Subramani, A.K.Ramachandra, R.Thote, S.Govindaraj, V.Vanzara, P.Raval, R.Raval, K.2026-02-042024International Journal of Biological Macromolecules, 2024, 264, , pp. -1418130https://doi.org/10.1016/j.ijbiomac.2024.130499https://idr.nitk.ac.in/handle/123456789/21197Chitin, an abundant polysaccharide in India, is primary by-product of the seafood industry. Efficiently converting chitin into valuable products is crucial. Chitinase, transforms chitin into chitin oligomers, holds significant industrial potential. However, the crystalline and insoluble nature of chitin makes the conversion process challenging. In this study, a recombinant chitinase from marine bacteria Bacillus aryabhattai was developed. This enzyme exhibits activity against insoluble chitin substrates, chitin powder and flakes. The chitinase gene was cloned into the pET 23a plasmid and transformed into E. coli Rosetta pLysS. IPTG induction was employed to express chitinase, and purification using Ni-NTA affinity chromatography. Optimal chitinase activity against colloidal chitin was observed in Tris buffer at pH 8, temperature 55°C, with the presence of 400 mM sodium chloride. Enzyme kinetics studies revealed a V<inf>max</inf> of 2000 μmole min−1 and a K<inf>m</inf> of 4.6 mg mL−1. The highest chitinase activity against insoluble chitin powder and flakes reached 875 U mg−1 and 625 U mg−1, respectively. The chitinase demonstrated inhibition of Candida albicans, Fusarium solani, and Penicillium chrysogenum growth. Thin Layer Chromatography (TLC) and LC-MS analysis confirmed the production of chitin oligomers, chitin trimer, tetramer, pentamer, and hexamer, from chitin powder and flakes using recombinant chitinase. © 2024 Elsevier B.V.Affinity chromatographyBacteriologyChitinCloningEnzyme activityEnzyme kineticsEscherichia coliOligomersSodium chlorideThin layer chromatographyChitin oligomerChitinase activitiesChitinase genesChitinasesConversion processCrystalline chitinsE. coliIndustrial potentialsInsoluble chitin substrateMarine bacteriumSubstrateschitinchitinaseoligomersodium chloridetetrameraffinity chromatographyantifungal activityArticleBacillusBacillus aryabhattaibacterium isolationCandida albicansDNA determinationdrug injection volumeenzyme activityenzyme kineticsfungus growthFusarium solaniliquid chromatography-mass spectrometrymarine bacteriumnonhumanparticle sizePenicillium chrysogenumpHpolyacrylamide gel electrophoresisprotein purificationSanger sequencingtemperaturethin layer chromatographyturnover numberultra performance liquid chromatographychemistrygeneticspowderHydrogen-Ion ConcentrationPowders