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Comparative Antibacterial and Antioxidant Activities of Edible and Non-edible Parts of Unripe Banana (musasapientum L.) Fruit

Md. Shoriful Islam
Department of Agricultural Botany, Sher-e-Bangla Agricultural University, Dhaka-1207, Bangladesh

Ferdousi Begum
Department of Agricultural Botany, Sher-e-Bangla Agricultural University, Dhaka-1207, Bangladesh

Farzana Ashrafi Neela
Department of Agricultural Botany, Sher-e-Bangla Agricultural University, Dhaka-1207, Bangladesh

Mohammad Firoz Alam
Department of Agricultural Botany, Sher-e-Bangla Agricultural University, Dhaka-1207, Bangladesh

Abstract/ Executive Summary:

Comparative antibacterial and antioxidant in vitroactivities of edible (pulp) and non-edible (peel) parts of unripe banana fruit were investigated. Peels and pulps of fruit were extracted using methanol and acetone separately. Antibacterial property of these extracts was evaluated against four species of multiple drug resistant (MDR) bacteria namely Escherichia coli, Klebsiella sp, Salmonella sp. and Shigellasp.using disc diffusion technique. When compared between pulp and peel on the basis of zone of inhibition created by them, pulp and peel exhibited 14.5 mm and 16.2 mm, respectively, whereas the standard drug, Kanamycine (30μg/disc) showed >30mm. In case of MIC, pulp was ranged from 200-300 mg/ml, while peel was from 200-250 mg/ml. As like MIC, MBC also found better in peel (400-550 mg/ml) than pulp (450-600 mg/ml). Results showed that non-edible part was better than edible part for controlling bacteria. Antioxidant activity of the extracts was evaluated by total phenolic content determination and 1, 1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay. DPPH scavenging activities of non-edible part (70.62%) was found higher than edible part (54.79%), while 100% was observed by ascorbic acid at same concentration (100μg/ml). In case of phenol content, edible part of banana showed lower phenolic content (71.49 mgL-1GAE / g dry material) than non-edible part (92.76 mgL-1GAE / g dry material). Comparing between pulp and peel, peel found better than pulp in every case, e.g., antibacterial (both MIC and MBC), antioxidant activities and total phenol content. The results suggest that banana peels could serve as potential source of bioactive compounds and can be utilized effectively without being wasted.

Keywords: Musasp., Antibacterial activities, Antioxidant activities, phenol content.

Location: Plant Taxonomist, Department of Botany, University of Rajshahi, Bangladesh

Start Date:

End Date:

Program Area: Knowledge Management

Commodity: Banana

Objectives:

The objective was comparative in vitro studies on antibacterial and antioxidant activities between edible (pulp) and not-edible (peel) solvent extracts of unripe banana fruit available in local market.

Methodology:

