Characterization and bioassay of penicillin (Penicillium Chrysogenum) produced from oranges (Citrus Sinensis) collected from Na'ibawa orange market, Kano State, Nigeria

ABSTRACTThis study aimed at producing penicillin from the rotten, fresh and preserved rotten oranges. The samples were prepared by extracting the orange juice and their pHs adjusted to pH ۶.۵ using ۱% calcium hydroxide; then sterilized for fifteen minutes using an autoclave at ۱۲۱°C. The samples were then inoculated with Penicillium chrysogenum and kept for fifteen days. The inoculated samples were analysed using Thin Layer Chromatography (TLC), Fourier Transform Infrared Spectroscopy (FTIR), and Gas Chromatography Mass Spectrometry (GC-MS). The FTIR spectra of rotten and fresh oranges were similar to that of the Benzathine benzylepenicillin standard. The spectra indicated a broad band at ۳۳۴۲ cm-۱, representing bonded –OH stretching vibration and the –NH stretching of a protein. The peaks appearing in the ۲۹۷۷ cm-۱ and ۲۸۳۶ cm-۱ regions may be attributed to the asymmetric and symmetric stretching vibration of -CH۲, respectively. The strong adsorption band at ۱۶۵۹ cm-۱ represents -C=O stretching vibration and NH deformation (amide I). The peak at ۱۵۶۶ cm-۱ was assigned to a motion combining both –NH bending (amide II) and –CN stretching vibration of the protein. The typical amide III band, which represents -COO- anions, appeared at ۱۳۸۰ cm-۱. The weak peak at ۱۳۹۸ cm-۱ was induced by the -C-N stretching vibration of amine groups. The strong adsorption peaks ۱۲۴۹ cm-۱ and ۱۰۲۶ cm-۱ are indicators of –SO۳ groups and the -C-N stretching vibration of amine groups, respectively. The moderate peak at ۸۸۸ cm-۱ may be attributed to aromatic –CH stretching vibration. The fingerprint zone of the spectra, ranging from ۶۵۰ cm-۱ to ۸۰۰ cm-۱, represents phosphate or sulphur functional groups. Bioactive compounds were identified from the penicillin produced from rotten and fresh oranges using GC-MS analysis. The most prevalent were; cis-۱۳-octadecenoic acid, methyl ester, hexadecanoic acid, pentyl ester, ۹-octadecenoic acid (Z)-,۲,۳-dihydroxypropylester, ۹-Octadecenoic acid (Z)-,۲-hydroxy-۱-(hydroxyl methyl)ethyl ester and ۴-Oxatricyclo [۲۰.۸.۰.۰(۷,۱۶)] triaconta-۱(۲۰),۷(۱۶)-diene, at a retention times of; ۱۳.۶ min, ۱۶.۰۵ min, ۲۱.۹۰ min, ۲۱.۹۴ min and ۳۰.۸۶ min respectively. The highest zone of inhibitions observed before recovery due to Escherichia coli and Streptococcus for the rotten and fresh oranges were; ۲۲.۴ mm and ۱۸.۲۴ mm respectively; while inhibition was not observed for the preserved rotten oranges. However, after recovery, the highest zone of inhibitions observed due to E.coli, Methicillin-resistant Staphylococcus aureus (MRSA), Meticillinic-Sencitive Staphylococcus aureus (MSSA) and Klepsiella were ۲۶ mm for the standard, ۸ mm for the fresh orange, ۳۲ mm for the fresh orange, and ۳۴ mm for the rotten orange respectively. The highest yield (۴۰۰ mg/cm۳) of the produced penicillin was the recorded for the rotten orange before optimization. The yield remains the same after optimization. The study demonstrated that fresh and rotten oranges are viable substrates for the fermentation media to be used in cultivating fungus for the production of natural penicillin antibiotics.

کلیدواژه ها:

Antibiotics ، production ، Spectroscopy ، Waste Management