Proteolytic Dedradation of Anchovy (Spolephorus spp.) Proteins by Halophilic Proteinase from Halobacillus sp. SR5-3
The halophilic proteinase showed a maximal activity
at 50°C and pH 9~10, in 20% NaCl and was highly stabilized by
NaCl. It was able to hydrolyse natural actomyosin (NAM), collagen
and anchovy protein. For NAM hydrolysis, the myosin heavy chain
was completely digested by halophilic proteinase as evidenced by the
lowest band intensity remaining, but partially hydrolysed actin. The
SR5-3 proteinase was also capable hydrolyzing two major
components of collagen, β- and α-compounds, effectively. The
degree of hydrolysis (DH) of the halophilic proteinase and
commercial proteinases (Novozyme, Neutrase, chymotrypsin and
Flavourzyme) on the anchovy protein, were compared, and it was
found that the proteinase showed a greater degree of hydrolysis
towards anchovy protein than that from commercial proteinases. DH
of halophilic proteinase was sharply enhanced according to the
increase in the concentration of enzyme from 0.035 U to 0.105 U.
The results warranting that the acceleration of the production of fish
sauce with higher quality, may be achieved by adding of the
halophilic proteinase from this bacterium.
[1] K. Lopetcharat, J. W. Park, "Characteristics of fish sauce made from
Pacific whiting and surimi by-products during fermentation storage," J.
Food Sci, vol. 67, pp. 511-516, 2002.
[2] P. Saisithi, B. Kasemsarn, J. J. Liston, A. M. Dollar, "Microbiology and
chemistry of fermented fish," J. Food Sci, vol. 31, pp. 105-110, 1996.
[3] F. M. Orejana, J. Liston, "Agents of proteolysis and its inhibition in
Patis (fish sauce) fermentation," J. Food Sci, vol. 47, pp. 198-203. 1981.
[4] B. K. Simpson, "Digestive proteinases from marine animals". In N. F.
Haard and B. K. Simpson (Eds.), Seafood enzymes: Utilization and
influence on postharvest seafood quality. 531-540. New York, USA:
Mercel Dekker. 2000.
[5] S. Klomklao, S. Benjakul, W. Visessanguan, "Comparative studies on
proteolytic activity of spleen extracts from three tuna species commonly
used in Thailand," J. Food Biochem, vol. 28, pp. 355-372, 2004.
[6] S. Klomklao, S. Benjakul, W. Visessanguan, H. Kishimura, B. K.
Simpson, "Proteolytic degradation of sardine (Sardinella gibbosa)
proteins by trypsin from skipjack tuna (Katsuwonus pelamis) spleen,"
Food Chem, vol. 98, pp. 14-22, 2006.
[7] K. Hiraga, Y. Nishikata, S. Namwong, S. Tanasupawat, T. Takada, K.
Oda, "Purification and characterization of serine proteinase from
halophilic bacterium, Filobacillus sp. RF2-5," Biosci. Biotechnol.
Biochem, vol. 69, pp. 38-44, 2005.
[8] S. Namwong, K. Hiraga, K. Takada, S. Tanasupawat, K. Oda, "A
halophilic serine proteinase from Halobacillus sp. SR5-3 isolated from
fish sauce: purification and characterization," Biosci. Biotechnol.
Biochem, vol. 70(6), pp. 1395-1401, 2006.
[9] R. Chaveesuk, J. P. Smith, B. K. Simpson, "Production of fish sauce and
acceleration of sauce fermentation using proteolytic enzymes," J. Aquat.
Food Prod. Tech, vol. 2(3), pp. 59-77, 1993.
[10] C. Sanchez-Porro, E, Mellado, C. Bertoldo, G. Antranikian, A. Ventosa,
"Screening and characterization of the proteinase CP1 produced by the
moderately halophilic bacterium Pseudomonas sp. strain CP76,"
Extremophiles, vol. 7, pp. 221-228, 2003.
