Author(s): Saurabh Singh, Shivangi Ahuja, Lakshmi Thangjam, Neha Kumari, Daljeet Singh Dhanjal, Chirag Chopra, Reena Singh


DOI: 10.5958/0974-360X.2021.00038.X   

Address: Saurabh Singh1, Shivangi Ahuja1, Lakshmi Thangjam1, Neha Kumari1, Daljeet Singh Dhanjal1, Chirag Chopra1, Reena Singh1*
1School of Bioengineering and Biosciences, Lovely Professional University, Punjab-144411, India.
*Corresponding Author

Published In:   Volume - 14,      Issue - 1,     Year - 2021

Proteases, the enzymes that digest proteins, are present in all organism and play an essential role in proliferation and differentiation. They are crucial for different applications in industries like detergent, food, laundry and pharmaceutical. Proteases from bacteria and fungi have emerged as a salvation to the industrial sectors. In the presented work, protease producing Aspergillus terreus was grown from the field soil of Birring village, Jalandhar. Further, Aspergillus terreus was screened for growth on different substrates like Okara, Broken Wheat, Chick Pea and Black Gram. After, substrate selection, the solid-state fermentation was found to be an efficient method for enzyme production. Upon characterisation, the crude enzyme was found to be extracellular. The optimum enzyme activity was at pH of 10.0 and a temperature of 55oC. Also, Mn2+enhanced the enzyme activity. Thus, the low-cost substrate and effective protease activity, can make this enzyme as ideal candidate for various industrial applications.

Cite this article:
Saurabh Singh, Shivangi Ahuja, Lakshmi Thangjam, Neha Kumari, Daljeet Singh Dhanjal, Chirag Chopra, Reena Singh. Comparative Evaluation of Protease Production from Okara, Broken wheat, Chickpea and black gram by the soil isolate of Aspergillus terreus. Research J. Pharm. and Tech. 2021; 14(1):219-224. doi: 10.5958/0974-360X.2021.00038.X

Saurabh Singh, Shivangi Ahuja, Lakshmi Thangjam, Neha Kumari, Daljeet Singh Dhanjal, Chirag Chopra, Reena Singh. Comparative Evaluation of Protease Production from Okara, Broken wheat, Chickpea and black gram by the soil isolate of Aspergillus terreus. Research J. Pharm. and Tech. 2021; 14(1):219-224. doi: 10.5958/0974-360X.2021.00038.X   Available on:

