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Partial Purification of Deamidase from Germinating Wheat (triticum Aestivum L.) Seeds and Its Application for the Improvement of Functional Properties of Wheat Gluten

M M Rahman
Department of Applied Nutrition & Food Technology, Islamic University of Kustia

N K Sana
Department of Biochemistry & Molecular Biology, Islamic University of Kustia

M M H Khan
Department of Biochemistry & Molecular Biology, Islamic University of Kustia

B C Sarkar
Department of Biochemistry & Molecular Biology, Islamic University of Kustia

E M Huque
Department of Chemistry, University of Rajshahi, Rajshahi-6205, Bangladesh

Shaha Ranajit Kumar
Department of Biochemistry & Molecular Biology, Islamic University of Kustia

Abstract/ Executive Summary:

Enzyme with deamidase activity was found in the germinating wheat seeds. The activity appeared in the seeds after 24 hours of germination and reached its maximum value after 48 hours and then declined rapidly. Protein deamidase from germinating wheat seeds was further purified by salting out with phosphate salts and by subsequent chromatography on gel filtration and DEAE-cellulose column chromatography. Polyacrylamide slab gel electrophoresis showed the purified enzyme to be homogeneous. The molecular weight was determined to be 59 kDa by gel filtration and 60 kDa, by SDS- Polyacrylamide slab gel electrophoresis (SDS-PAGE). The optimum pH and temperature of the purified enzyme were found to be 6.9 and 32°C, respectively. The functional properties of wheat gluten were investigated after treating with it purified deamidase enzyme from germinating wheat seeds at pH 6.9 at 32°C. The functional properties such as solubility, emulsification and foam formation of deamidase treated gluten were improved greatly as compared to that of native gluten. The solubility of the deamidase treated gluten was remarkably high in the pH range of 6 to 8, in which the native gluten was insoluble. It was apparent that the improvement of functional properties of wheat gluten was mainly due to the deamidation induced by wheat deamidase at pH 6.9 and 32°C.

Keywords: Wheat seeds, Deamidation, Wheat gluten, DEAE-cellulose

Location: Bangladesh Agriculture Research Institute, Substation of Ishwardi, Pabna.

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Wheat

Objectives:

To the purification of deamidating enzyme (deamidase) from germinating wheat seeds and.improvernent of functional properties (solubility, emulsifying and foaming) of wheat gluten . by treating with purified deamidase from germinating wheat seeds.

Methodology:

Wheat (Triticum aestivum,L.) seeds were collected from Bangladesh Agriculture research Institute (BARI), substation of Ishwardi, pabna. The seeds were soaked in distilled water for 6 hrs, germinated in the dark at 25°C for 24h, 48h, 72h and 96h including soaking time. The germinated seeds at different hours stored separately in the deep freeze (-10°C) for further experimental purpose. Twenty grams of germinated wheat seeds (germinated at 48 h) were taken in a pre-cooled mortar and pasted with a pestle and homogenized with cold de-ionized distilled water in a ratio of 3:5 (weight of dry seeds/water volume) and then centrifuged at 6x10³rpm (at 2°C) for 10 minutes by a refrigerated centrifuge machine. After centrifugation, the clear supernatant solution (water extract) was taken as a crude extract of the enzyme. The enzymatically active protein fractions after gel filtration were collected Working substrate globulins was prepared from wheat seeds by the method of Thanh and Shibasaki. An insoluble wheat protein complex, gluten, was prepared from flour dough washed until the washings were free from soluble protein. The gluten ball thus obtained was dialyzed against distilled water and then freeze- dried in a freeze-dryer.

Source of Information: Journal of Bio-Science. Vol. 15, pp: 89-98, 2007

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

It was observed that by treating with deamidase at pH 6.9 the deamidated gluten could be obtained. The deamidated gluten greatly improved some functional properties such as solubility, emulsifying and foaming ability. It was suggested that under specifically controlled conditions, the deamidase treated wheat gluten (byproduct from food industry), which contain a large amount of amidated amino acids, could be a useful procedure for improving their functional properties.

Knowledge Management: Journal