Avicelase Production by a Thermophilic Geobacillus stearothermophilus Isolated from Soil using Sugarcane Bagasse
Studies were carried out on the comparative study of the production of Avicelase enzyme using sugarcane bagasse-SCB in two different statuses (i.e. treated and untreated SCB) by thermophilic Geobacillus stearothermophilus at 50ºC. Only four thermophilic bacterial isolates were isolated and assayed for Avicelase production using UntSCB and TSCB. Only one isolate selected as most potent and identified as G. stearothermophilus used in this study. A specific endo-β-1,4-D-glucanase (Avicelase EC 3.2.1.91) was partially purified from a thermophilic bacterial strain was isolated from different soil samples when grown on cellulose enrichment SCB substrate as the sole carbon source. Results shown that G. stearothermophilus was the better Avicelase producer strain. Avicelase had an optimum pH and temperature 7.0 and 50ºC for both UntSCB and TSCB and exhibited good pH stability between "5-8" and "4-9", however, good temperature stability between (30-80ºC) for UntSCB and TSCB, respectively. Other factors affecting the production of Avicelase were compared (i.e. SCB concentration, inoculum size and different incubation periods), all results observed and obtained were revealed that the TSCB was exhibited maximal enzyme activity in comparison with the results obtained from UntSCB, so, the TSCB was enhancing the Avicelase production.
[1] K. Aa, R. Hlengsrud, V. Lindahl, and A. Tronsmo, "Characterization of
production and enzyme properties of an endo-β-1,4-glucanase from
Bacillus subtilis CK-2 isolated from compost soil". Antonie van
Leeuwekoek 66: 319-326, 1994.
[2] C.L. Aguiar, "Biodegradation of the cellulose from sugarcane bagasse
by fungal cellulase". Cienc. Tecnol. Aliment. 2: 117-121, 2001.
[3] E.A. Bayer, J.P. Belaich, Y. ShoHam, R. Lamed, "The cellulosomes:
multienzyme machines for degradation of plant cell wall
polysaccharides". Annu. Rev. Microbiol., 58, 521-554, 2006.
[4] P. Béguin and J.P. Aubert, "The biological degradation of cellulose".
FEMS Microbiol. Rev., 13(1), 25-58, 1994.
[5] M. Bradford, "A rapid and sensitive method for the quantification of
microgram quantities of protein using the principle of protein-dye
binding". Anal Biochem; 72: 248-54, 1976.
[6] K. Bronnenmeier, and W.L. staudenbauer, "Cellulose hydrolysis by a
highly thermostable endo-1,4-β-glucanase (Avicelase I) from
Clostridium stercorarium". Enzyme Microb. Technol. 12, 431-436,
1990.
[7] K. Cavedon, S.B. Leschine, and E. Canale-Parola, "Characterization of
the extracellular cellulase from a mesophilic Clostridium (strain C7)" J.
Bacteriol., 172: 4222-4230, 1990.
[8] K.Y. Chan, and K.S. Au, "Studies on cellulase production by a Bacillus
subtilis" Antonie Van Leeuwenhoek. 53:125-136, 1987.
[9] R.J. Chr├│st, "Microbial ectoenzymes in aquatic environments" In:
Overbeck, J., Chr├│st, R.J. (eds). Aquatic microbial ecology: biochemical
and molecular approaches. Springer-Verlag, New York, USA, p. 47-78,
1990.
[10] M.B. Cunha-Santino, and Jr.I. Bianchini, "Cellulase activities during
decomposition of a submerged aquatic macrophyte (Utricularia
breviscapa): A microcosm assay" Brazillian Journal of
Microbiology.38:230-236, 2007.
[11] M. Desvaux, "The cellulosome of Clostridium cellulolyticum" Enzyme
Microb. Technol., 37, 373-385, 2005.
[12] M. Dixon, and E.C. Webb, "Enzymes" 2nd Edit. Academic Press Inc.
New York, 1964.
[13] G. Gomori, "Preparation Of Buffers For Use In Enzyme Active Studies"
Method In Enzymol. I, 138-146. Academic Press, London, 1955.
[14] K. Horikoshi, M. Nakao, Y. Kurono, and N. Sashihara, "Cellulases of an
alkalophilic Bacillus strain isolated from soil" Can. J. Microbiol. 30:
774-779, 1984.