1.    Panesar PS, Kaur R, Singla G and Sangwan RS. Bio-processing of agro-industrial wastes for production of food-grade enzymes: progress and prospects. Applied Food Biotechnology. 2016; 3(4): 208-27.
2.    Chand T, Pandey FK, Dhingra S, Sharma MK. Production of Industrially Significant Enzymes from Bio-Wastes Using Aspergillus niger by Solid State Fermentation. Research Journal of Pharmaceutical Dosage Forms and Technology. 2014;6(1):33-6.
3.    Utharalakshmi N, Kumar AG, Narendra kumar G. Optimization of Cellulase Producing Aspergillus flavus SB4 by Solid State Fermentation using Response Surface Methodology (RSM)-CCD. Research Journal of Pharmacy and Technology. 2015;8(4):349-54.
4.    Jagathy K, Kalpana K, Rajesh kumar S. Production, optimization, Characterization and Immobilization of Glucose oxidase from Aspergillus species. Research Journal of Pharmacy and Technology. 2017;10(6):1924-8.
5.    Mótyán J, Tóth F and Tőzsér J. Research applications of proteolytic enzymes in molecular biology. Biomolecules. 2013; 3(4): 923-42.
6.    Robinson PK. Enzymes: principles and biotechnological applications. Essays in Biochemistry. 2015; 59: 1-41.
7.    Ravindran R and Jaiswal A. Microbial enzyme production using lignocellulosic food industry wastes as feedstock: a review. Bioengineering. 2016; 3(4): 30.
8.    Chanda I, Chanda SR, Dutta SK. Anti-inflammatory Activity of a Protease Extracted from the Fruit Stem Latex of the Plant Artocarpus heterophyllus Lam. Research Journal of Pharmacology and Pharmacodynamics. 2009;1(2):70-2.
9.    Shahina Z, Hossain MT and Hakim MA. Variation of Protease Production by the Bacteria (Bacillus fastidiosus) and the Fungus (Aspergillus funiculosus). Journal of Microbiology Research. 2013: 135-42.
10.    Nigam P. Microbial enzymes with special characteristics for biotechnological applications. Biomolecules. 2013; 3(3): 597-611.
11.    Benluvankar V, Jebapriya GR and Gnanadoss JJ. Protease production by Penicillium sp. LCJ228 under solid state fermentation using groundnut oilcake as substrate. Life. 2015; 50: 12.
12.    Muthukumaran PM, Mathumitha M. Decolorization and degradation of reactive dyes by Aspergillus niger. Research Journal of Engineering and Technology. 2013;4(4):235-8.
13.    Singh S, Farooq D, Thakur R. Screening of Natural Rubber Degradation by Fungi; Aspergillus and Phlebia Sp. and Bacteria Pseudomonas and Streptomyces sp. Research Journal of Pharmacy and Technology. 2017;10(11):3939-44.
14.    Chatterjee A, Abraham J. Biosorption of Copper using Oryza sativa and Aspergillus oryzae. Research Journal of Pharmacy and Technology. 2016;9(6):664-70.
15.    Wu TY, Mohammad AW, Jahim JM and Anuar N. Investigations on protease production by a wild-type Aspergillus terreus strain using diluted retentate of pre-filtered palm oil mill effluent (POME) as substrate. Enzyme and Microbial Technology. 2006; 39(6):1223-9.
16.    Gao J, Weng H, Zhu D, Yuan M, Guan F and Xi Y. Production and characterization of cellulolytic enzymes from the thermoacidophilic fungal Aspergillus terreus M11 under solid-state cultivation of corn stover. Bioresource Technology. 2008; 99(16): 7623-9.
17.    Sethi BK, Rout JR, Das R, Nanda PK and Sahoo SL. Lipase production by Aspergillus terreus using mustard seed oil cake as a carbon source. Annals of Microbiology. 2013; 63(1): 241-52.
18.    Souza PM, Bittencourt ML, Caprara CC, Freitas MD, Almeida RP, Silveira D, Fonseca YM, Ferreira Filho EX, Pessoa Junior A and Magalhães PO. A biotechnology perspective of fungal proteases. Brazilian Journal of Microbiology. 2015; 46(2): 337-46.
19.    Jain P, Pundir RK. Antagonistic activity of soil fungal metabolite against Streptococcus mutans strains. Research Journal of Pharmacy and Technology. 2010;3(2):417-9.
20.    Cappuccino JG and Sherman N. Microbiology: a laboratory manual. San Francisco: Pearson/Benjamin Cummings; 2005.
21.    Balajee SA, Houbraken J, Verweij PE, Hong SB, Yaghuchi T, Varga J and Samson RA. Aspergillus species identification in the clinical setting. Studies in Mycology. 2007; 59: 39-46.
22.    Kamath P, Subrahmanyam VM, Rao JV and Raj PV. Optimization of cultural conditions for protease production by a fungal species. Indian Journal of Pharmaceutical Sciences. 2010; 72(2): 161.
23.    Almalki MA. Solid state fermentation of agro-residues for the production of amylase from Bacillus subtilis for industrial applications. International Journal of Current Microbiology and Applied Sciences. 2018; 7(3): 1341-8.
24.    Radha S, Sridevi A, Himakiran Babu R, Nithya VJ, Prasad NB and Narasimha G. Medium optimization for Acid protease production from Aspergillus sp under solid state fermentation and mathematical modelling of protease activity. Journal of Microbiology and Biotechnology Research. 2012 ;2: 6-16.
25.    Paranthaman R, Alagusundaram K and Indhumathi J. Production of protease from rice mill wastes by Aspergillus niger in solid state fermentation. World Journal of Agricultural Sciences. 2009; 5(3): 308-12.
26.    Vijayaraghavan P and Vincent SG. A simple method for the detection of protease activity on agar plates using bromocresol green dye. Journal of Biochemical Technology. 2013; 4(3): 628-30.
