Identification of Cellulose-Hydrolytic Thermophiles Isolated from Sg. Klah Hot Spring Based On 16S rDNA Gene Sequence
In this study, six bacterial isolates of a slightly
thermophilic organism from the Sg. Klah hot spring, Malaysia were
successfully isolated and designated as M7T55D1, M7T55D2,
M7T55D3, M7T53D1, M7T53D2 and M7T53D3 respectively. The
bacterial isolates were screened for their cellulose hydrolytic ability
on Carboxymethlycellulose agar medium. The isolated bacterial
strains were identified morphologically, biochemically and
molecularly with the aid of 16S rDNA sequencing. All of the bacteria
showed their optimum growth at a slightly alkaline pH of 7.5 with a
temperature of 55°C. All strains were Gram-negative, non-spore
forming type, strictly aerobic, catalase-positive and oxidase-positive
with the ability to produce thermostable cellulase. Based on BLASTn
results, bacterial isolates of M7T55D2 and M7T53D1 gave the
highest homology (97%) with similarity to Tepidimonas ignava while
isolates M7T55D1, M7T55D3, M7T53D2 and M7T53D3 showed
their closest homology (97%-98%) with Tepidimonas thermarum.
These cellulolytic thermophiles might have a commercial potential to
produce valuable thermostable cellulase.
[1] H. Zuridah, N. Norazwin, M. Siti Aisyah, M. N. A. Fakhruzzaman and
N. A. Zeenathul, "Identification of lipase producing thermophilic
bacteria from Malaysian hot springs,” African Journal of Microbiology
Research, vol. 5, pp. 3569-3573, 2011.
[2] Haki, G.D. and Rakshit, S.K. (2003). Development in industrially
important thermostable enzyme: A review. Biosource Technology, 89:
17-34.
[3] D. Gaur, K. J. Pankaj, S. S. Yamini and B. Vivek, "Estimation of
extracellular lipolytic enzyme activity by thermophillic Bacillus sp.
isolated from Arid and semi-Arid region of Rajasthan, India,” Journal of
Microbiology Biotechnology and Food Sciences, vol. 2 (2), pp. 619-633,
2012.
[4] A. Somen and C. Anita (2012). "Bioprospecting thermophiles for
cellulase production,” Brazilian Journal of Microbiology, vol. 43, pp.
164 – 187, 2002.
[5] V. Liisa, A. Marika, P. Terhi, V. Jari and S. Matti, "Thermostable
enzymes in lignocellulose hydrolysis,” Journal of Advance Biochem
Engineering/Biotechnology, vol 108, pp. 121–145, 2007.
[6] C. Vielle and G. J. Zeikus, "Hyperthermophilic enzymes: sources, uses
and molecular mechanisms for thermostability,” Microbiology and
Molecular Biology Reviews, vol. 65, pp. 1-43, 2001.
[7] M. Bhat, "Cellulases and related enzymes in biotechnology,”
Biotechnol. Adv., vol 18, pp. 355–383, 2000.
[8] R. D., Firn, "Bioprospecting-Why is it so unrewarding?,” Biodiversity
and Conservation, vol. 12, pp. 207-216, 2003.
[9] X. Li, H. Yang, B. Roy, D. Wang, W. Yue, L. Jiang, E. Park and Y.
Miao, "The most stirring technology in future: Cellulase enzyme and
biomass utilization,” African Journal of Technology, vol. 8 (11), pp.
2418-2422, 2009.
[10] S. S. I., Abdelnasser and I. E., Ahmed, "Isolation and identification of
new cellulase producing thermophilic bacteria from an Egyptian hot
springs and some properties of the crude enzyme,” Australian Journal of
Basic and Applied Sciences, vol. 1(4), pp. 473-478, 2007.
[11] H. Yuichi, Y. Hiroe, O. Moriya and K. Toshiaki, ”Evaluation of primers
and PCR condition for the analysis of 16S rDNA genes from a natural
environment,” FEMS Microbiology Letters, vol. 221, pp. 299-304, 2003.
[12] L. Albuquerque, I. Tiago, A. Verissimo and M. S. Costa, "Tepidimonas
thermarum sp. nov., a new slightly thermophilic betaproteobacterium
isolated from the Elisenquelle in Aachen and emended description of the
genus Tepidimonas,” Systematic and Applied Microbiology, vol. 29, pp.
450 - 456, 2005.
[13] M. Freitas, F. R. Rainey, M. F. Nobre, A. J. D. Silvestre and M. S. da
Costa, "Tepidimonas aquatica sp. nov., a new slightly thermophilic b-
Proteobacterium isolated from a hot water tank,” System, Issue Series
Title: Appl. Microbiol., vol. 26, pp. 376–381, 2003.
[14] T. L. Chen, Y. J. Chou, W. M. Chen, B. Arun, C. C. Young,
"Tepidimonas taiwanensis sp. nov., a novel alkalineprotease- producing
bacterium isolated from a hot spring,” Application Environment
Microbiology, vol. 10, pp. 35 – 40, 2005.
[15] C. Moreira, F. A. Rainey, M. F. Nobre, M. T. Silva, M. S. da Costa,
"Tepidimonas ignava gen. nov., sp. nov., a new chemolithoheterotrophic
and slightly thermophilic member of the b-Proteobacteria,” Int. J. Syst.
Bacteriol. vol. 50, pp. 735–742, 2000.
[1] H. Zuridah, N. Norazwin, M. Siti Aisyah, M. N. A. Fakhruzzaman and
N. A. Zeenathul, "Identification of lipase producing thermophilic
bacteria from Malaysian hot springs,” African Journal of Microbiology
Research, vol. 5, pp. 3569-3573, 2011.
