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Research Article
 

Purification and Some Properties of Acid Protease from Penicillium expansum



M. Umar Dahot
 
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ABSTRACT

An acid protease was isolated from the culture broth of Penicillium expansum grown on rice husk (40 mesh fine powder) mineral medium. The protease enzyme was purified on Sephadex G-100 and DEAE Sephdex A-50 with recovery of 7.52%. Purified enzyme shows a single band on SDS polyacrylamide gel electrophoresis. The optimum activity of protease was found at pH 3.5 and temperature 30ºC. The activation energy for the hydrolysis of casien by acid protease was 93 KJ/mol. The enzyme activity was highly increased in the presence of CoCl2 (71.57%), cysteine (54.23%), mercaptoethanol (44.70%), ZnCl2 (34.78%) but slightly activated by CaCl2 (10.24%) and MnCl2 (1.37%). Whereas protease activity was reduced on the addition of EDTA (55.88%), AgNo3 (67.85%) and HgNO3)2 (64.71%). The thermal stability of protease was increased in the presence of ZnCl2. The half-life of acid protease in the absence and presence of ZnCl2 at 50ºC was determined 17 and 28 minutes respectively.

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  How to cite this article:

M. Umar Dahot , 2001. Purification and Some Properties of Acid Protease from Penicillium expansum. Journal of Applied Sciences, 1: 405-408.

DOI: 10.3923/jas.2001.405.408

URL: https://scialert.net/abstract/?doi=jas.2001.405.408

REFERENCES
Arvidsona, S., T. Holmea and B. Lindholm, 1973. Studies on extracellular proteolytic enzymes from Staphylococcus aureus: I. Purification and characterization of one neutral and one alkaline protease. Biochimica et Biophysica Acta (BBA)-Enzymol., 302: 135-148.
CrossRef  |  

Burrell, R.G., C.W. Clayton, M.E. Gallegly and V.G. Lilley, 1966. Factors affecting the antigenicity of the mycelium of three species of phytophthora. Phytopathology, 56: 422-426.

Cowan, D.A., K.A. Smolenski, R.M. Daniel and H.W. Morgan, 1987. An extremely thermostable extracellular proteinase from a strain of the archaebacterium Desulfurococcus growing at 88°C. Biochem. J., 247: 121-133.
Direct Link  |  

Dahot, M.U., 1987. Studies on proteolytic enzyme Part 1. Characteristics of protease synthesis by Penicillium expansum. Pak. J. Sci. Ind. Res., 30: 194-196.
Direct Link  |  

Dahot, M.U., 1993. Cultivation of Penicillium expansum on rice husk powder for protease production. J. Islamic Acad. Sci., 6: 193-196.
Direct Link  |  

Dahot, M.U., 1994. Purification and some properties of alkaline protease from Pensillium expansum. J. Islamic Acad. Sci., 7: 100-105.
Direct Link  |  

Dahot, M.U., 1995. Purification and characterization of an extracellular protease produced by Penicillium expansum. J. I. R. Iran, 6: 131-135.

Davis, B.J., 1964. Disc electrophoresis-II, method and application to human serum proteins. Ann. N. Y. Acad. Sci., 121: 404-427.
CrossRef  |  PubMed  |  Direct Link  |  

Eriksson, K.E. and B. Pettersson, 1982. Purification and partial characterization of two acidic proteases from the white-rot fungus Sporotrichum pulverulentum. Eur. J. Biochem. 124: 635-642.
Direct Link  |  

Hashimoto, H., Y. Kaneko, T. Iwaasa and T. Yokotsuka, 1973. Production and purification of acid protease from thermophilic fungus Penicillium duponti K1014. Applied Microbiol., 25: 584-588.
Direct Link  |  

Ichishima, E., 1970. Aspartic Proteinases of A Saitol. In: Methods in Enzymology, Perlmann, G.E. and L. Lorand (Eds.). Academic Press, New York, pp: 397.

Lowry, O.H., N.J. Rosebrough, A.L. Farr and R.J. Randall, 1951. Protein measurement with the folin phenol reagent. J. Biol. Chem., 193: 265-275.
PubMed  |  Direct Link  |  

Majima, E., K. Oda, S. Murao and E. Ichishima, 1988. Comparative study on the specificities of several fungal aspartic and acidic proteinases towards the tetradecapeptide of a renin substrate. Agric. Biol. Chem., 52: 787-793.
Direct Link  |  

McLaughlin, J. and G. Faubert, 1977. Partial purification and some properties of a neutral sulfhydryl and an acid proteinase from Entamoeba histolytica. Can. J. Microbiol., 23: 420-425.
Direct Link  |  

Murakami, K., Y. Ishida, A. Masaki, H. Tatsumi and S. Murakami et al., 1991. Isolation and characterization of the alkaline protease gene of Aspergillus oryzae. Agric. Biol. Chem., 55: 2807-2811.
Direct Link  |  

Oda, K., H. Torishima and S. Murao, 1986. Purification and characterization of acid proteinase C of Scytalidium hgnicolum ATCC 24568. Agric. Biol. Chem., 50: 651-658.
Direct Link  |  

Oda, K., T. Nakuzima, T. Terashita, K. Suzuki and S. Murao, 1987. Purification and properties of an S-PI (Pepstatin Ac)-insensitive carboxyl proteinase from a Xanthomonas sp. bacterium. Agric. Biol. Chem., 51: 3073-3080.
Direct Link  |  

Takeuchi, M., T. Ushijima and E. Ichishima, 1986. Acid carboxypeptidase from Aspergillus oryzae. Agric. Biol. Chem., 50: 633-638.
Direct Link  |  

Toogood, H.S., M. Prescott and R.M. Daniel, 1995. A pepstatin-insensitive aspartic proteinase from a thermophilic Bacillus sp. Biochem. J., 307: 783-789.
Direct Link  |  

Tsujita, Y. and A. Endo, 1977. Extracellular acid protease of Aspergillus oryzae grown on liquid media: Multiple forms due to association with heterogeneous polysaccharides. J. Bacteriol., 130: 48-56.
Direct Link  |  

Vasantha, N., L.D. Thompson, C. Rhodes, C. Banner, J. Nagle and D. Filpula, 1984. Genes for alkaline protease and neutral protease from Bacillus amyloliquefaciens contain a large open reading frame between the regions coding for signal sequence and mature protein. J. Bacteriol., 159: 811-819.
Direct Link  |  

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