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Changes in Biochemical Characteristics and Activities of Ripening Associated Enzymes in Mango Fruit During the Storage at Different Temperatures

Md. Anowar Hossain
Department of Biochemistry and Molecular Biology, University of Rajshahi, Rajshahi 6205, Bangladesh

Md. Masud Rana
Department of Biochemistry and Molecular Biology, University of Rajshahi, Rajshahi 6205, Bangladesh

Yoshinobu Kimura
Department of Biofunctional Chemistry, Graduate School of Natural Science and Technology, Okayama University, Okayama 700-8530, Japan

Hairul Azman Roslan
Genetic Engineering Laboratory, Department of Molecular Biology, Faculty of Resource Science and Technology,Universiti Malaysia Sarawak, 94300 Kota Samarahan, Sarawak, Malaysia

Abstract/ Executive Summary:

As a part of the study to explore the possible strategy for enhancing the shelf life of mango fruits, it investigated the changes in biochemical parameters and activities of ripening associated enzymes of Ashwina hybrid mangoes at 4-day regular intervals during storage at −10°C, 4°C, and 30 ± 1°C. Titratable acidity, vitamin C, starch content, and reducing sugar were higher at unripe state and gradually decreased with the increasing of storage time at all storage temperatures while phenol content, total soluble solid, total sugar, and nonreducing sugar contents gradually increased. The activities of amylase, α-mannosidase, α-glucosidase, and invertase increased sharply within first few days and decreased significantly in the later stage of ripening at 30 ± 1°C. Meanwhile polyphenol oxidase, β-galactosidase, and β-hexosaminidase predominantly increased significantly with the increasing days of storage till later stage of ripening. At −10°C and 4°C, the enzymes as well as carbohydrate contents of storage mango changed slightly up to 4 days and thereafter the enzyme became fully dormant. The results indicated that increase in storage temperature and time correlated with changes in biochemical parameters and activities of glycosidases suggested the suppression of β-galactosidase and β-hexosaminidase might enhance the shelf life of mango fruits.

Keywords: Biochemical characteristics, Mango, Storage, Different temperatures

Location: Protein Research Laboratory, Department of Biochemistry and Molecular Biology, University of Rajshahi, Bangladesh.

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Mango

Objectives:

The aim of the study was to investigate the changes in biochemical characteristics and activities of various glycosidases of Ashwina hybrid mangoes at 4-day regular intervals during storage at -10°C, 4°C, and 30⁰ ± 1°C for an identification of biochemical parameters responsible for ripening process.

Methodology:

The experiments were carried out using the equipment available in the Protein Research Laboratory, Department of Biochemistry and Molecular Biology, University of Rajshahi, Bangladesh. Unless stated otherwise, all the reagents, synthetic and other substrates, used for performing the experiments were from Sigma-Aldrich and Carl-Roth GmbH, Germany. Collection of Samples and Storage Procedure: Fresh mature green mangoes (Ashwina hybrid variety) were collected from Banesshar Bazar at Rajshahi, Bangladesh. Samples were sorted at the same maturity level and the pathogen-infected and/or mechanical damaged mangoes were discarded. The mangoes were stored at −10°C, 4°C, and 30 ± 1°C (room temperature) for a period of 12 days. Three mangoes were taken out from each group of samples on 4 days of regular interval of time such as on 0th, 4th, 8th, and 12th day and kept in −80°C for further analysis. Preparation of Crude Extract: Fruits were cut into small pieces and homogenized by adding equal volume of extraction buffer (100 mM Tris-HCl, pH 7.0 with 0.25 M NaCl and 4 mM PMSF). The homogenized fruit was kept overnight at 4°C. After passing through four layers of muslin cloth, the extract was centrifuged at 10,000 ×g for 15 min to separate the debris. Supernatant was collected as crude extract. The various biochemical parameters were determined according to the following protocols. Total Soluble Solids (TSS) and Titratable Acidity (TA): TSS was determined according to the method as described by Mazumdar and Majumder using digital bench refractometer (range 0–32%). TA was determined according to the method as described by Hortwitz. Ten milliliters of crude extract was taken in a beaker and titrated with 0.1 N NaOH after adding 2-3 drops of phenolphthalein as an indicator. Vitamin C: Approximately 2 to 3 g of mango flesh was cut into small pieces and homogenized well with 20 mL of 3% metaphosphoric acid and filtered through double layers of muslin cloth. The filtrate was centrifuged at 3,000 ×g for 10 min and the clear supernatant was titrated with 2,6-dichlorophenol indophenol solution. The amount of vitamin C in the extract was determined by comparing with the titration curve of standard vitamin C solution. Result was expressed in mg/100 g of fresh fruit. Starch Content: The starch content of the mango flesh was determined by the anthrone method. Two grams of mango was cut into small pieces and homogenized well with 20 mL water. Twice the volume of ethanol was added to the filtrate to precipitate the polysaccharide, mainly starch. After keeping overnight at 4°C, the precipitate was collected by centrifugation at 3,000 ×g for 15 min. The precipitate was heated to dryness and dissolved in 40 mL of 1 M HCl and then heated at 70⁰C for few minutes. It was then transferred to a volumetric flask and diluted to 100 mL with 1 M HCl. Aliquot of 1 mL of the extract of each sample was pipetted out into test tubes and 4 mL of the anthrone reagent was added to the each test tube and mixed well. The tubes were placed in a boiling water bath for 10 min and cooled. The amount of starch present in mango flesh was calculated from the standard curve of different concentrations of glucose and expressed as g/100 g of fresh fruits. Total Sugar (TS), Reducing Sugar (RS), Non reducing Sugar (NRS), and Phenol Content (PC): Two grams of mango flesh was cut into small pieces and plunged into 20 mL of boiling ethanol for 5–10 min. After cooling, it was crushed thoroughly in a mortar with a pestle. The homogenate was filtered through two layers of muslin cloth and refiltered through a Whatman no. 41 filter paper. The extract was evaporated to dryness over a steam bath and subsequently cooled. The residues were dissolved in 100 mL of distilled water and the resulting solution was used as flesh extract (sample stock) for the estimation of TS, RS, and PC. 2.6.1. TS TS content of mango was determined calorimetrically by the anthrone method. The test tubes were placed in a boiling water bath for 10 minutes. After cooling, the absorbencies was measured at 680 nm against a blank reagent. The total sugar present in the sample tubes was determined from the standard curve prepared using different concentrations of glucose and expressed as g/100 g of mango flesh. RS Reducing sugar content of the mango was determined by dinitrosalicylic acid (DNS) method. Aliquot of 3 mL of the flesh extract was pipetted out into three test tubes and 3 mL of DNS reagent was added to each solution and mixed well. The test tubes were heated for 5 min in a boiling water bath. After developing the color, 1 mL of 40% Rochelle salt was added when the content of the tubes was still warm. The test tubes were then cooled under running tap water. The absorbencies of the solution was measured at 575 nm in a colorimeter against a blank reagent.

Source of Information: Biomed Res Int. 2014; 2014: 232969. Published online 2014 Jul 22. doi: 10.1155/2014/232969

Article Link (Online Journal): http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4129145/

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The results indicated that increase in storage temperature and time correlated with changes in biochemical parameters and activities of glycosidases suggested the suppression of β-galactosidase and β-hexosaminidase might enhance the shelf life of mango fruits.

Knowledge Management: Journal