[15] H. Jorgensen, T. Eriksson, J. Borjesson, F. Tjerneld, and L. Olsson,
"Purification and characterization of five cellulases and xylanase from
Penicilliul brasilianum IBI 20888" Enzyme and Microbial Technology.
32: 851-861, 2003.
[16] A. Kyriacou, C.R. MacKenzie, and R.J. Neufeld, "Detection and
characterization of the specific and nonspecific endoglucanases of Trichoderma reesei: Evidence demonstrating endoglucanase activity by
cellobiohydrolase II" Enzyme Microb. Technol., 9: 25-32, 1987.
[17] R. Muthuvelayudham, and T. Viruthagiri, "Biodegradation of sugarcane
bagasse using T. reesei cellulase protein" Chemical Engineering
Congress, 2005.
[18] R. Muthuvelayudham, and T. Viruthagiri, "Fermentative production and
kinetics of cellulose protein on Trichoderma reesei using sugarcane
bagasse and rice straw" African Journal of Biotechnology. Vol. 5 (20),
pp. 1873-1881, 2006.
[19] T.K. Ng, and J.G. Zeikus, "Comparison of extracellular cellulase
activities of Clostridium thermocellum LQR1 and Trichoderma reesei
QM9414" Appl. Environ. Microbiol. 42:231-240, 1981.
[20] H. Okoshi, O. Katsukata, S. Shikata, K. Oshino, S. Kawai, and S. Ito,
"Purification and characterization of multiple carboxymethylcellulases
from Bacillus sp. KSM-522" Agrc Biol Chem. 54: 83-89, 1990.
[21] L. Olsson, TMIE Christensen, K.P. Hasen, E.A. Palmqvist, "Influence of
the carbon source on production of cellulases, hemicellulases and
pectinases by Trichoderma reesei Rut C-30" Enzyme and Microbial
Technol. 33: 612-619, 2003.
[22] K. Ozaki, and S. Ito, "Purification and properties of an acid endo-β-1,4-
glucanase from Bacillus sp. KSM-330" J Gen Appl Microbiol. 137: 41-
48, 1991.
[23] P.H. Reynolds, C.H. Sissons, R.M. Daniel, H.W. Morgan, "Comparison
of cellulolytic activities in Clostridium thermocellum and three
thermophilic, cellulolytic anaerobes" Appl. Environ. Microbiol. 51: 12-
17, 1986.
[24] L.M. Robson, and G.H. Chambliss, "Characterization of the cellulolytic
activity of a Bacillus isolate" Appl. Environ. Microbiol. 47: 1039-1046,
1984.
[25] J.N. Saddler, A.W. Khan, S.M. Martin, "Regulation of cellulase
synthesis in Acetivibrio cellulolyticus" Microbios, 28, 97-106, 1980.
[26] M. Schallmey, A. Singh, and O.P. Ward, "Developments in the use of
Bacillus species for industrial production" Can J Microbiol, 50: 1-17,
2004.
[27] G. Schimid, and C. Wandrey, "Evidence for lack of cellobiohydrolase
activity in the cellulase sysem of Trichoderma reesei QM 9414" J.
Biotechnol. 14: 393-410, 1990.
[28] W.H. Schwarz, "The cellulosome and cellulose degradation by anaerobic
bacteria" Appl. Microbiol. Biotechnol., 56, 634-649, 2001.
[29] J. Stahlberg, G. Johansson, and G. Pettersson, "Trichoderma reesei has
no true exo-cellulase; All intact and truncated cellulases produce new
reducing end groups on cellulose" Biochim. Biophys. Acta ., 1157: 107-
113, 1993.
[30] P.E. Stukus, "Investigating Microbiology a laboratory Manual for
general Microbiology. Saunders College Publishing" Harcourt Brace
College Publishers, 1997.
[31] J. Zemek, J. Augustin, R. Borriss, D. Kuniak, M. Svabova, Z. Pacova,
"Polysaccharide-hydrolyzing enzymes in the genus Bacillus" Folia
Microbiol 26: 403-407, 1981.