[11] H. Capiralla, T. Hiroi, T. Hirokawa, S. Maeda, "Purification and
characterization of a hydrophobic amino acid-specific endopeptidase
form Halobacterium halobium S9 with potential application in
debittering of protein hydrolysates," Process Biochem, vol. 38, pp. 571-
579, 2002.
[12] O. H. Lowry, N. J. Rosebrough, A. L. Farr, R. J. Randall, "Protein
measurement with the folin phenol reagent," J. Biol. Chem, vol. 193, pp.
265-275, 1951.
[13] S. Benjakul, T. A. Seymour, M. T. Morrissey, H. An,(1997).
"Physicochemical changes in Pacific whiting muscle proteins during ice
storage," J. Food Sci, vol. 62, pp. 729-733, 1997.
[14] S. Benjakul, W. Visessanguan, K. Leelapongwattana, "Purification and
characterization of heat-stable alkaline proteinase from bigeye snapper
(Priacanthus macracanthus) muscle," Comp. Biochem. Phys. B, vol. 24,
pp. 107-127, 2003.
[15] S. Benjakul, M. Morrissey, "Protein hydrolysates from Pacific whiting
solid wastes," J. Agri. Food Chem, vol. 45, pp. 3423-3430, 1995.
[16] U. K. Laemmli, "Cleavage of structure proteins during the assembly of
the head of bacteriophage T4," Nature, vol. 277, pp. 680-685, 1970.
[17] M. J. Cao, K. Osatomi, K. Hara,T. Ishihara, "Identification of a
myofibril-bound serine proteinase (MBSP) in the skeletal muscle of
lizard fish Saurida wanieso which specifically cleaves the arginine site,"
Comp. Biochem. Phys. B, vol. 125, pp. 255-264, 2000.
[18] K. Osatomi, H. Sasai, M. Cao, K. Hara, T. Ishihara, "Purification and
characterization of myofibril-bound serine proteinase from carp,
Cyprinus carpio, ordinary muscle," Comp. Biochem. Phys. B, vol. 16,
pp. 183-190, 1997.
[19] T. Aoki, R. Ueno, "Involvement of cathepsins B and L in the postmortem
autolysis of mackerel muscle" Food Res. Int, vol. 30(8), pp.
585-591, 1997.
[20] K. Ryu, J. Kim, J. S. Dordick, "Catalytic properties and potential of an
extracellular protease from an extreme halophile," Enzyme Microbiol.
Tech, vol. 16(4), pp. 266-275, 1994.
[21] V. M. Stepanov, G. N. Rudenskaya, L. P. Revina, Y. B. Gryaznova, E.
N. Lysogorskaya, I. Y. Filippova, I. I. Ivanova, "A serine protease of an
archaebacterium, Halobacterium maditerranei," Biochem. J, vol. 285,
pp. 281-286, 1992.
[22] L. S. Izotova, A. Y. Strongin, L. N. Chekulaeva, V. E. Sterkin, V. I.
Ostoslavskaya, L. A. Lyublinskaya, E. A. Timokhina, V. M. Stepanov,
"Purification and properties of serine protease from Halobacterium
halobium," J. Bacteriol, vol. 155, pp. 826-830, 1983.
[23] M. Kamekura, Y. Seno, "Partial sequence of the gene for a serine
protease from a halophilic archaeum Haloferax mediterranei R4, and
nucleotide sequence of 16S rRNA encoding genes from several
halophilic archaea," Experimentia, vol. 49, pp. 503-513, 1993.
[24] S. C. Hathwar, B. Bijinu, A. K. Rai, B. Narayan, "Simultaneous recovery
of lipids and proteins by enzymatic hydrolysis of fish industry waste
using different commercial proteases," Appl. Biochem. Biotechnol, vol.
164, pp. 115-124, 2011.
[1] K. Lopetcharat, J. W. Park, "Characteristics of fish sauce made from
Pacific whiting and surimi by-products during fermentation storage," J.
Food Sci, vol. 67, pp. 511-516, 2002.