[2] Haki, G.D. and Rakshit, S.K. (2003). Development in industrially
important thermostable enzyme: A review. Biosource Technology, 89:
17-34.
[3] D. Gaur, K. J. Pankaj, S. S. Yamini and B. Vivek, "Estimation of
extracellular lipolytic enzyme activity by thermophillic Bacillus sp.
isolated from Arid and semi-Arid region of Rajasthan, India,” Journal of
Microbiology Biotechnology and Food Sciences, vol. 2 (2), pp. 619-633,
2012.
[4] A. Somen and C. Anita (2012). "Bioprospecting thermophiles for
cellulase production,” Brazilian Journal of Microbiology, vol. 43, pp.
164 – 187, 2002.
[5] V. Liisa, A. Marika, P. Terhi, V. Jari and S. Matti, "Thermostable
enzymes in lignocellulose hydrolysis,” Journal of Advance Biochem
Engineering/Biotechnology, vol 108, pp. 121–145, 2007.
[6] C. Vielle and G. J. Zeikus, "Hyperthermophilic enzymes: sources, uses
and molecular mechanisms for thermostability,” Microbiology and
Molecular Biology Reviews, vol. 65, pp. 1-43, 2001.
[7] M. Bhat, "Cellulases and related enzymes in biotechnology,”
Biotechnol. Adv., vol 18, pp. 355–383, 2000.
[8] R. D., Firn, "Bioprospecting-Why is it so unrewarding?,” Biodiversity
and Conservation, vol. 12, pp. 207-216, 2003.
[9] X. Li, H. Yang, B. Roy, D. Wang, W. Yue, L. Jiang, E. Park and Y.
Miao, "The most stirring technology in future: Cellulase enzyme and
biomass utilization,” African Journal of Technology, vol. 8 (11), pp.
2418-2422, 2009.
[10] S. S. I., Abdelnasser and I. E., Ahmed, "Isolation and identification of
new cellulase producing thermophilic bacteria from an Egyptian hot
springs and some properties of the crude enzyme,” Australian Journal of
Basic and Applied Sciences, vol. 1(4), pp. 473-478, 2007.
[11] H. Yuichi, Y. Hiroe, O. Moriya and K. Toshiaki, ”Evaluation of primers
and PCR condition for the analysis of 16S rDNA genes from a natural
environment,” FEMS Microbiology Letters, vol. 221, pp. 299-304, 2003.
[12] L. Albuquerque, I. Tiago, A. Verissimo and M. S. Costa, "Tepidimonas
thermarum sp. nov., a new slightly thermophilic betaproteobacterium
isolated from the Elisenquelle in Aachen and emended description of the
genus Tepidimonas,” Systematic and Applied Microbiology, vol. 29, pp.
450 - 456, 2005.
[13] M. Freitas, F. R. Rainey, M. F. Nobre, A. J. D. Silvestre and M. S. da
Costa, "Tepidimonas aquatica sp. nov., a new slightly thermophilic b-
Proteobacterium isolated from a hot water tank,” System, Issue Series
Title: Appl. Microbiol., vol. 26, pp. 376–381, 2003.
[14] T. L. Chen, Y. J. Chou, W. M. Chen, B. Arun, C. C. Young,
"Tepidimonas taiwanensis sp. nov., a novel alkalineprotease- producing
bacterium isolated from a hot spring,” Application Environment
Microbiology, vol. 10, pp. 35 – 40, 2005.
[15] C. Moreira, F. A. Rainey, M. F. Nobre, M. T. Silva, M. S. da Costa,
"Tepidimonas ignava gen. nov., sp. nov., a new chemolithoheterotrophic
and slightly thermophilic member of the b-Proteobacteria,” Int. J. Syst.
Bacteriol. vol. 50, pp. 735–742, 2000.
@article{"International Journal of Biological, Life and Agricultural Sciences:68072", author = "M. J. Norashirene and Y. Zakiah and S. Nurdiana and I. Nur Hilwani and M. H. Siti Khairiyah and M. J. Muhamad Arif", title = "Identification of Cellulose-Hydrolytic Thermophiles Isolated from Sg. Klah Hot Spring Based On 16S rDNA Gene Sequence", abstract = "In this study, six bacterial isolates of a slightly
thermophilic organism from the Sg. Klah hot spring, Malaysia were
successfully isolated and designated as M7T55D1, M7T55D2,
M7T55D3, M7T53D1, M7T53D2 and M7T53D3 respectively. The
bacterial isolates were screened for their cellulose hydrolytic ability
on Carboxymethlycellulose agar medium. The isolated bacterial
strains were identified morphologically, biochemically and
molecularly with the aid of 16S rDNA sequencing. All of the bacteria
showed their optimum growth at a slightly alkaline pH of 7.5 with a
temperature of 55°C. All strains were Gram-negative, non-spore
forming type, strictly aerobic, catalase-positive and oxidase-positive
with the ability to produce thermostable cellulase. Based on BLASTn
results, bacterial isolates of M7T55D2 and M7T53D1 gave the
highest homology (97%) with similarity to Tepidimonas ignava while
isolates M7T55D1, M7T55D3, M7T53D2 and M7T53D3 showed
their closest homology (97%-98%) with Tepidimonas thermarum.
These cellulolytic thermophiles might have a commercial potential to
produce valuable thermostable cellulase.
", keywords = "Cellulase, Cellulolytic, Thermophiles, 16S rDNA
Gene.", volume = "8", number = "9", pages = "1041-4", }
