Optimization of cultural conditions for protease production by a fungal species

P. Kamath, V.M. Subrahmanyam, J. Rao, P. Raj

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Studies were carried out on a paddy soil fungal isolate identified to be a strain of Aspergillus niger from Manipal. The parameters that largely impact enzyme production viz., fermentation time, impeller speed, pH, temperature and nutrient supplements were studied. Optimization of production parameters for production of protease was done by the single-parameter mode. Casein served as substrate and proteolytic activity was estimated using Folin-Ciocalteau method at 660 nm. A maximum yield of 71.3 mg tyrosine/g casein substrate was produced in 96 h on a soluble starch medium at pH 4 in shake flask experiments. Production was carried out on a 3-liter fermenter and 40.7 mg of tyrosine was liberated/g of substrate. The enzyme was extracted with 50% ammonium sulfate and sodium dodecyl sulfate-Polyacrylamide gel electrophoresis showed two bands having mw 45.7 kDa and 38.5 kDa, respectively. The enzyme activity was found to be 147.84 U/ml.
Original languageEnglish
Pages (from-to)161-166
Number of pages6
JournalIndian Journal of Pharmaceutical Sciences
Volume72
Issue number2
DOIs
Publication statusPublished - 2010

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Peptide Hydrolases
Caseins
Tyrosine
Enzymes
Aspergillus niger
Ammonium Sulfate
Sodium Dodecyl Sulfate
Starch
Fermentation
Polyacrylamide Gel Electrophoresis
Soil
Food
Temperature
folin

Cite this

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title = "Optimization of cultural conditions for protease production by a fungal species",
abstract = "Studies were carried out on a paddy soil fungal isolate identified to be a strain of Aspergillus niger from Manipal. The parameters that largely impact enzyme production viz., fermentation time, impeller speed, pH, temperature and nutrient supplements were studied. Optimization of production parameters for production of protease was done by the single-parameter mode. Casein served as substrate and proteolytic activity was estimated using Folin-Ciocalteau method at 660 nm. A maximum yield of 71.3 mg tyrosine/g casein substrate was produced in 96 h on a soluble starch medium at pH 4 in shake flask experiments. Production was carried out on a 3-liter fermenter and 40.7 mg of tyrosine was liberated/g of substrate. The enzyme was extracted with 50{\%} ammonium sulfate and sodium dodecyl sulfate-Polyacrylamide gel electrophoresis showed two bands having mw 45.7 kDa and 38.5 kDa, respectively. The enzyme activity was found to be 147.84 U/ml.",
author = "P. Kamath and V.M. Subrahmanyam and J. Rao and P. Raj",