[2] P. Saisithi, B. Kasemsarn, J. J. Liston, A. M. Dollar, "Microbiology and
chemistry of fermented fish," J. Food Sci, vol. 31, pp. 105-110, 1996.
[3] F. M. Orejana, J. Liston, "Agents of proteolysis and its inhibition in
Patis (fish sauce) fermentation," J. Food Sci, vol. 47, pp. 198-203. 1981.
[4] B. K. Simpson, "Digestive proteinases from marine animals". In N. F.
Haard and B. K. Simpson (Eds.), Seafood enzymes: Utilization and
influence on postharvest seafood quality. 531-540. New York, USA:
Mercel Dekker. 2000.
[5] S. Klomklao, S. Benjakul, W. Visessanguan, "Comparative studies on
proteolytic activity of spleen extracts from three tuna species commonly
used in Thailand," J. Food Biochem, vol. 28, pp. 355-372, 2004.
[6] S. Klomklao, S. Benjakul, W. Visessanguan, H. Kishimura, B. K.
Simpson, "Proteolytic degradation of sardine (Sardinella gibbosa)
proteins by trypsin from skipjack tuna (Katsuwonus pelamis) spleen,"
Food Chem, vol. 98, pp. 14-22, 2006.
[7] K. Hiraga, Y. Nishikata, S. Namwong, S. Tanasupawat, T. Takada, K.
Oda, "Purification and characterization of serine proteinase from
halophilic bacterium, Filobacillus sp. RF2-5," Biosci. Biotechnol.
Biochem, vol. 69, pp. 38-44, 2005.
[8] S. Namwong, K. Hiraga, K. Takada, S. Tanasupawat, K. Oda, "A
halophilic serine proteinase from Halobacillus sp. SR5-3 isolated from
fish sauce: purification and characterization," Biosci. Biotechnol.
Biochem, vol. 70(6), pp. 1395-1401, 2006.
[9] R. Chaveesuk, J. P. Smith, B. K. Simpson, "Production of fish sauce and
acceleration of sauce fermentation using proteolytic enzymes," J. Aquat.
Food Prod. Tech, vol. 2(3), pp. 59-77, 1993.
[10] C. Sanchez-Porro, E, Mellado, C. Bertoldo, G. Antranikian, A. Ventosa,
"Screening and characterization of the proteinase CP1 produced by the
moderately halophilic bacterium Pseudomonas sp. strain CP76,"
Extremophiles, vol. 7, pp. 221-228, 2003.
[11] H. Capiralla, T. Hiroi, T. Hirokawa, S. Maeda, "Purification and
characterization of a hydrophobic amino acid-specific endopeptidase
form Halobacterium halobium S9 with potential application in
debittering of protein hydrolysates," Process Biochem, vol. 38, pp. 571-
579, 2002.
[12] O. H. Lowry, N. J. Rosebrough, A. L. Farr, R. J. Randall, "Protein
measurement with the folin phenol reagent," J. Biol. Chem, vol. 193, pp.
265-275, 1951.
[13] S. Benjakul, T. A. Seymour, M. T. Morrissey, H. An,(1997).
"Physicochemical changes in Pacific whiting muscle proteins during ice
storage," J. Food Sci, vol. 62, pp. 729-733, 1997.
[14] S. Benjakul, W. Visessanguan, K. Leelapongwattana, "Purification and
characterization of heat-stable alkaline proteinase from bigeye snapper
(Priacanthus macracanthus) muscle," Comp. Biochem. Phys. B, vol. 24,
pp. 107-127, 2003.
[15] S. Benjakul, M. Morrissey, "Protein hydrolysates from Pacific whiting
solid wastes," J. Agri. Food Chem, vol. 45, pp. 3423-3430, 1995.
[16] U. K. Laemmli, "Cleavage of structure proteins during the assembly of
the head of bacteriophage T4," Nature, vol. 277, pp. 680-685, 1970.
