Partial characterization of proteases from Citrus sinensis fruit peel
Languages of publication
Proteases are one of the most important enzymes that have various physiological and industrial applications. This study was carried out to purify and partially characterize proteases from Citrus sinensis fruit peel. Three active fractions of the proteases (I, II and III) were obtained. The Vmax for proteases I, II, III and pooled fraction were 185.19, 192.31, 111.11 and 163.93 U/ml with Michaelis-Menten’s constant (Km) 1.01, 0.44, 0.67 and 0.37 mg/ml respectively. The enzymes were optimally active at 40-50 °C. However, they retained activity at 60-70 °C. Protease I was stable up to 60 °C while proteases II and III retained more than 80% activity in the range of 25-70 °C. The optimal pH of proteases II and III was 7 while protease I was optimally active at pH 8. The enzymes were stable at alkaline pH especially between 6 and 9 retaining more than 60% of its activity. The stability of these enzymes at high temperature and different pH may be an indication of its potential applications in food, chemical and laundry industries.
- Organic and Natural Products Division, Basic Research Department, National Research Institute for Chemical Technology, Zaria, Kaduna State, Nigeria, firstname.lastname@example.org
- Department of Biochemistry, Faculty of Life Sciences, University of Ilorin, Ilorin, Nigeria
-  Abusham, R.A., Rahman, R.N.Z.A., Salleh, A.B. and Basri, M. (2009). Optimization of physical factors affecting the production of thermo-stable organic solvent-tolerant protease from a newly isolated halotolerant Bacillus subtilis strain Rand. Microbiology and Cell Facts 8; 20.
-  Adinarayana, K., Ellaiah, P. and Prasad, S.D. (2003). Purification and partial characterization of thermostable serine alkaline protease from a newly isolated Bacillus subtilis PE-11. AAPS Pharmaceutical Science and Technology 4; 1-9.
-  Antao, C.M. and Malcata, F.X. (2005). Plant serine proteases: biochemical, physiological and molecular features. Plant Physiology and Biochemistry 43; 637-650.
-  Benito, M.I., Rodriguez, M., Núñez, F., Asensio, M.A., Bermúdez, M.E. and Córdoba, J.J. (2002). Purification and characterization of an extracellular protease from Penicillium chrsogenun Pg222 active against meat proteins. Applied Environmental Microbiology 68; 3532-3536.
-  Berger, D. and Altmann, I. (2000). A subtilisin-like serine protease involved in the regulation of stomatal density and distribution in Arabidopsi thaliana. Genes Development 14; 1119-1131.
-  Bushra, M., Ahsan, H., Abdul, M. and Hassan, Z. (2010). Purification and characterization of alkaline protease from Aspergillus terreus. Journal of Chemical Society of Pakistan 32; 497-503.
-  Chen, K.Q., Robinson, A.C., Van Dam, M.E., Martinez, P., Economou, C. and Arnold, F.H. (1991). Enzyme engineering for nonaqueous solvents, II. Additive effects of mutations on the solubility and activity of subtilisin E in polar organic media. Biotechnology Programme. 7; 125-129.
-  Chen, T.E., Huang, D.J. and Lin, Y.H. (2004). Isolation and characterization of a serine protease from the storage roots of sweet potato (Ipomoea batatas [L.] Lam). Plant Science 166; 1019–1026.
-  Devi, M.K., Banu, A.R., Gnanaprabhal, G.R., Pradeep, B.V. and Palaniswamy, M. (2008). Purification, characterization of alkaline protease enzyme from native isolate Aspergillus niger and its compatibility with commercial detergents. Indian Journal of Science and Technology 1; 1-6.
-  Ekpa, E., Nzelibe, H.C. and Onyike, E. (2010). Isolation of cysteine protease from banana agrowastes. Nigerian Journal of Biochemistry and Molecular Biology 25(3); 39-45.
-  Evans, C., Egwim, J. Yisa and P.O. Egwim (2009). Kinetics studies of protease in fermenting locust beans (Parkia biglobosa) and melon seed (Citrullus vulgaris). African Journal of Biochemical Research 3(4); 145-149.
-  Fahmy, A.S. and Fahmy, S.A. (2003). A serine protease from germinated wheat seeds. Plant Biology (Rockville). 38.
-  Fahmy, A.S., Ali, A.A. and Mohamed, S.A. (2004). Characterization of a cysteine protease from wheat Triticum aestivum (cv. Giza 164), Biology and Technology 91; 297-304.
-  Gaur, S. and Wadhwa, P.A. (2008). Alkaline proteases from seneseed leaves of invasive weed Lantana camara. African Journal of Biotechnology 12; 134-141.
-  Germano, S., Pandy, A., Osaku, C.A., Rocha, S.N. and Soccol, C.R. (2003). Characterization and stability of protease from Penicillium sp. Produced by solid-state fermentation. Enzyme Microbiology and Technology 32; 246-251.
-  Gupta, R., Beg, Q.K. and Lorenz, P. (2002). Bacterial alkaline protease: molecular approaches and industrial application. Applied Microbial Biotechnology 59; 15-32.
-  Hamilton, J.M.U., Simpson, D.J., Hyman, S.C., Ndimba, B.G. and Slabas, A.R. (2003). Ara12 subtilisin-like protease from Arabidopsis thaliana: purification, substrate specificity and tissue localization. Journal of Biochemistry 370; 57-67.
-  Ire, S.F., Bartholomew, N.O. and Anene, A.M. (2011). Purification and characterization of an acid protease from Aspergillus carbonarius. African Journal of Food Science 5(12); 695-709.