[1] K. Aa, R. Hlengsrud, V. Lindahl, and A. Tronsmo, "Characterization of
production and enzyme properties of an endo-β-1,4-glucanase from
Bacillus subtilis CK-2 isolated from compost soil". Antonie van
Leeuwekoek 66: 319-326, 1994.
[2] C.L. Aguiar, "Biodegradation of the cellulose from sugarcane bagasse
by fungal cellulase". Cienc. Tecnol. Aliment. 2: 117-121, 2001.
[3] E.A. Bayer, J.P. Belaich, Y. ShoHam, R. Lamed, "The cellulosomes:
multienzyme machines for degradation of plant cell wall
polysaccharides". Annu. Rev. Microbiol., 58, 521-554, 2006.
[4] P. Béguin and J.P. Aubert, "The biological degradation of cellulose".
FEMS Microbiol. Rev., 13(1), 25-58, 1994.
[5] M. Bradford, "A rapid and sensitive method for the quantification of
microgram quantities of protein using the principle of protein-dye
binding". Anal Biochem; 72: 248-54, 1976.
[6] K. Bronnenmeier, and W.L. staudenbauer, "Cellulose hydrolysis by a
highly thermostable endo-1,4-β-glucanase (Avicelase I) from
Clostridium stercorarium". Enzyme Microb. Technol. 12, 431-436,
1990.
[7] K. Cavedon, S.B. Leschine, and E. Canale-Parola, "Characterization of
the extracellular cellulase from a mesophilic Clostridium (strain C7)" J.
Bacteriol., 172: 4222-4230, 1990.
[8] K.Y. Chan, and K.S. Au, "Studies on cellulase production by a Bacillus
subtilis" Antonie Van Leeuwenhoek. 53:125-136, 1987.
[9] R.J. Chr├│st, "Microbial ectoenzymes in aquatic environments" In:
Overbeck, J., Chr├│st, R.J. (eds). Aquatic microbial ecology: biochemical
and molecular approaches. Springer-Verlag, New York, USA, p. 47-78,
1990.
[10] M.B. Cunha-Santino, and Jr.I. Bianchini, "Cellulase activities during
decomposition of a submerged aquatic macrophyte (Utricularia
breviscapa): A microcosm assay" Brazillian Journal of
Microbiology.38:230-236, 2007.
[11] M. Desvaux, "The cellulosome of Clostridium cellulolyticum" Enzyme
Microb. Technol., 37, 373-385, 2005.
[12] M. Dixon, and E.C. Webb, "Enzymes" 2nd Edit. Academic Press Inc.
New York, 1964.
[13] G. Gomori, "Preparation Of Buffers For Use In Enzyme Active Studies"
Method In Enzymol. I, 138-146. Academic Press, London, 1955.
[14] K. Horikoshi, M. Nakao, Y. Kurono, and N. Sashihara, "Cellulases of an
alkalophilic Bacillus strain isolated from soil" Can. J. Microbiol. 30:
774-779, 1984.
[15] H. Jorgensen, T. Eriksson, J. Borjesson, F. Tjerneld, and L. Olsson,
"Purification and characterization of five cellulases and xylanase from
Penicilliul brasilianum IBI 20888" Enzyme and Microbial Technology.
32: 851-861, 2003.
[16] A. Kyriacou, C.R. MacKenzie, and R.J. Neufeld, "Detection and
characterization of the specific and nonspecific endoglucanases of Trichoderma reesei: Evidence demonstrating endoglucanase activity by
cellobiohydrolase II" Enzyme Microb. Technol., 9: 25-32, 1987.
[17] R. Muthuvelayudham, and T. Viruthagiri, "Biodegradation of sugarcane
bagasse using T. reesei cellulase protein" Chemical Engineering
Congress, 2005.
[18] R. Muthuvelayudham, and T. Viruthagiri, "Fermentative production and
kinetics of cellulose protein on Trichoderma reesei using sugarcane
bagasse and rice straw" African Journal of Biotechnology. Vol. 5 (20),
pp. 1873-1881, 2006.
[19] T.K. Ng, and J.G. Zeikus, "Comparison of extracellular cellulase
activities of Clostridium thermocellum LQR1 and Trichoderma reesei
QM9414" Appl. Environ. Microbiol. 42:231-240, 1981.
