Please use this identifier to cite or link to this item: http://scholarbank.nus.edu.sg/handle/10635/16964
Title: Genetic and biochemical characterization of unknown XlnD and HbzB from Pseudomonas Alcaligenes NCIMB 9867
Authors: GAO XIAOLI
Keywords: gentisate pathway, Pseudomonas alcaligenes NCIMB 9867, 3-hydroxybenzoate 6-hydroxylase, gentisate 1,2-dioxygenase
Issue Date: 20-May-2005
Source: GAO XIAOLI (2005-05-20). Genetic and biochemical characterization of unknown XlnD and HbzB from Pseudomonas Alcaligenes NCIMB 9867. ScholarBank@NUS Repository.
Abstract: Pseudomonas alcaligenes NCIMB 9867 (strain P25X) produces isofunctional enzymes of the gentisate pathway that enable the degradation of xylenols and cresols. In this study, we investigated the function of xlnD, overexpressed it, purified and characterized the protein XlnD from E. coli BL21. Enzyme assays showed that XlnD was functioning as a 3-hydroxybenzoate 6-hydroxylase. A P25X xlnD knockout mutant, strain G50, was constructed and in this mutant, 6-hydroxylase activity could only be detected when cells were grown in either 3-hydroxybenzoate or gentisate, thus indicating the presence of another 6-hydroxylase (6-hydroxylase II) in strain P25X which was strictly inducible by 3-hydroxybenzoate. Strain G50 lost the ability to grow on 2,5-xylenol, 3,5-xylenol and 3-hydroxy-4-methylbenzoate and this ability was restored when xlnD was provided on a separate pRK415-derived plasmid. Sequence analysis of the gene hbzB indicated that it encodes gentisate 1,2-dioxygenase II. Expression, purification and characterization of gentisate 1,2-dioxygenase II was also reported in this work.
URI: http://scholarbank.nus.edu.sg/handle/10635/16964
Appears in Collections:Master's Theses (Open)

Show full item record
Files in This Item:
File Description SizeFormatAccess SettingsVersion 
GaoXL thesis part 1.pdf6.81 kBAdobe PDF

OPEN

NoneView/Download
GaoXL thesis part 2.pdf29.28 kBAdobe PDF

OPEN

NoneView/Download
GaoXL thesis part 3.pdf915.95 kBAdobe PDF

OPEN

NoneView/Download
GaoXL thesis spine.pdf9.94 kBAdobe PDF

OPEN

NoneView/Download

Page view(s)

298
checked on Dec 11, 2017

Download(s)

1,173
checked on Dec 11, 2017

Google ScholarTM

Check


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