note = "Cited By :16 Export Date: 10 November 2017 CODEN: IJSID Correspondence Address: Subrahmanyam, V. M.; Department of Pharmaceutical Biotechnology, Manipal College of Pharmaceutical Sciences, Manipal University, Manipal-576 104, India; email: vm.subra@manipal.edu Chemicals/CAS: ammonium sulfate, 7783-20-2; casein, 9000-71-9; dodecyl sulfate sodium, 151-21-3; proteinase, 9001-92-7 References: Negi, S., Banerjee, R., Optimization of amylase and protease production from Aspergillus awamori in single bioreactor through EVOP factorial design technique (2006) Food Technol Biotechnol, 44, pp. 257-261; Yadwad, V.B., Wilson, S., Ward, O.P., Effect of culture conditions and induction strategies on production of human interleukin-6 by a recombinant Aspergillus nidulans strain (1996) Mycol Res, 100, pp. 356-360; Chandran, S., Alagarsamy, S., Pandey, A., Webb, C., Soccol, C.R., Larroche, C., Proteases in enzyme technology (2004) Enzyme Technology Vol. XVIII, pp. 312-325. , Pandey A, Webb C, Soccol CR, Larroche C. Editors 1st edition, New Delhi, Asiatech Publishers Inc; Dai, Z., Mao, X., Magnuson, J.K., Lasure, L.L., Identification of Genes associated with morphology in Aspergillus niger by using suppression subtractive hybridization (2003) Appl Environ Microbiol, 70, pp. 2474-2485; Folin, O., Ciocalteau, V., Tyrosine and Tryptophan determinations in proteins (1927) J Biol Chem, 73, pp. 627-650; Olajuyigbe, F.M., Ajele, J.O., Olawoye, T.L., Soe physicochemical properties of acid protease produced during growth of Aspergillus niger (NRRL 1785) (2003) Global J Pure Appl Sci, 9, pp. 523-528; Samarntarn, W., Cheevadhanarak, S., Tanticharoen, M., Production of alkaline protease by a genetically engineered Aspergillus oryzae U1521 (1999) J Gen Appl Microbiol, (45), pp. 99-103; O'Donnell, D., Xua, W.L., Ridgway, D.J., Gua, T., Moo-Young, M., Enhanced heterologous protein production in Aspergillus niger through pH control of extracellular protease activity (2001) Biochem Eng J, 8, pp. 187-193; Rahman, R.N.Z.R.A., Geok, L.P., Basri, M., Salleh, A.B., An organic solvent-tolerant protease from Pseudomonas aeruginosa strain K: Nutritional factors affecting protease production (2005) Enzyme and Microbial Technology, 36 (5-6), pp. 749-757. , DOI 10.1016/j.enzmictec.2004.12.022, PII S0141022904004065; Shumi, W., Hussain Md, T., Anwar, M.N., Isolation and purification of fungus Aspergillus funiculosus and its enzyme protease (2004) Pak J Biol Sci, 7, pp. 312-317; Dahot, M.U., Purification and some properties of alkaline protease from Penicillum expansum (1994) J Islam Acad Sci, 7, p. 2; Patke, D., Dey, S., Proteolytic activity from a thermophilic Streptomyces megasporus strain SDP4 (1998) Lett Appl Microbiol, 26, pp. 171-174; De Azeredo, L.A., De Lima, M.B., Coelho, R.R., Freire, D.M., Thermophilic protease production by Streptomyces sp. 594 in submerged and solid-state fermentations using feather meal (2006) J Appl Microbiol, 100, pp. 641-647",
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volume = "72",
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Optimization of cultural conditions for protease production by a fungal species. / Kamath, P.; Subrahmanyam, V.M.; Rao, J.; Raj, P.

