Isolation of local bacterial capable of degrading halogenated compounds and analysis of putative haloacid permease gene

3-chloropropionic acid and 2,2-dichloropropionic acid are synthetic halogenated compounds used in herbicide. A bacterium isolated from a soil sample and characterised as Rhodococcus sp. by 16S rRNA analysis, was able to degrade and utilised 3-chloropropionate as the sole source of carbon and energy....

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Main Author:	Ng, Hong Jing
Format:	Thesis
Language:	English
Published:	2007
Subjects:	QH301 Biology
Online Access:	http://eprints.utm.my/id/eprint/6798/1/NgHongJingMFS2007.pdf
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spelling	my-utm-ep.67982018-08-30T08:03:52Z Isolation of local bacterial capable of degrading halogenated compounds and analysis of putative haloacid permease gene 2007-02 Ng, Hong Jing QH301 Biology 3-chloropropionic acid and 2,2-dichloropropionic acid are synthetic halogenated compounds used in herbicide. A bacterium isolated from a soil sample and characterised as Rhodococcus sp. by 16S rRNA analysis, was able to degrade and utilised 3-chloropropionate as the sole source of carbon and energy. This was supported by the ability of the bacterium to grow on 20 mM 3-chloropropionate with a doubling time of 11.72 hours. The utilisation of 3-chloropropionate was also confirmed by detection of 3-chloropropionate depletion in the medium using HPLC. Cell free extract of Rhodococcus sp. had an enzyme specific activity of 0.013 Âµmol Cl-/min/mg protein towards 3-chloropropionate. Another bacterium isolated from the same soil sample and identified as Methylobacterium sp. by 16S rRNA analysis was found to be able to degrade 2,2-dichloropropionate. The bacterium grew in 20mM 2,2-dichloropropionate minimal medium with a doubling time of 20.32 hours. Degradation of 2,2-dichloropropionate was further confirmed by detection of 2,2- dichloropropionate depletion in growth medium by HPLC. Cell free extract prepared from the cell showed 0.039 Âµmol Cl-/min/mg protein specific activity towards 2,2-dichloropropionate. A putative haloacid permease gene (dehrP) from Rhizobium sp. was subcloned into Novagen pET 43.1a plasmid. The newly constructed plasmid was designated as pHJ. The cloned gene was sequenced and analysed using various online analysis tools. DehrP has a calculated molecular weight of 45 kDa and an isoelectric point of 9.78. The nucleotide sequence of dehrP showed significant homology (86%) with the putative mono-chloropropionic acid permease from Agrobacterium sp. NHG3 and 62 % homology with the haloacid specific transferase from Burkholderia sp. 2007-02 Thesis http://eprints.utm.my/id/eprint/6798/ http://eprints.utm.my/id/eprint/6798/1/NgHongJingMFS2007.pdf application/pdf en public http://dms.library.utm.my:8080/vital/access/manager/Repository/vital:62525 masters Universiti Teknologi Malaysia, Faculty of Science Faculty of Science
institution	Universiti Teknologi Malaysia
collection	UTM Institutional Repository
language	English
topic	QH301 Biology
spellingShingle	QH301 Biology Ng, Hong Jing Isolation of local bacterial capable of degrading halogenated compounds and analysis of putative haloacid permease gene
description	3-chloropropionic acid and 2,2-dichloropropionic acid are synthetic halogenated compounds used in herbicide. A bacterium isolated from a soil sample and characterised as Rhodococcus sp. by 16S rRNA analysis, was able to degrade and utilised 3-chloropropionate as the sole source of carbon and energy. This was supported by the ability of the bacterium to grow on 20 mM 3-chloropropionate with a doubling time of 11.72 hours. The utilisation of 3-chloropropionate was also confirmed by detection of 3-chloropropionate depletion in the medium using HPLC. Cell free extract of Rhodococcus sp. had an enzyme specific activity of 0.013 Âµmol Cl-/min/mg protein towards 3-chloropropionate. Another bacterium isolated from the same soil sample and identified as Methylobacterium sp. by 16S rRNA analysis was found to be able to degrade 2,2-dichloropropionate. The bacterium grew in 20mM 2,2-dichloropropionate minimal medium with a doubling time of 20.32 hours. Degradation of 2,2-dichloropropionate was further confirmed by detection of 2,2- dichloropropionate depletion in growth medium by HPLC. Cell free extract prepared from the cell showed 0.039 Âµmol Cl-/min/mg protein specific activity towards 2,2-dichloropropionate. A putative haloacid permease gene (dehrP) from Rhizobium sp. was subcloned into Novagen pET 43.1a plasmid. The newly constructed plasmid was designated as pHJ. The cloned gene was sequenced and analysed using various online analysis tools. DehrP has a calculated molecular weight of 45 kDa and an isoelectric point of 9.78. The nucleotide sequence of dehrP showed significant homology (86%) with the putative mono-chloropropionic acid permease from Agrobacterium sp. NHG3 and 62 % homology with the haloacid specific transferase from Burkholderia sp.
format	Thesis
qualification_level	Master's degree
author	Ng, Hong Jing
author_facet	Ng, Hong Jing
author_sort	Ng, Hong Jing
title	Isolation of local bacterial capable of degrading halogenated compounds and analysis of putative haloacid permease gene
title_short	Isolation of local bacterial capable of degrading halogenated compounds and analysis of putative haloacid permease gene
title_full	Isolation of local bacterial capable of degrading halogenated compounds and analysis of putative haloacid permease gene
title_fullStr	Isolation of local bacterial capable of degrading halogenated compounds and analysis of putative haloacid permease gene
title_full_unstemmed	Isolation of local bacterial capable of degrading halogenated compounds and analysis of putative haloacid permease gene
title_sort	isolation of local bacterial capable of degrading halogenated compounds and analysis of putative haloacid permease gene
granting_institution	Universiti Teknologi Malaysia, Faculty of Science
granting_department	Faculty of Science
publishDate	2007
url	http://eprints.utm.my/id/eprint/6798/1/NgHongJingMFS2007.pdf
_version_	1747814688591183872

Isolation of local bacterial capable of degrading halogenated compounds and analysis of putative haloacid permease gene

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