Vol. 7(36), pp. 4483-4490, 6 September, 2013
DOI: 10.5897/AJMR12.1690
ISSN 1996-0808 ©2013 Academic Journals
http://www.academicjournals.org/AJMR
African Journal of Microbiology Research
Full Length Research Paper
Isolation, characterization and identification of amylase
producing halothermophilic isolates from
Howz Soltan Lake, Iran
Mahmoudnia Fahimeh
1*
, Bahador Nima
1
and Baserisalehi Majid
2
1
Department of Microbiology, Science and Research Branch, Islamic Azad University, Fars, Iran.
2
Department of Microbiology, Kazeroun Branch, Islamic Azad University, Kazeroun, Iran.
Accepted 2 January, 2013
Halophilic bacteria are a group of salt tolerant microorganisms; the production of enzymes by
halophilic bacteria, due to their activities in harsh environments, is considered as a major field of
investigation. Hence, the present study was conducted for the isolation of halothermophilic bacteria
from Howz Soltan salt lake. For this purpose 40 samples were collected from five regions of the Howz
Soltan lake. The samples were diluted and cultivated on Molten haloid agar (MH) with different salt
concentrations (5-35%). Then the plates were incubated at 35-70°C in aerobic conditions for 7-10 days.
The strains were screened for amylase production based on differences in enzyme production. In total
205 strains were grown on media including 5-15% salt concentrations. Of all, 18 strains were
hyperhalophilic and could grow on media including 20-35% salt concentrations at 40-70°С. Three (3)
isolates were identified as best amylase producers. In addition, the results indicated that maximum
enzyme activity was observed at 48 h (18.145 U/ml). The optimal pH, temperature and NaCl
concentration for amylase production were 7.0-7.5, 40°C and 15% (2.5 M), respectively. Taxonomic and
phylogenetic analysis using 16SrRNA gene sequencing revealed that they belonged to Bacillus
licheniformis and Gracilibacillus while the third isolate was closer to β proteobacterium.
Key words: Halothermophilic bacteria, amylase, Howz Soltan Lake.
INTRODUCTION
Halophilic bacteria are a group of salt tolerant micro-
organisms. These bacteria are divided into three groups:
Slightly, moderately and extremely halophiles (Larsen,
2001). However, survival of halophilic bacteria in the
environment depends on concentration of Na
+
and K
+
, but
compatible solutes in their cells culminate their survival in
ocean, salt lakes and salt fields (Fukushima et al., 2007).
Although production of energy in some halophilic bacteria
depends on their red pigment (bacteriorhodopsin), non
pigmented halophilic bacteria could obtain energy in the
environment with high salt concentrations (Dave et al.,
2006). Nowadays, production of enzymes such as
amylases, proteases and nucleases by halophilic bacteria
due to their activities in harsh environments is considered
as a major field of investigation (Alqueres et al., 2007).
Several reports confirmed that the activity of halophilic
bacterial enzymes remained stable at high salt
concentrations and temperatures (Parakesh and Jaiswal,
2009). However, α- amylase is produced by different
microorganisms viz., Bacillus subtilis, Bacillus
stearothermophilus, Bacillus licheniformis, and Bacillus
amyloliquefaciens, Pseudomonas (Devi et al., 2012;
Bassabrani et al., 2012), Halobacillus sp. (Amoozegar et
al., 2003), Chromohalobacter sp. (Prakesh et al., 2009),
Haloarcula hispanica (Hutcheon et al., 2005), Halomonas
meridiana (Coronado et al., 2000), and Bacillus
*Corresponding author. Email: f_mn_1981@yahoo.com.