In: Indian Journal of Pharmaceutical Sciences, Vol. 72, No. 2, 2010, p. 161-166.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Optimization of cultural conditions for protease production by a fungal species

AU - Kamath, P.

AU - Subrahmanyam, V.M.

AU - Rao, J.

AU - Raj, P.

N1 - Cited By :16 Export Date: 10 November 2017 CODEN: IJSID Correspondence Address: Subrahmanyam, V. M.; Department of Pharmaceutical Biotechnology, Manipal College of Pharmaceutical Sciences, Manipal University, Manipal-576 104, India; email: vm.subra@manipal.edu Chemicals/CAS: ammonium sulfate, 7783-20-2; casein, 9000-71-9; dodecyl sulfate sodium, 151-21-3; proteinase, 9001-92-7 References: Negi, S., Banerjee, R., Optimization of amylase and protease production from Aspergillus awamori in single bioreactor through EVOP factorial design technique (2006) Food Technol Biotechnol, 44, pp. 257-261; Yadwad, V.B., Wilson, S., Ward, O.P., Effect of culture conditions and induction strategies on production of human interleukin-6 by a recombinant Aspergillus nidulans strain (1996) Mycol Res, 100, pp. 356-360; Chandran, S., Alagarsamy, S., Pandey, A., Webb, C., Soccol, C.R., Larroche, C., Proteases in enzyme technology (2004) Enzyme Technology Vol. XVIII, pp. 312-325. , Pandey A, Webb C, Soccol CR, Larroche C. Editors 1st edition, New Delhi, Asiatech Publishers Inc; Dai, Z., Mao, X., Magnuson, J.K., Lasure, L.L., Identification of Genes associated with morphology in Aspergillus niger by using suppression subtractive hybridization (2003) Appl Environ Microbiol, 70, pp. 2474-2485; Folin, O., Ciocalteau, V., Tyrosine and Tryptophan determinations in proteins (1927) J Biol Chem, 73, pp. 627-650; Olajuyigbe, F.M., Ajele, J.O., Olawoye, T.L., Soe physicochemical properties of acid protease produced during growth of Aspergillus niger (NRRL 1785) (2003) Global J Pure Appl Sci, 9, pp. 523-528; Samarntarn, W., Cheevadhanarak, S., Tanticharoen, M., Production of alkaline protease by a genetically engineered Aspergillus oryzae U1521 (1999) J Gen Appl Microbiol, (45), pp. 99-103; O'Donnell, D., Xua, W.L., Ridgway, D.J., Gua, T., Moo-Young, M., Enhanced heterologous protein production in Aspergillus niger through pH control of extracellular protease activity (2001) Biochem Eng J, 8, pp. 187-193; Rahman, R.N.Z.R.A., Geok, L.P., Basri, M., Salleh, A.B., An organic solvent-tolerant protease from Pseudomonas aeruginosa strain K: Nutritional factors affecting protease production (2005) Enzyme and Microbial Technology, 36 (5-6), pp. 749-757. , DOI 10.1016/j.enzmictec.2004.12.022, PII S0141022904004065; Shumi, W., Hussain Md, T., Anwar, M.N., Isolation and purification of fungus Aspergillus funiculosus and its enzyme protease (2004) Pak J Biol Sci, 7, pp. 312-317; Dahot, M.U., Purification and some properties of alkaline protease from Penicillum expansum (1994) J Islam Acad Sci, 7, p. 2; Patke, D., Dey, S., Proteolytic activity from a thermophilic Streptomyces megasporus strain SDP4 (1998) Lett Appl Microbiol, 26, pp. 171-174; De Azeredo, L.A., De Lima, M.B., Coelho, R.R., Freire, D.M., Thermophilic protease production by Streptomyces sp. 594 in submerged and solid-state fermentations using feather meal (2006) J Appl Microbiol, 100, pp. 641-647

PY - 2010

Y1 - 2010

N2 - Studies were carried out on a paddy soil fungal isolate identified to be a strain of Aspergillus niger from Manipal. The parameters that largely impact enzyme production viz., fermentation time, impeller speed, pH, temperature and nutrient supplements were studied. Optimization of production parameters for production of protease was done by the single-parameter mode. Casein served as substrate and proteolytic activity was estimated using Folin-Ciocalteau method at 660 nm. A maximum yield of 71.3 mg tyrosine/g casein substrate was produced in 96 h on a soluble starch medium at pH 4 in shake flask experiments. Production was carried out on a 3-liter fermenter and 40.7 mg of tyrosine was liberated/g of substrate. The enzyme was extracted with 50% ammonium sulfate and sodium dodecyl sulfate-Polyacrylamide gel electrophoresis showed two bands having mw 45.7 kDa and 38.5 kDa, respectively. The enzyme activity was found to be 147.84 U/ml.

AB - Studies were carried out on a paddy soil fungal isolate identified to be a strain of Aspergillus niger from Manipal. The parameters that largely impact enzyme production viz., fermentation time, impeller speed, pH, temperature and nutrient supplements were studied. Optimization of production parameters for production of protease was done by the single-parameter mode. Casein served as substrate and proteolytic activity was estimated using Folin-Ciocalteau method at 660 nm. A maximum yield of 71.3 mg tyrosine/g casein substrate was produced in 96 h on a soluble starch medium at pH 4 in shake flask experiments. Production was carried out on a 3-liter fermenter and 40.7 mg of tyrosine was liberated/g of substrate. The enzyme was extracted with 50% ammonium sulfate and sodium dodecyl sulfate-Polyacrylamide gel electrophoresis showed two bands having mw 45.7 kDa and 38.5 kDa, respectively. The enzyme activity was found to be 147.84 U/ml.

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DO - 10.4103/0250-474X.65017

M3 - Article

VL - 72

SP - 161

EP - 166

JO - Indian Journal of Pharmaceutical Sciences

JF - Indian Journal of Pharmaceutical Sciences

SN - 0250-474X

IS - 2

ER -