Response surface methodology optimization of milk-clotting protease produced by Pleurotus sajor-caju strain CTM 10057 and its technico-economical evaluation

M. Omrane Benmrad, W. Gargouri, D. Mahfoudh, M. Kriaa, B. Jaouadi, N. Kechaou


Abstract:The article purpose is to scale-up the bioreactor design for the serine protease from Pleurotus sajor-caju strain CTM 10057 (called SPPS) production to be profitably used in cheese-making production. The SPPS enzyme has been optimized using central composite design (CCD) statistical design analysis. The optimal conditions are: a carbon source of 15 g/L, a nitrogen source of 2 g/L, agitation of 160 rpm at pH = 5.6. This production is scaled-up using submerged fermentation. Under the optimized conditions, protease yield 75,000 U/mL was 6.5 folds higher than those obtained by the use of the initial conditions (11,600 U/mL). The separation was carried out throughout centrifugation followed by a concentration with evaporation, after which this enzyme was tested in the enzymatic milk coagulation process. According to its milk-clotting ability, it could be used in the cheese industry, as well as other food industries. Interestingly, the economic analysis of this planned bioprocess is leased at a positive gain. The estimated SPPS price will be competitive in comparison with commercial enzymes (Mucorpepsine and Presura). So, SPPS enzyme can be used in cheese making and its process is profitable.

Full Text:



Turton, R.; Bailie, R.C.; Whiting, W.B.; Shaeiwitz, J.A. Analysis, Synthesis, and Design of Chemical Processes. Fourth Edition, Pearson Education, Inc., The Prentice Hall International Series in the Physical and Chemical Engineering Sciences (1998) 1-108.

Ringborg, R.H.; Woodley, J. The application of reaction engineering to biocatalysis. Reaction Chemistry & Engineering 1 (2016) 10-22.

Charpentier, J.C. Among the trends for a modern chemical engineering, the third paradigm: The time and length multiscale approach as an efficient tool for process intensification and product design and engineering. Chemical Engineering Research and Design 88 (2010) 248-254.

Charpentier, J.C.; Brain natriuretic peptide: A marker of myocardial dysfunction and prognosis during severe sepsis.Critical Care Medicine 32 (2004) 660-665.

Chakraborty, A. Gopalakrishnan, S.; Reddy, J. A new beam finite element for the analysis of functionally graded materials. International Journal of Mechanical Sciences 45 (2003) 519-539.

Silva, S.V.; Malcata, F.X. Studies pertaining to coagulant and proteolytic activities of plant proteases from Cynara cardunculus. Food Chemistry 89 (2005) 19-26.

Arnaud, A.J.; C.d.S.E.N.; Juridiques, Dictionnaire Encyclopédique de Théorie et de Sociologie du Droit. Anthologie du droit 1re édition (1993).

Dang, T.P. Catalyse d'hydrogénation en phase homogène des aldéhydes α-β insatures. Application a la synthèse asymétrique du citronellal.Journal of Molecular Catalysis, 16 (1982) 51-59.

Aunstrup, K. Proteinases in Economic Microbiology: Microbial Enzymes and Bioconversions 5 (1980) 50-114.

Arima, K.; Yu, J.; Iwasaki, S. Milk-clotting enzyme from Mucor pusillus var. Lindt, in Methods in Enzymology, 19 (1970) 446-449.

Granday, P. Aptitude technologique de différents coagulants du lait. Influence de la composition de leur" support". Le Lait,67(1987) 187-218.

Marcone, M.; Kakuda, Y.; A comparative study of the functional properties of amaranth and soybean globulin isolates. Molecular Nutrition & Food Research 43 (1999) 368-373.

AFNOR, E.; Guide général pour la sélection, l'entraînement et le contrôle des sujets. (1994) 11.

Rose, A.H.; Microbial Enzymes and Bioconversions. (1980).

Kalk, J.P.; Langlykke, A. Cost estimation for biotechnology projects. (1986) 363-385.

Brack, M. Funny hills: The shell-correction approach to nuclear shell effects and its applications to the fission process.Reviews of Modern Physics 44 (1972) 320.

Ramasubramanian, L.; Webb, R.; D’Arcy B.; Deeth, H.C.; Characteristics of a calcium–milk coagulum. Journal of Food Engineering 114 (2013) 147-152.

Aswal, P.; Shukla, A.; Priyadarshi S.; Yoghurt: Preparation, characteristics and recent advancements. Cibtech Journal of Bio-Protocols 1 (2012) 32-44.

Vyhmeister, S. Influence of using different proportions of cow and goat milk on the chemical, textural and sensory properties of Chanco–style cheese with equal composition. LWT - Food Science and Technology 112 (2019) 108-226.

Rodrigo, A. Ibáñez, R.; Waldron, D.; McSweeney, P.L.H.; Effect of fat content and temperature on the translucency of Cheddar cheese. International Dairy Journal 54 (2016) 33-42.

Aboushanab, S.; Vyrova D.; Selezneva I.; Characterization of low- and non-fat yogurt manufactured with addition of beta-glucanas a dietary supplement, 1 (2018) 2015. AIP Conference Proceedings 2015, 020003 (2018);


  • There are currently no refbacks.

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License. Copyright UMMB © 2020 University M'hamed Bougara - Boumerdes. Independance AV., Boumerdes, 35000 Algeria, Tel/Fax: +213 24 91 14 98/ +213 2491-29-51