Isolation, identification and characterization of 2,2-dichloropropionic acid utilizing bacteria

2,2-dichloropropionic acid (2,2-DCP) is an artificial halogenated compound used as herbicide. A bacterium able to utilize 2,2-DCP as sole carbon source was isolated from soil in Melaka rubber estate. The bacterium was identified as Labrys sp. strain Wy1 using bacterium’s 16S rRNA partial sequence. T...

Full description

Saved in:

Bibliographic Details
Main Author:	Wong, Wen Yong
Format:	Thesis
Language:	English
Published:	2013
Subjects:	QH301 Biology
Online Access:	http://eprints.utm.my/id/eprint/36832/5/WongWenYongMFBSK2013.pdf
Tags:	Add Tag No Tags, Be the first to tag this record!

id	my-utm-ep.36832
record_format	uketd_dc
spelling	my-utm-ep.368322017-07-23T06:24:49Z Isolation, identification and characterization of 2,2-dichloropropionic acid utilizing bacteria 2013-03 Wong, Wen Yong QH301 Biology 2,2-dichloropropionic acid (2,2-DCP) is an artificial halogenated compound used as herbicide. A bacterium able to utilize 2,2-DCP as sole carbon source was isolated from soil in Melaka rubber estate. The bacterium was identified as Labrys sp. strain Wy1 using bacterium’s 16S rRNA partial sequence. The cells doubling time was 34.6 hours in liquid minimal media supplied with 20 mM 2,2-DCP as sole carbon source. Utilization of 2,2-DCP was confirmed by detection of chloride ion released at 0.27 mM. An endophytic bacterium isolated from Axonopus compressus which was identified as Burkholderia cepacia strain Wy5 was also able to utilize 2,2-DCP as sole carbon source. The bacterium has cells doubling time 2.7 hours and chloride ion released was also detected at 47.28 ± 0.25 mM in minimal media contained 20 mM 2,2-DCP. Cell free extract (CFE) of Burkholderia cepacia Wy5 was further characterized due to its higher activity towards 2,2-DCP compared to Labrys sp. Wy1. Dehalogenase found in CFE of Burkholderia cepacia Wy5 has optimal enzyme specific activity at pH8 (0.83 µmol [Cl-] min-1 mg-1) and 40oC (0.78 µmol [Cl-] min-1 mg-1). The dehalogenase was also able to react with other a-haloalkanoic acid including monochloroacetic acid, DL-2-chloropropionic acid and DL-2-bromopropionic acid, but not 3-chloropropionic acid. “Group I” and “Group II” dehalogenase primers were used to amplify dehalogenase gene from both strains Wy1 and Wy5 but only Burkholderia cepacia Wy5 showed positive result. The dehalogenase gene fragment amplified was designated “deh-wy5” and subsequent analysis showed it belongs to Group I dehalogenase. Customized primers based on D,L-dex gene were designed to amplify complete sequence of deh-wy5 due to high similarity between partial sequence of deh-wy5 and D,L-dex. Complete sequence of deh-wy5 was eventually amplified and found to be identical (100%) to D,L-dex. 2013-03 Thesis http://eprints.utm.my/id/eprint/36832/ http://eprints.utm.my/id/eprint/36832/5/WongWenYongMFBSK2013.pdf application/pdf en public masters Universiti Teknologi Malaysia, Faculty of Biosciences and Medical Engineering Faculty of Biosciences and Medical Engineering
institution	Universiti Teknologi Malaysia
collection	UTM Institutional Repository
language	English
topic	QH301 Biology
spellingShingle	QH301 Biology Wong, Wen Yong Isolation, identification and characterization of 2,2-dichloropropionic acid utilizing bacteria
description	2,2-dichloropropionic acid (2,2-DCP) is an artificial halogenated compound used as herbicide. A bacterium able to utilize 2,2-DCP as sole carbon source was isolated from soil in Melaka rubber estate. The bacterium was identified as Labrys sp. strain Wy1 using bacterium’s 16S rRNA partial sequence. The cells doubling time was 34.6 hours in liquid minimal media supplied with 20 mM 2,2-DCP as sole carbon source. Utilization of 2,2-DCP was confirmed by detection of chloride ion released at 0.27 mM. An endophytic bacterium isolated from Axonopus compressus which was identified as Burkholderia cepacia strain Wy5 was also able to utilize 2,2-DCP as sole carbon source. The bacterium has cells doubling time 2.7 hours and chloride ion released was also detected at 47.28 ± 0.25 mM in minimal media contained 20 mM 2,2-DCP. Cell free extract (CFE) of Burkholderia cepacia Wy5 was further characterized due to its higher activity towards 2,2-DCP compared to Labrys sp. Wy1. Dehalogenase found in CFE of Burkholderia cepacia Wy5 has optimal enzyme specific activity at pH8 (0.83 µmol [Cl-] min-1 mg-1) and 40oC (0.78 µmol [Cl-] min-1 mg-1). The dehalogenase was also able to react with other a-haloalkanoic acid including monochloroacetic acid, DL-2-chloropropionic acid and DL-2-bromopropionic acid, but not 3-chloropropionic acid. “Group I” and “Group II” dehalogenase primers were used to amplify dehalogenase gene from both strains Wy1 and Wy5 but only Burkholderia cepacia Wy5 showed positive result. The dehalogenase gene fragment amplified was designated “deh-wy5” and subsequent analysis showed it belongs to Group I dehalogenase. Customized primers based on D,L-dex gene were designed to amplify complete sequence of deh-wy5 due to high similarity between partial sequence of deh-wy5 and D,L-dex. Complete sequence of deh-wy5 was eventually amplified and found to be identical (100%) to D,L-dex.
format	Thesis
qualification_level	Master's degree
author	Wong, Wen Yong
author_facet	Wong, Wen Yong
author_sort	Wong, Wen Yong
title	Isolation, identification and characterization of 2,2-dichloropropionic acid utilizing bacteria
title_short	Isolation, identification and characterization of 2,2-dichloropropionic acid utilizing bacteria
title_full	Isolation, identification and characterization of 2,2-dichloropropionic acid utilizing bacteria
title_fullStr	Isolation, identification and characterization of 2,2-dichloropropionic acid utilizing bacteria
title_full_unstemmed	Isolation, identification and characterization of 2,2-dichloropropionic acid utilizing bacteria
title_sort	isolation, identification and characterization of 2,2-dichloropropionic acid utilizing bacteria
granting_institution	Universiti Teknologi Malaysia, Faculty of Biosciences and Medical Engineering
granting_department	Faculty of Biosciences and Medical Engineering
publishDate	2013
url	http://eprints.utm.my/id/eprint/36832/5/WongWenYongMFBSK2013.pdf
_version_	1747816465354981376

Isolation, identification and characterization of 2,2-dichloropropionic acid utilizing bacteria

Similar Items