[17] M. J. Cao, K. Osatomi, K. Hara,T. Ishihara, "Identification of a
myofibril-bound serine proteinase (MBSP) in the skeletal muscle of
lizard fish Saurida wanieso which specifically cleaves the arginine site,"
Comp. Biochem. Phys. B, vol. 125, pp. 255-264, 2000.
[18] K. Osatomi, H. Sasai, M. Cao, K. Hara, T. Ishihara, "Purification and
characterization of myofibril-bound serine proteinase from carp,
Cyprinus carpio, ordinary muscle," Comp. Biochem. Phys. B, vol. 16,
pp. 183-190, 1997.
[19] T. Aoki, R. Ueno, "Involvement of cathepsins B and L in the postmortem
autolysis of mackerel muscle" Food Res. Int, vol. 30(8), pp.
585-591, 1997.
[20] K. Ryu, J. Kim, J. S. Dordick, "Catalytic properties and potential of an
extracellular protease from an extreme halophile," Enzyme Microbiol.
Tech, vol. 16(4), pp. 266-275, 1994.
[21] V. M. Stepanov, G. N. Rudenskaya, L. P. Revina, Y. B. Gryaznova, E.
N. Lysogorskaya, I. Y. Filippova, I. I. Ivanova, "A serine protease of an
archaebacterium, Halobacterium maditerranei," Biochem. J, vol. 285,
pp. 281-286, 1992.
[22] L. S. Izotova, A. Y. Strongin, L. N. Chekulaeva, V. E. Sterkin, V. I.
Ostoslavskaya, L. A. Lyublinskaya, E. A. Timokhina, V. M. Stepanov,
"Purification and properties of serine protease from Halobacterium
halobium," J. Bacteriol, vol. 155, pp. 826-830, 1983.
[23] M. Kamekura, Y. Seno, "Partial sequence of the gene for a serine
protease from a halophilic archaeum Haloferax mediterranei R4, and
nucleotide sequence of 16S rRNA encoding genes from several
halophilic archaea," Experimentia, vol. 49, pp. 503-513, 1993.
[24] S. C. Hathwar, B. Bijinu, A. K. Rai, B. Narayan, "Simultaneous recovery
of lipids and proteins by enzymatic hydrolysis of fish industry waste
using different commercial proteases," Appl. Biochem. Biotechnol, vol.
164, pp. 115-124, 2011.
@article{"International Journal of Biological, Life and Agricultural Sciences:57528", author = "Sirilak Namwong and Wonnop Visessanguan and Soottawat Benjakul and Somboon Tanasupawat", title = "Proteolytic Dedradation of Anchovy (Spolephorus spp.) Proteins by Halophilic Proteinase from Halobacillus sp. SR5-3", abstract = "The halophilic proteinase showed a maximal activity
at 50°C and pH 9~10, in 20% NaCl and was highly stabilized by
NaCl. It was able to hydrolyse natural actomyosin (NAM), collagen
and anchovy protein. For NAM hydrolysis, the myosin heavy chain
was completely digested by halophilic proteinase as evidenced by the
lowest band intensity remaining, but partially hydrolysed actin. The
SR5-3 proteinase was also capable hydrolyzing two major
components of collagen, β- and α-compounds, effectively. The
degree of hydrolysis (DH) of the halophilic proteinase and
commercial proteinases (Novozyme, Neutrase, chymotrypsin and
Flavourzyme) on the anchovy protein, were compared, and it was
found that the proteinase showed a greater degree of hydrolysis
towards anchovy protein than that from commercial proteinases. DH
of halophilic proteinase was sharply enhanced according to the
increase in the concentration of enzyme from 0.035 U to 0.105 U.
The results warranting that the acceleration of the production of fish
sauce with higher quality, may be achieved by adding of the
halophilic proteinase from this bacterium.", keywords = "Halophilic proteinase, Halobacillus sp. SR5-3,
anchovy (Spolephorus spp.) proteins, fish sauce", volume = "6", number = "8", pages = "603-5", }