27.    Oliveira AN, Oliveira LA and Andrade JS. Production and some properties of crude alkaline proteases of indigenous Central Amazonian Rhizobia strains. Brazilian Archives of Biology and Technology. 2010; 53(5):1185-95.
28.    Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry. 1976; 72(1-2): 248-54.
29.    Classics Lowry O, Rosebrough N, Farr A and Randall R. Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry. 1951; 193: 265-75.
30.    Fan Y, Tian L, Xue Y, Li Z, Hou H and Xue C. Characterization of protease and effects of temperature and salinity on the biochemical changes during fermentation of Antarctic krill. Journal of the Science of Food and Agriculture. 2017; 97(11): 3546-51.
31.    Vidhya CV. Production and Optimization of Extra-cellular protease from Ganoderma sp. Research Journal of Pharmacy and Technology. 2019;12(4):1832-8.
32.    Gaddeyya G, Niharika PS, Bharathi P and Kumar PR. Isolation and identification of soil mycoflora in different crop fields at Salur Mandal. Advances in Applied Science Research. 2012; 3(4): 2020-6.
33.    Pilli R and Siddalingeshwara KG. Rapid confirmation and molecular identification of alkaline protease producing Aspergillus awamori through submerged fermentation. International Journal of Current Microbiology and Applied Sciences. 2016; 5: 1114-24.
34.    Rao GG, Patel PM, Rao BV and Pancha RR. Alkaline protease production by Aspergillus terreus BAB-346 using poultry litter waste. International Journal of Current Microbiology and Applied Sciences. 2016; 5(10): 174-84.
35.    Sethi BK, Jana A, Nanda PK, Das Mohapatra PK and Sahoo SL. Thermostable acidic protease production in Aspergillus terreus NCFT 4269.10 using chickling vetch peels. Journal of Taibah University for Science. 2016; 10(4): 571-83.
36.    Rai MP and Nigam S. Cost Effective Method of Protease Production in Solid State Fermentation Using Combined Substrate Corn Cob and Lentil Husk. Research and Reviews: A Journal of Life Sciences, 2018; 3(1): 1-7.
37.    El-Safey EM and Abdul-Raouf UM. Production, purification and characterization of protease enzyme from Bacillus subtilis. In: International Conferences for Development and the Environment in the Arab World 2004 (p. 14). Assiut University.
38.    Archna S, Priyank V, Nath YA and Kumar SA. Bioprospecting for extracellular hydrolytic enzymes from culturable thermotolerant bacteria isolated from Manikaran thermal springs. Research Journal of Biotechnology. 2015; 10(4): 33-42.
39.    Jugran J, Joshi GK, Bhatt JP and Shanker A. Production and partial characterization of extracellular protease from Bacillus sp. GJP2 isolated from a hot spring. Proceedings of the National Academy of Sciences, India Section B: Biological Sciences. 2016; 86(1): 171-8.
40.    Sachan S, Iqbal MS and Singh A. Extracellular lipase from Pseudomonas aeruginosa JCM5962 (T): Isolation, identification, and characterization. International Microbiology. 2018; 21(4): 197-205.
41.    Gomaa EZ. Optimization and characterization of alkaline protease and carboxymethyl-cellulase produced by Bacillus pumillus grown on Ficus nitida wastes. Brazilian Journal of Microbiology. 2013; 44(2): 529-37.
42.    Agrawal D, Patidar P, Banerjee T and Patil S. Production of alkaline protease by Penicillium sp. under SSF conditions and its application to soy protein hydrolysis. Process biochemistry. 2004; 39(8): 977-81.
43.    Agrawal D, Patidar P, Banerjee T and Patil S. Alkaline protease production by a soil isolate of Beauveria felina under SSF condition: parameter optimization and application to soy protein hydrolysis. Process Biochemistry. 2005; 40(3-4): 1131-6.
44.    Sumantha A, Sandhya C, Szakacs G, Soccol CR and Pandey A. Production and partial purification of a neutral metalloprotease by fungal mixed substrate fermentation. Food Technology and Biotechnology. 2005; 43(4): 313-9.
45.    Sharma KM, Kumar R, Panwar S and Kumar A. Microbial alkaline proteases: Optimization of production parameters and their properties. Journal of Genetic Engineering and Biotechnology. 2017; 15(1): 115-26.
46.    De Castro RJ and Sato HH. Protease from Aspergillus oryzae: biochemical characterization and application as a potential biocatalyst for production of protein hydrolysates with antioxidant activities. Journal of Food Processing. 2014; 2014.
47.    Gaur S, Agrahari S and Wadhwa N. Purification of protease from Pseudomonas thermaerum GW1 isolated from poultry waste site. The Open Microbiology Journal. 2010; 4: 67-74.
48.    Sitoke A, Chopra RS, Kumar GP, Chopra C. Identification and Characterization of an Alkalophilic Protease from Bacillus Mycoides strain isolated from Industrial Soil of Phagwara, India. Research Journal of Pharmacy and Technology. 2017;10(10):3435-8.

Recomonded Articles:

Research Journal of Pharmacy and Technology (RJPT) is an international, peer-reviewed, multidisciplinary journal.... Read more >>>

RNI: CHHENG00387/33/1/2008-TC                     
DOI: 10.5958/0974-360X 

56th percentile
Powered by  Scopus

SCImago Journal & Country Rank

Recent Articles


Not Available