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Enzyme Activities and Mobilization of Nutrients in Brassica (brassica Spp.) and Wheat (triticum Aestivum L.) Seeds During Germination

N K Sana
Department of Chemistry, University of Rajshahi, Rajshahi-6205, Bangladesh

B C Sarkar
Department of Chemistry, University of Rajshahi, Rajshahi-6205, Bangladesh

M A K Azad
Department of Chemistry, University of Rajshahi, Rajshahi-6205, Bangladesh

M E Huque
Department of Biochemistry & Molecular Biology,University of Rajshahi, Rajshahi-6205, Bangladesh

R K Shaha
Department of Chemistry, University of Rajshahi, Rajshahi-6205, Bangladesh

Abstract/ Executive Summary:

Context: Determination of the activities of hydrolytic enzymes from germinating wheat seeds and investigation of degraded nutrients from brassica and wheat seeds at different periods of germination are important factors for identification of richest sources of hydrolytic enzymes and nutrients. Objectives: To study the activities of hydrolytic enzymes and degradation of seed storage substances of brassica (Brassica napus L.) and wheat (Triticum aestivum L.) seeds during germination. Materials and Methods: Three varieties of brassica (Brassica napus L., B. juncea L. and B. campestris L.) and three varieties of wheat (Akbar, Kanchan and Agrani) seeds were analyzed. Amylase, invertase, protease and lipase activities were assayed. Degradation of seed storage nutrients during germination were determined by conventional biochemical methods. Results: The activities of amylase, invertase, protease and lipase varied from 8.02 – 48.69, 2.45 – 15.32, 20.96 – 45.45 and 2.72 – 12.76 units/ ml respectively. Degradation of nutrients in the three species of brassica and wheat seeds was also studied at different periods of germination. The amount of free sugar in brassica and wheat seeds ranged from 0.93 – 4.27% and 3.82 – 4.88%; reducing sugar content from 0.012 - 0.093% and 0.032 - 0.078%; starch from 1.42 – 4.70% and 10.26 – 69.65%; total protein from 3.1 – 25.37% and 2.3 – 18.37%; water-soluble protein from 1.5 – 14.24% and 1.0 – 6.50%; and oil content from 2.49 – 43.6% and 1.04 – 1.92% respectively. Conclusion: The results suggest that the extracts from brassica and wheat seeds can be good sources of nutrients and hydrolytic enzymes which are applicable in food industry to improve food quality.

Keywords: Brassica, Wheat, Carbohydrate, Protein, Lipid, Hydrolytic Enzyme.

Location: Department of Chemistry, University of Rajshahi

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Wheat, Mustard

Objectives:

To study the activities of hydrolytic enzymes and degradation of seed storage substances of brassica (Brassica napus L.) and wheat (Triticum aestivum L.) seeds during germination.

Methodology:

Materials: Three species of brassica seeds (Brassica napus L., B. juncea L. and B. campestris L.) and three varieties of wheat (Akbar, Kanchan and Agrani) seeds were collected from Bangladesh Agriculture Research Institute Regional Centres of Pabna, Dinajpur, Rajshahi and Jessore. After collection, the seeds were cleaned, dried in the sunlight and kept in a polyethylene bag and stored in the deep freeze (-10°C) for further biochemical analysis. Glucose, BSA, Dinitrosalicylic acid (DNS) and Sodium tungstate were purchased from Sigma Chemicals Ltd., USA. Chloroform, Ethanol and Trichloro acetic acid (TCA) were purchased from Pharmacia Fine Chemicals Ltd., Sweden. All other chemicals used were commercially available and of high purity. Germination of seed: Quality and mature seeds of brassica and wheat were soaked in distilled water within a glass beaker with potassium permanganate for six hours for avoiding the growth of microorganism on seed surfaces during germination. The seeds were taken out from water and scattered in 20 cm petri dishes on two sheets of Whatman No. 1 filter paper moistened with distilled water. The dishes were then placed in the light room at 25 ºC for 120 h including soaking time. The germinating seeds at different hours (0, 24, 32, 48, 96 and 120 h) were collected and stored separately in the deep freeze for further experimental purposes. Preparation of the crude extract: Germinated seeds (100g) were crushed in a mortar and suspended in 40 ml of 30% acetone. After occasional gentle stirring for 3 h at 4°C the suspension was filtered through double layer of muslin cloth. Filtrate was centrifuged in a refrigerated centrifuge at 5500 rpm for 15 min at 4ºC. The supernatant was used as “crude extract”. The crude extract was saturated to 30 - 50% by the addition of solid ammonium sulphate under constant and gentle stirring at 4ºC. The resulting precipitate was collected by centrifugation, dissolved in minimum volume of pre-cooled distilled water and dialyzed against distilled water for 24 h at 4ºC. The dialyzed solution was then centrifuged in a refrigerated centrifuge machine at 5500 rpm for 15 min. The clear supernatant thus obtained was designated as “crude enzyme solution”. Measurement of enzyme activity: Amylase activity was assayed following the method as described by Jayaraman (1985). Starch solution (1%) was used as substrate (1 g in 100 ml of 0.1 M phosphate buffer, pH 6.7). Invertase activity was assayed following the method of Mahadevan and Sridhar (1982). Sucrose solution (2.5%) was used as a substrate. The protease activity was measured following the method of Kunitz (1947). The milk protein casein was used as substrate. Lipase activity was assayed by the method described by Sugihara et al. (1990). Olive oil was used as substrate.

Source of Information: J. bio-sci. 17: 101-106, 2009, ISSN 1023-8654

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The results suggest that the extracts from brassica and wheat seeds can be good sources of nutrients and hydrolytic enzymes which are applicable in food industry to improve food quality.

Knowledge Management: Journal