STATISTICAL AND KINETIC STUDIES OF ACID PROTEASE BY ASPERGILLUS SPP. ISOLATED FROM SOIL CONTAMINATED WITH ABATTOIR WASTE


Radha S., Sridevi A., Prasad Nbl, Narasimha G.

Abstract


Objective: Aim of the present investigation was to optimize the acid protease production from Aspergillus spp. through statistical method in solid state fermentation and to study the inhibitory enzyme kinetics.

Methods: To fulfill above mentioned aim, seven solid substrates were screened though using PBD (Plackett-Burman Design) and concentrations of three significant were determined by using one of the Response surface methodologies (RSM), Box-Behnken design (BBD). Inhibitory enzymatic effects were carried by using previously developed models.

Results: From PBD, wheat bran, soybean meal, and dried potato peel (DPP) were screened as major influencing nutritional factors for enzyme production. Better optimal values were determined by BBD as wheat bran: 8.841 g, soybean meal: 4.557 g, and DPP: 0.661 g with predicted protease activity as 817.83 U/g (±44.047 U/g). Linear, interactive, and quadratic effects of aforesaid substrates on enzyme activity were formulated by quadratic model through multiple regression model (R2Adj:Adjusted R square = 94.78%; R2Pre:Predicted R square = 98.13%). Partial substrate inhibition to crude acid protease activity was notified with casein concentration higher than 0.4 mmol and inhibitory constant, KN, was computed with previous developed mathematical models. Ratio of reaction rate constants, k4/k2, was found to be 0.233 that had confirmed partial casein inhibition to enzyme velocity. Improved activity and kinetics of caseinolysis make amicable for industrial applications.

Conclusion: Quick optimization was performed with statistical methodology over conventional approach. Inhibitory enzyme kinetic studies were important for industrial applications of acid protease.


Keywords


Acid protease, Optimization, Statistical methodology, Casein inhibition, Reaction velocity

| PDF | HTML |

References


Azzam A, Ramzi AAA, Avnish P, Nor Zalina O, Roslinda AM, El Enshasy HA. Biotechnological aspects and pharmaceutical applications of bacterial proteases. Pharm Lett 2017;9:9-20.

Rao MB, Tanksale AM, Ghatge MS, Deshpande VV. Molecular and biotechnological aspects of microbial proteases. Microbiol Mol Biol Rev 1998;62:597-635.

Souza PM, Bittencourt MLA, Caprara CC, Freitas M. Biotechnology perspective of fungal proteases–a review. Braz J Microbiol 2015;46:337-46.

Venkat KS, Ashok Rao, Joshua RN. Screening, media optimization and partial purification of protease by Trichosporonjaponicum VITVK1. Int J Pharm Pharm Sci 2015; 7:187-91.

Kalaskar VV, Narayanan K, Subrahmanyam VM, Rao VJ. Partial characterisation and therapeutic application of protease from a fungal species. Indian Drugs 2012;49:42-6.

Viswanatha KS, Appu Rao AG, Singh SA. Acid protease production using solid-state fermentation by Aspergillus oryzae MTCC 5341: optimization of process parameters. J Indian Microbial Biotechnol 2009;37:129-38.

Siala R, Frikha F, Mhamdi S, Nasri M, Sellami AK. Optimization of acid protease production by Aspergillus niger I1 on shrimp peptone using statistical experimental design. Sci World J 2012;11. DOI:10.1100/2012/564932.

Djamel C, Ali T, Nelly C. Acid protease production by isolated species of Penicillium. Eur J Sci Res 2009;25:469-77.

Soares VF, Castilho LR, Bon EP, Freire DM. High-yield Bacillus subtilis for protease production by solid-state fermentation. Appl Biochem Biotechnol 2005;121-124:311-9.

Aravindhan R, Anbumathi P, Viruthagiri T. Evaluation of medium components by Plackett-Burman statistical design for lipase production by Candida rugosa and kinetic modeling. Chinese J Biotechnol 2008;24:436-44.

Vasanthabharathi V, Shellarani S, Lakshminarayanan R, Jayalakshmi S. Statistical optimization and production of protease from estuarine Citrobacterdiversus. Int Res J Biol Sci 2016;5:18-25.

Xiong ZQ, Tu XR, Tu GQ. Optimization of medium composition for actinomycin X2 production by Streptomyces spp. JAU4234 using response surface methodology. J Indian Microbiol Biotechnol 2008;35:729–34.

Wang Q, Yanhua H, Zhong X, Jinlai M, Guangyou L. Optimization of cold-active protease production by the psychrophilic bacterium Colwellia sp. with response surface methodology. Bioresour Technol 2008;99:1926-31.

Hong G, Mei L, Jintao L, Huanqin D, Xianlong Z, Xiangyang L, et al. Medium optimization for the production of avermectin B1 a by Streptomyces avermitilis 14-12A using response surface methodology. Bioresour Technol 2009;100:4012-6.

Asis KM, Sudir KR. A statistical approach for the enhanced production of alkaline protease showing the fibrinilytic activity from a newly isolated gram-negative Bacillus sp. strain AS-S20-I. New Biotechnol 2011;28:182-9.

