Production and purification of milk-clotting protease produced by local fungal Mucor sp. in solid cultures

S. Bensmail; F. Lahouari; F.Z. Kouadri; S. Talantikit; F. Fazouane

Production and purification of milk-clotting protease produced by local fungal Mucor sp. in solid cultures

In: Industrial, medical and environmental applications of microorganisms

Authors:

S. Bensmail

S. Bensmail ¹Research Laboratory of Food Technology, Faculty of Engineering, Department of Food Technology, M’Hamed Bougara University, Independence Street, 35000 Boumerdes, Algeria; souhilabensmail@yahoo.fr; ffazouane@yahoo.fr
²Laboratory of Biochemistry, Department of Biology, Faculty of Sciences, M’Hamed Bougara University, Independence Street, 35000 Boumerdes, Algeria

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F. Lahouari

F. Lahouari ²Laboratory of Biochemistry, Department of Biology, Faculty of Sciences, M’Hamed Bougara University, Independence Street, 35000 Boumerdes, Algeria

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F.Z. Kouadri

F.Z. Kouadri ²Laboratory of Biochemistry, Department of Biology, Faculty of Sciences, M’Hamed Bougara University, Independence Street, 35000 Boumerdes, Algeria

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S. Talantikit

S. Talantikit ¹Research Laboratory of Food Technology, Faculty of Engineering, Department of Food Technology, M’Hamed Bougara University, Independence Street, 35000 Boumerdes, Algeria; souhilabensmail@yahoo.fr; ffazouane@yahoo.fr

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, and

F. Fazouane

F. Fazouane ¹Research Laboratory of Food Technology, Faculty of Engineering, Department of Food Technology, M’Hamed Bougara University, Independence Street, 35000 Boumerdes, Algeria; souhilabensmail@yahoo.fr; ffazouane@yahoo.fr
²Laboratory of Biochemistry, Department of Biology, Faculty of Sciences, M’Hamed Bougara University, Independence Street, 35000 Boumerdes, Algeria

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Type:: Chapter

Pages:: 362–368

DOI:: https://doi.org/10.3920/9789086867950_064

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The worldwide increase in cheese production and the reduced supply of calf rennet, have led to an increase in the demand for alternative sources of milk coagulants. Various proteases have been suggested to be used as calf rennet substitutes, where more attention has been focused on the production of milk-clotting enzymes from microbial sources. The aim of our study is the production and purification of a milkclotting protease produced by local fungal strain Mucor sp. isolated from the soil, grown on solid medium containing wheat bran. The maximum enzyme productivity (4,800 US/g wheat bran; proteins 2.3 mg/g wheat bran) was obtained under the optimum conditions of temperature (25 °C), incubation time (96 h), moisture content of solid substrate (43%) adjusted suitably with mineral solution (Czapek-Dox) of pH 3. A fold purification of 4.827 was obtained after precipitation with ammonium sulfate at an optimum rate of 60% compared to those obtained at 40% (0.4) and 80% (2.527). Size exclusion chromatography on Sephadex G-100 of the dialyzed extract provides an increase in the fold purification to a value of 15.42. SDS-PAGE revealed that the purified enzyme is monomeric with a molecular weight of 36 kDa.

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Industrial, medical and environmental applications of microorganisms

Current status and trends

E-Book ISBN:: 9789086867950

Publisher:: Wageningen Academic

Print Publication Date:: 01 Nov 2014

Subjects
- Life Sciences

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Production and purification of milk-clotting protease produced by local fungal Mucor sp. in solid cultures

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Industrial, medical and environmental applications of microorganisms

Current status and trends

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