-  Jaouadi, B., Ellouz-Chaabouni, S., Rhimi, M. and Bejar, S. (2008). Biochemical and molecular characterization of a detergent stable serine alkaline protease from Bacillus pumilus CBS with high catalytic efficiency. Biochimie 90; 1291-1305.
-  Jones, B.L. (2005). Endoproteases of barley and malt. Journal of Cereal Sciences 42; 139-156.
-  Joo, H.S., Kumar, C.G., Park, G.C., Paik, S.R. and Chang, C.S. (2003). Oxidant and SDS stable alkaline protease from Bacillus clausii I-52: production and some properties. Journal of Applied Microbiology 95; 267-272.
-  Khan, H., Subhan, M., Mehmood, S., Durrani, F. M., Abbas, S. and Khan, S. (2008). Purification and characterization of serine protease from seeds of Holarrhena antidysenterica. Biotechnology 7 (1); 94-99.
-  Leary, D., Vierros, M., Hamon, G., Arico, S. and Monagle, C. (2009). Marine genetic resources: a review of scientific and commercial interest. Marine Policy 33; 183-194.
-  Mohamed, S.A. (2005). Purification and characterization of a serine protease from wheat. FEBS Journal 272; 171-171.
-  Ndidi, U.S. and Nzelibe, C.H. (2012). Purification and characterization of a cysteine protease from the bulb of common onion Allium cepa L. International Journal of Modern Biochemistry 1(1); 1-17.
-  Ogbonna, A., Obi S., Okolo, B. and Odibo, F. (2003). Purification and some properties of a protease from sorghum malt variety KSVB-11. Journal, Institute of Brewing 109; 179-186.
-  Palma, J.M., Sandalio, L.M., Corpas, F.J., Romero-Puertas, M.C., McCarthy, I. and Del Rio, L.A. (2002). Plant proteases, protein degradation, and oxidative stress: role of peroxisomes. Plant Physiology and Biochemistry 40; 521-530.
-  Pandhare, J., Zog, K. and Deshpande, V.V. (2002). Differential stabilities of alkaline protease inhibitors from actinomycetes: effect of various additives on thermostability. Bioresource Technology 84; 165-169.
-  Patel, A.K., Singh, V.K. and Jagannadham, M.V. (2007). Carnein: A serine protease from noxious plant weed Ipomoea carnea (Morning glory). Journal of Agricultural Food Chemistry 55; 5809-5818.
-  Perpetuo, E.A., Juliano, L. and Lebrun, I. (2003). Biochemical and pharmacological aspects of two bradykinin- potentiating peptidases obtained from tryptic hydrolysis of casein. Journal of Protein Chemistry 22; 601-606.
-  Qi, H., Dong, P.X., Cong, N.L., Gao, Y., Liu, L., Mikiro, T. and Zhu, B.W. (2007). Purification and characterization of a cysteine-like protease from the body wall of the sea cucumber Stichopus japonicas. Fish Physiology and Biochemistry 33; 181-188.
-  Raimi, O.G., Elemo, B.O., Fatai, A.A., Bankole, H.A., Kazeem, M.I. and Banjoko, A.O. (2011). Isolation and partial characterization of a protease enzyme from Thaumatococcus daniellii waste. African Journal of Biotechnology 10 (16); 3186-3190.
-  Rajeswari, J., Vadde, R., Sridhar, R. and Ramakrishna, R. (2009). Purification and characterization of cysteine protease from germinating cotyledons of horse gram. BMC Biochemistry 10: 1-10.
-  Rao, M.B., Tanksale, A.M., Ghalge, M.S. and Deshpande, W. (1998). Molecular and Biotechnological aspects of microbial proteases. Microbiology and Molecular Biology Review 62; 597-635.
-  Sareen, R. and Mishra, P. (2008). Purification and characterization of organic solvent stable protease from Bacillus licheniformis RSP-09-37. Applied Microbiology and Biotechnology 79; 399-405.
-  Shankar, S., Rao, M. and Laxman, R.S. (2011). Purification and characterization of an alkaline protease by a new strain of Beauveria sp. Process Biochemistry 46; 579-585.
-  Silva, S.V. and Malcata, F.X. (2005). Partial identification of water soluble peptides released at early stages of proteolysis in sterilized ovine cheese-like systems: influence of type of coagulant and starter. Journal of Dairy Science 88; 1947-1954.
-  Sumantha, A., Sandhya, C., Szakacs, G., Soccol, C.R. and Pandey, A. (2005). Production and partial purification of a neutral metalloprotease by fungal mixed substrate fermentation. Food Technology and Biotechnology 43 (4); 313-319.
-  Thangam, B.E. and Rajkumar, S.G. (2002). Purification and characterization of alkaline protease from Alcaligenes faecalis. Biotechnology and Applied Biochemistry 35; 149-154.
-  Vishwanatha, K.S., Appu, R.A.G. and Sigh, S.A. (2009). Characterization of acid protease expressed from Aspergillus oryzae MTCC 5341. Food. Chemistry 114; 402-407.
-  Walsh, G. (2002). Proteins, Biochemistry and Biotechnology. West Susex, England: John Wiley and Sons Ltd pp. 420.
-  Yadav, S.C., Pande, M. and Jagannadham, M. (2006). Highly stable glycosylated serine protease from the medicinal plant Euphorbia milii. Phytochemistry 67; 1414-1426.
-  Zhang, N. and Jones, B.L. (1996). Purification and partial characterization of a 31 kDa cysteine endopeptidase from germinated barley. Planta 199: 565-572.
-  Zhang, X., Liu, Q., Zhang, G., Wang and Hand, N. T, (2010). Purification and molecular cloning of a serine protease from the mushroom Hypsizigus marmoreus. Process Biochemistry 45; 724-730.
Publication order reference