Utilization of agro-industrial by-products for the production of kojic acid

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dc.contributor.advisorSahota, Param Pal
dc.contributor.authorSharma, Neha
dc.date.accessioned2021-10-26T04:57:26Z
dc.date.available2021-10-26T04:57:26Z
dc.date.issued2021
dc.description.abstractThe present study involves a novel fermentative bioprocess optimized for the production of kojic acid (2-hydroxymethyl-5-hydroxy-γ-pyrone) to combat the problem of oxidative stress posed by its chemically synthesized forms. Four promising kojic acid producing fungal strains (KJPAU1, KJPAU2, KJPAU3, KJPAU4) were screened with the Rapid Bio Oxidation (RaBiO) method (91.3 % isolation rate) with the yield of 40.13, 21.1, 9.20, 14.6 gL-1 respectively on GYE medium. The isolates showed tolerance to the wide range of temperature (20-50 °C), pH (3.5-6.5), assimilated Carbohydrates (61.9 %), utilized Nitrogen sources (66.67 %), induced hydrolysis of starch, cellulose. Based on 18SrRNA sequencing, atoxicogenic KJPAU1, KJPAU2 isolates were characterized as Aspergillus nomius (MH752445), A. oryzae (EF568052) respectively with a >98 % similarity values. Among the agro-industrial substrates (Kinnow residue, Amla waste, Sugarcane bagasse, Wheat straw) studied for kojic acid production after 10 days of fermentation with the inoculation rate @ 20 % having the final concentration of 3×105 spores/mL with intermittent shaking conditions, the parameters of optimized fermentative bioprocess consisted of Kinnow waste residue (20 %), LCUM (4 %), MgSO4 (0.1 %), KCl (0.05 %), pH 6.0, temperature 30 °C with the maximum production of 11.64 gL-1. The kojic acid productivity in Kinnow waste was found to be higher, 66.7 %, 16.6 % and 52.1 % from Wheat straw, Amla pomace, Sugarcane bagasse residues respectively. The maximum activity of Glucose dehydrogenase (44.3 µmol TPF h-1) at optimized production conditions was estimated on tenth day at pH (6.0), temperature (30 °C), KCl (0.05 %). The downstream processing of kojic acid with the direct concentration and crystallization method showed >42.23 % recovery with the 97 % purity. The TLC solvent system, Toluene: Ethyl Acetate: Formic Acid= 3:6:1 facilitated the separation at Rf value (0.64), detected under UV lamp (336 nm). The purity of fermentatively produced kojic acid was confirmed by C18 RP-HPLC with the Acetonitrile: Phosphate buffer (50:50) as the mobile phase at a flow rate 0.3 mL/ min, with the retention time of 9.143 min (approx.) at 254 nm. With the FTIR spectrum for functional groups at 3270.8 cm-1, 3179.43 cm-1 (- OH), 2925.17 cm-1, 2854.05 cm-1 (aliphatic- CH), 1660.59 cm-1 (cyclic-C=O), 1611.11 cm-1 (C=C), 1472.61 cm-1 (deformation of-CH2), 1074.04 cm-1 (cyclic C-O-C), 943.58 cm-1 , 863.66 cm-1 and 775.65 cm-1 (1, 4 α -disubstituted ring) confirmed the crystalline purity (97 %) of kojic acid. To prevent pericarp browning of litchi fruit, 8mM kojic acid application for five minutes delayed melanosis up to nine days. The results of oral fed kojic acid dosage @ 1, 2.5, 5 % in albino mice for 28 days showed no adverse effects on body weight (g), food/ water intake, general behavior.en_US
dc.identifier.citationSharma,Neha (2021). Utilization of agro-industrial by-products for the production of kojic acid (Unpublished Ph.D. Dissertation). Punjab Agricultural University, Ludhiana, Punjab, India.en_US
dc.identifier.urihttps://krishikosh.egranth.ac.in/handle/1/5810177273
dc.keywordsKojic acid, Aspergillus sp.s, Agro-industrial residues, Glucose dehydrogenase, Crystallization, Mice model, Pericarp browningen_US
dc.language.isoEnglishen_US
dc.pages105en_US
dc.publisherPunjab Agricultural University, Ludhianaen_US
dc.research.problemUtilization of agro-industrial by-products for the production of kojic aciden_US
dc.subMicrobiologyen_US
dc.themeUtilization of agro-industrial by-products for the production of kojic aciden_US
dc.these.typePh.Den_US
dc.titleUtilization of agro-industrial by-products for the production of kojic aciden_US
dc.typeThesisen_US
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