Plant material Banana (Musa sapientum) fruits were collected from Rajshahi local market, Saheb Bazar, at green stage confirm no chemical treatment and the sample was identified and authenticated as Musa sapientum (L.) by Dr. AHM Mahbubur Rahman, Professor and Plant Taxonomist, Department of Botany, University of Rajshahi, Bangladesh and a voucher specimen was deposited at the Herbarium of the department. The freshly collected bananas were washed with distilled water to remove dirt, followed by separation of edible (whole fruit –peel = pulp) and non-edible (whole fruit –pulp = peel) parts using autoclaved knife. Then the edible and non-edible parts were chopped separately into pieces and dried at room temperature (32-35°C) for five days until a constant weight was obtained. 250g of each of the parts were coarsely powdered using a mortar and pestle, and were further reduced to powder using an electric blender. Then the powdery materials (edible and non-edible) were stored separately in air tight bottles for further use. Extraction procedures Plant extracts were prepared using the methods of Sultana et al. (2009) with slight modification. Each of the powdered air-dried plant material was extracted with methanol and acetone solvent separately. 5g of each powdered sample was mixed in a conical flask with 100ml of organic solvent, and then allowed to soak at ambient temperature for 24 h. The extracts were then filtered with tetron cloth into a beaker and then using Whatman no. 1 filter paper. The filtrates concentrated at 40°C using a rotary evaporator (EYELA, SB-651, Rikakikai Co. Ltd. Tokyo, Japan). Then the concentrated extracts were collected in a screw cap tube and stored at 4°C for further use. The yield was found to be 2.45±0.87 and 4.66±1.63% w/w in methanol and acetone, with reference to the air dried powdery plant material. The dried extracts were dissolved in dimethylsulphoxide and subjected to antibacterial activity. Test organisms Four species of multiple drug resistant (MDR) bacteria were used for this investigation namely Escherichia coli, Klebsiella sp, Salmonella sp. and Shigella sp. All clinical isolates obtained from the Plant Biotechnology and Microbiology Laboratory of the Department of Botany, Rajshahi University. The organisms were periodically subcultured and maintained in nutrient agar slant at 4 0C. Antibacterial activities Antibacterial activity of the methanolic and acetonic extracts of the plant sample (edible and non-edible part of unripe banana) was evaluated using disc diffusion method according to Bauer et al. (1966) with slight modifications. For determination of antibacterial activity, bacterial cultures were adjusted to 0.5 McFarland turbidity standard and inoculated onto Nutrient agar (oxoid) plates (diameter: 15cm). Sterile filter paper discs (diameter 6mm) impregnated with 100µl of extract dilutions reconstituted in minimum amount of solvent at concentrations of 50 and 100mg/ml were applied over each of the culture plates previously seeded with the 0.5 McFarland and 106cfu/ml cultures of bacteria. After placing the disc, the plate was inverted and incubated at 37°C for 24 hours. Then the plate was examined and measured the diameters of the clear zones on the agar surface including the diameter of the disk. The zones were measure to the nearest millimeter using a ruler and values <8 mm were considered as not active against microorganisms (Bhalodia and Shukla, 2011). The experiment was replicated three times to confirm the reproducible results. Commercially available kanamycin discs (30 µg/disc) were used as positive control while the discs prepared using the appropriate solvents only served as negative control. Determination of minimum inhibitory concentration and minimum bactericidal concentration The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined according to Doughari et al. (2007) with slide modification. The MIC of each extracts was determined against each of the test bacteria in varying concentrations of 100, 200, 300, 400, 500, 600 and 700 mg/ml. Then the nutrient broth (9ml) tube was added with 1ml of test organism previously diluted to 0.5 McFarland turbidity standards. A tube containing only nutrient broth was seeded with the test bacteria, as described above, to serve as controls. All the tubes were then incubated at 37°C for 24 h and then examined for growth by observing for turbidity. MIC was measured by the concentration at which there was no change of turbidity (Ajaiyeoba et al., 2003). Determination of antioxidant activity using free radical scavenging activity (DPPH) The 1, 1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging by the extracts was determined by the method described by Braca et al. (2001). 200 µl plant extract of different strength (5, 25, 50, 100, 500 µg/ml) was mixed with 2 ml of a 0.004% methanol solution of DPPH. After 30 min, the absorbance was determined at 517 nm using a UV spectrophotometer (Shimadzu, UV-1601PC) against a blank. Absorbance of DPPH solution only without the extract or standard agent was used as control. The percentage scavenging activity of the extracts was calculated using the formula: % scavenging activity = {(A0 – A1) /A0} × 100; where A0 is the absorbance of the control and A1 is the absorbance of the extract or standard. Determination of IC50 value A percent inhibition versus concentration curve was plotted and the concentration of sample required for 50 percent inhibition was determined and expressed as IC50 value. The lower the IC50 value indicates high antioxidant capacity. The effective concentration of sample required to scavenge DPPH radical by 50% (IC50 value) was obtained by linear regression analysis of dose-response curve plotting between % inhibition and concentrations. Determination of total phenolic content Estimation of total phenol content was conducted using the Folin–Ciocalteu (F–C) reagent spectrophotometric method described by Ainsworth and Gillespie (2007) with slight modification. The F– C assay relies on the transfer of electrons in alkaline medium from phenolic compounds to phosphomolybdic or phosphotungstic acid complexes, which are determined spectroscopically at 765nm. Although the electrons transfer reaction is not specific for phenolic compound. For creating the calibration curve, gallic acid was used as a standard material.

Source of Information: International Journal of Biosciences, Vol. 16, No. 3, p. 290-298, 2020

Article Link (Online Journal): http://dx.doi.org/10.12692/ijb/16.3.290-298

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

In the present study,an attempt has been made to determine the antimicrobial and antioxidant properties of edible (pulp) and non-edible (peel) parts of unripe banana fruit in vitro. All the methanolic and acetonic extracts of pulp and peel have been found to show significant antibacterial activities against used MDR bacteria. Thus extracts from the plant can be used to control infections caused by Escherichia coli, Klebsiella sp., Shigella sp and Salmonella sp.Results also showed that pulp and peel extracts contain high amount of antioxidants along with phenolic compounds. Comparing between pulp and peel, in case of antibacterial and antioxidant activities, peel found better than pulp. The results suggest that parts of fruit like peels could serve as potential source of bioactive compounds and can be utilized effectively without being wasted. The study also suggests that the pulp and peel of banana may be used for the extraction and preparation of various antioxidant and antibacterial formulations in pharmaceutical industries.

Knowledge Management: Journal