[20] H. Okoshi, O. Katsukata, S. Shikata, K. Oshino, S. Kawai, and S. Ito,
"Purification and characterization of multiple carboxymethylcellulases
from Bacillus sp. KSM-522" Agrc Biol Chem. 54: 83-89, 1990.
[21] L. Olsson, TMIE Christensen, K.P. Hasen, E.A. Palmqvist, "Influence of
the carbon source on production of cellulases, hemicellulases and
pectinases by Trichoderma reesei Rut C-30" Enzyme and Microbial
Technol. 33: 612-619, 2003.
[22] K. Ozaki, and S. Ito, "Purification and properties of an acid endo-β-1,4-
glucanase from Bacillus sp. KSM-330" J Gen Appl Microbiol. 137: 41-
48, 1991.
[23] P.H. Reynolds, C.H. Sissons, R.M. Daniel, H.W. Morgan, "Comparison
of cellulolytic activities in Clostridium thermocellum and three
thermophilic, cellulolytic anaerobes" Appl. Environ. Microbiol. 51: 12-
17, 1986.
[24] L.M. Robson, and G.H. Chambliss, "Characterization of the cellulolytic
activity of a Bacillus isolate" Appl. Environ. Microbiol. 47: 1039-1046,
1984.
[25] J.N. Saddler, A.W. Khan, S.M. Martin, "Regulation of cellulase
synthesis in Acetivibrio cellulolyticus" Microbios, 28, 97-106, 1980.
[26] M. Schallmey, A. Singh, and O.P. Ward, "Developments in the use of
Bacillus species for industrial production" Can J Microbiol, 50: 1-17,
2004.
[27] G. Schimid, and C. Wandrey, "Evidence for lack of cellobiohydrolase
activity in the cellulase sysem of Trichoderma reesei QM 9414" J.
Biotechnol. 14: 393-410, 1990.
[28] W.H. Schwarz, "The cellulosome and cellulose degradation by anaerobic
bacteria" Appl. Microbiol. Biotechnol., 56, 634-649, 2001.
[29] J. Stahlberg, G. Johansson, and G. Pettersson, "Trichoderma reesei has
no true exo-cellulase; All intact and truncated cellulases produce new
reducing end groups on cellulose" Biochim. Biophys. Acta ., 1157: 107-
113, 1993.
[30] P.E. Stukus, "Investigating Microbiology a laboratory Manual for
general Microbiology. Saunders College Publishing" Harcourt Brace
College Publishers, 1997.
[31] J. Zemek, J. Augustin, R. Borriss, D. Kuniak, M. Svabova, Z. Pacova,
"Polysaccharide-hydrolyzing enzymes in the genus Bacillus" Folia
Microbiol 26: 403-407, 1981.
@article{"International Journal of Biological, Life and Agricultural Sciences:64619", author = "E. A. Makky", title = "Avicelase Production by a Thermophilic Geobacillus stearothermophilus Isolated from Soil using Sugarcane Bagasse", abstract = "Studies were carried out on the comparative study of the production of Avicelase enzyme using sugarcane bagasse-SCB in two different statuses (i.e. treated and untreated SCB) by thermophilic Geobacillus stearothermophilus at 50ºC. Only four thermophilic bacterial isolates were isolated and assayed for Avicelase production using UntSCB and TSCB. Only one isolate selected as most potent and identified as G. stearothermophilus used in this study. A specific endo-β-1,4-D-glucanase (Avicelase EC 3.2.1.91) was partially purified from a thermophilic bacterial strain was isolated from different soil samples when grown on cellulose enrichment SCB substrate as the sole carbon source. Results shown that G. stearothermophilus was the better Avicelase producer strain. Avicelase had an optimum pH and temperature 7.0 and 50ºC for both UntSCB and TSCB and exhibited good pH stability between "5-8" and "4-9", however, good temperature stability between (30-80ºC) for UntSCB and TSCB, respectively. Other factors affecting the production of Avicelase were compared (i.e. SCB concentration, inoculum size and different incubation periods), all results observed and obtained were revealed that the TSCB was exhibited maximal enzyme activity in comparison with the results obtained from UntSCB, so, the TSCB was enhancing the Avicelase production.
", keywords = "Geobacillus stearothermophilus, Avicelase, Sugarcane bagasse", volume = "3", number = "9", pages = "492-5", }
