African Journal of Microbiology Research Vol. 5(28), pp. 4987-4992, 30 November, 2011
Available online at http://www.academicjournals.org/AJMR
ISSN 1996-0808 ©2011 Academic Journals
DOI: 10.5897/AJMR11.505
Full Length Research Paper
Characterisation of Bacillus strains from volcanic area
Gunung Sibayak able to degrade 2,2-dichloropropionic
acid
Damia Diyana Roslan
1
, Ronnie G. Gicana
2
, Robert J. Lamis
2
and Fahrul Huyop
1
*
1
Department of Industrial Biotechnology, Faculty of Biosciences and Bioengineering, University Teknologi Malaysia,
81310 Skudai, Johor, Malaysia.
2
College of Arts and Sciences, University of Negros Occidental-Recoletos, 6100 Bacolod City, Philippines.
Accepted 8 November, 2011
Halogenated compounds are mainly found in agricultural area. The problem arose because these
compounds are toxic to human and are persistent in environment. The aim of the present study was to
isolate 2,2-dichloropropionic acid (2,2-DCP) degrading bacteria that can grow at higher temperature
rather than at normal 25 to 30°C temperature. A soil bacterium was isolated from the surrounding
volcanic area at the feet of Gunung Sibayak, Indonesia using streak plate method. The sample was
repeatedly streaked onto 20 mM of 2,2-DCP to get a pure culture. The culture was incubated for 2 days at
30, 40 and 60°C incubator shaker aerobically. The isolated bacterium grew best at 40°C but failed to grow
at 60°C. Based on partial 16S rDNA gene sequence, the isolated strain GS1 showed high sequence
identity to Bacillus megaterium strain 0 to 2, whereas, using biochemical tests suggested that the genus
belongs to Bacillus sp. as expected. Strain GS1 was a Gram positive, rod in shape and produced spore.
The results also suggested that, strain GS1 could degrade 20 mM (2,2-DCP) based on growth
experiment. In conclusion, the molecular method identified Bacillus genus showing no discrepencies
from the biochemical test results. This is the first report of Bacillus strains associated with 2,2-DCP
degradation using above normal temperature for growth.
Key words: 2,2-Dichloropropionic, Bacillus strain GS1, degradation, 16S rDNA and pollutant.
INTRODUCTION
Halogenated compounds constitute the most important
class of xenobiotic which mostly make up pollution.
Some of these compounds are very toxic and cause
enormous problems to human health and to the
environment (van Pee and Unversucht, 2003). Several
microorganisms reported to utilize 2-halogenated
alkanoic acids as a sole carbon source have been
isolated (Jing and Huyop, 2008; Ismail et al., 2008; Jing
et al., 2008; Thasif et al., 2009; Mesri et al., 2009). The
bacterial degradation of 2-haloalkanoic acids is attributed
to an enzyme, dehalogenase, which catalyzes the
hydrolytic removal of halogens from 2-haloalkanoic acids
*Corresponding author. E-mail: fzhutm@gmail.com. Tel: +607
5534556. Fax: +607 5531112.
(Kurihara, 2011). Previous studies also described cloning
of dehalogenase in E. coli system for dehalogenase
enzyme characterization (Cairns et al., 1996; Stringfellow
et al., 1997; Yusn and Huyop, 2009; Huyop et al., 2008).
Some halogenated chemicals are easily biodegradable
for example 2-chloropropionate that can be enriched from
almost any oil sample (Leisinger, 1996). This study
initially screened the ability of bacteria in soil sample from
volcanic area of Gunung Sibayak to grow at high
temperature in the presence of α-haloalkanoate.
Research interests are geared towards understanding
microbial growth at higher temperature as enzymes from
thermophillic organisms often show stability to a number
of extrinsic factors such as temperatures and organic
solvents. These properties make them useful for
industrial biocatalyse (Rye et al., 2009).
In this paper we report the isolation and identification of