Reed MC, Anna L, Frederik NH. The biological significance of substrate inhibition: a mechanism with diverse functions. Bioessays 2010;32:422–9.

Nejad ZG, Soheila Y, Nazanin M, Bahareh S. Some investigations on protease enzyme production kinetics using Bacillus licheniformis BBRC 100053 and effects of inhibitors on protease activity. Int J Chem Engg 2014. http://dx.doi.org/ 10.1155/2014/394860

Yoshino M, Murakam K. Analysis of the substrate inhibition of complete and partial types. Springerplus 2015;4:292.

Radha S, Sridevi A, Himakiran BR, Nithya VJ, Prasad NBL, Narasimha G. Isolation and screening of proteolytic fungal cultures from soil contaminated with abattoir waste. Biochem BCAIJ 2012;6:226-30.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with folin phenol reagent. J Biol Chem 1951; 193:265-75.

Shuler ML, Kargi F. Enzymes. A text book of Bioprocess Engineering–Basic Concepts. 2nd Edition. Pearson Education: India; 2003. p. 60-73.

Okpokwasili GC, Nweke CO. Microbial growth and substrate utilization kinetics. Afr J Biotechnol 2005;5:305-7.

Lu-E Shi, Guo-Qing Y, Xiao-Ying Z, Zhen-Xing T, Jian-Shu C, Wen-Yue X, et al. Medium optimization for 5’-Phospshodiesterase production from Penicillium citrinumusing response surface methodology. Food Technol Biotechnol 2007;45:126-33.

Plackett RL, Burman JP. The design of optimum multifactorial experiments. Biometrica 1946;33:305-25.

Radha S, Sridevi A, Himakiran Babu R, Nithya VJ, Prasad NBL, Narasimha G. Medium optimization for acid protease production from Aspergillus sps. under solid state fermentation and mathematical modeling of protease activity. J Microbiol Biotechnol Res 2012;2:6-16.

Chauhan B, Gupta R. Application of statistical experimental design for optimization of alkaline protease production from Bacillus sp. RGR-14. Process Biochem 2004;39:2115–22.

Rajshree S, Rajni S. Statistical optimization of conditions for protease production from Bacillus sp. Acta Biologica Szegediensis 2014;54:35-141.

Govarthanan M, Lee J, Cho M, Seralathan K, Byung-Taek, O. Statistical optimization of alkaline protease production from brackish environment Bacillus sp. SKK 11 by SSF using horse gram husk. Preparative Biochem Biotechnol 2014;44:119–31.

Xiao Y, Wu D, Zhao S, Lin W, Gao X. Statistical optimization of alkaline protease production from Penicillium citrinumYL-1 under solid-state fermentation. Preparative Biochem Biotechnol 2015;45:447–62.

Kshetri P, Ningombam O, Ningombam DS. Optimization of alkaline protease production by alkaliphilic Bacillus sp. KW2 in low cost medium using statistical approaches. Appl Microbiol 2016;2:2.

Rathakrishnan P, Nagarajan P, Rajeshkannan R. Optimization of protease production by Bacillus licheniformisin sugarcane bagasse using statistical experimental design. Resin Biotechnol 2012;3:1-10.

Qureshi AS, Dahot UM. Production of proteases by Staphylococcus epidermidis EFRL using cost effective substrate (molasses) as a carbon source. Pakistan J Biotechnol 2009;6:55-60.

Kandasamy S, Muthusamy G, Balakrishnan S, Duraisamy S, Thangasamy S. Optimization of protease production from surface-modified coffee pulp waste and corncobs using Bacillus sp. by SSF. 3 Biotech 2016;6:167.

Niyonzima FN, More SS. Purification and characterization of detergent-compatible protease from Aspergillus terreus gr. 3 Biotech 2015;5:61–70.




About this article

Title

STATISTICAL AND KINETIC STUDIES OF ACID PROTEASE BY ASPERGILLUS SPP. ISOLATED FROM SOIL CONTAMINATED WITH ABATTOIR WASTE

Keywords

Acid protease, Optimization, Statistical methodology, Casein inhibition, Reaction velocity

DOI

10.22159/ijpps.2018v10i2.23165

Date

01-02-2018

Additional Links

Manuscript Submission

Journal

International Journal of Pharmacy and Pharmaceutical Sciences
Vol 10, Issue 2, 2018 Page: 72-79

Online ISSN

0975-1491

Authors & Affiliations

Radha S.
Department of Biotechnology, Sree Vidyanikethan Engg. College, Tirupati
India

Sridevi A.
Department of Microbiology, Mahila University, Tirupati, Andhra Pradesh
India

Prasad Nbl
JNTUA, OTPRI, Ananthapuramu, Andhra Pradesh, India
India

Narasimha G.
Applied Microbiology Lab, Department of Virology, Sri Venkateswara University, Tirupati, Andhra Pradesh
India


Article Tools



Refbacks

  • There are currently no refbacks.