Hidrólise da quitosana: obtenção de um extrato enzimático e caracterização do produto hidrolisado

Abstract

Chitin, extensively found in crustaceans exoskeletons, insects, and microorganisms, has limited usage due to its low solubility in aqueous solution, requiring its partial deacetylation to obtain chitosan. Chitosan's depolymerization has attracted considerable attention, as its oligomers have high water solubility, biocompatibility, and non-toxicity, as well as beneficial properties such as antimicrobial, antioxidant, and antitumor properties. For this reason, in this research, a review article was produced (Chapter I) based on the main methods of chitosan hydrolysis, besides analyzing the parameters that influence the acquisition, and characteristics of hydrolysis results, effectively and at a lower cost. Among the approaches studied, enzymatic hydrolysis excels due to its control ease and performance under milder conditions, making it possible to use low-cost enzymes belonging to the glycoside hydrolases group. Thus, enzymatic hydrolysis was defined as a technique for various sizes of chitosan acquisition (Chapter II) through the production of an enzymatic extract (integral enzymatic extract - IEE) from a filamentous fungus strain. The enzyme identification present in the IEE showed exo-chitinases, endo-chitinase, and cellobiohydrolase. Considering the same reaction conditions, the IEE showed greater efficiency than the commercial enzyme (Celluclast 1.5 L®), which was used as a parameter because it is an enzyme capable of cleaving the β-1,4-glycosidic bond of chitosan - similar to chitosanase, besides presenting a lower cost. The IEE reduced the molecular weight of chitosan by 47.80; 75.24 and 93.26% at 2.0; 5.0, and 24 h, respectively. Through the FTIR analysis, a lower absorbance of the spectral signals of chitosan oligomers was detected, and the crystallinity reduced after 3.0 h of hydrolysis. Based on this study, we can infer that enzymatic hydrolysis, under established conditions, is effective at obtaining lower molecular weight chitosan using unpurified crude extract.