB 9867. ScholarBank@NUS Repository.
Abstract: Pseudomonas alcaligenes NCIB 9867 (designated as P25X) is capable of degradingaromatic hydrocarbons via gentisate pathway. Biochemical characterization of P25Xmutant indicated that it has isofunctional enzymes for the mono- anddioxygenase-catalyzed reactions. One set of the enzymes is constitutively expressedwhereas the other strictly inducible. To date, only the gene encoding theconstitutively-expressed gentisate dioxygenase had been cloned and characterized. Amutant strain of P25X, designated G56, which had the constitutive copy of thegentisate 1,2-dioxygenase gene interrupted by a streptomycin/spectinomycin resistancegene cassette, was found to express gentisate dioxygenase, but only when the cellswere induced by gentisate.The proteome profiles of P25X and mutant G56 grown in the presence and absence ofgentisate were compared after 2D-PAGE. Fifteen distinctive protein spots which wereobserved only in induced cells of P25X and mutant G56 but absent in non-inducedcells of both strains were further analyzed by MALDI-TOF, Q-TOF and N-terminalsequencing. Of the 15 proteins being examined, 13 showed significant sequencesimilarity to the proteins with assigned functions in other microorganisms. Theidentification of protein M5 which showed high homology to a gentisate dioxygenasefrom Ralstonia sp. U2 indicated the putative function of this protein being consistentwith the inducible gentisate 1,2-dioxygenase in P. alcaligenes. In addition, theinduction of stress proteins and other adaptation phenomena were also observed. Theparallel protein analysis of P25X and mutant strain G56 had enabled changes in P.alcaligenes protein expression in response to induction by aromatic hydrocarbons to beunderstood.
URI: http://scholarbank.nus.edu.sg/handle/10635/13768
Appears in Collections:Master's Theses (Open)

Show full item record
Files in This Item:
File Description SizeFormatAccess SettingsVersion 
zhao_bing_Msc_thesis.pdf982.07 kBAdobe PDF

OPEN

NoneView/Download

Page view(s)

194
checked on Dec 11, 2017

Download(s)

267
checked on Dec 11, 2017

Google ScholarTM

Check


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.