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. 2023 Apr 27;15(9):2079.
doi: 10.3390/polym15092079.

Crude Enzyme Concentrate of Filamentous Fungus Hydrolyzed Chitosan to Obtain Oligomers of Different Sizes

Cleidiane Gonçalves E Gonçalves  1 , Lúcia de Fátima Henriques Lourenço  2 , Hellen Kempfer Philippsen  3 , Alberdan Silva Santos  4 , Lucely Nogueira Dos Santos  1 , Nelson Rosa Ferreira  1   2
Affiliations

Crude Enzyme Concentrate of Filamentous Fungus Hydrolyzed Chitosan to Obtain Oligomers of Different Sizes

Cleidiane Gonçalves E Gonçalves et al. Polymers (Basel). .

Abstract

Chitosan is a non-cytotoxic polysaccharide that, upon hydrolysis, releases oligomers of different sizes that may have antioxidant, antimicrobial activity and the inhibition of cancer cell growth, among other applications. It is, therefore, a hydrolysis process with great biotechnological relevance. Thus, this study aims to use a crude enzyme concentrate (CEC) produced by a filamentous fungus to obtain oligomers with different molecular weights. The microorganism was cultivated in a liquid medium (modified Czapeck-with carboxymethylcellulose as enzyme inducer). The enzymes present in the CEC were identified by LC-MS/MS, with an emphasis on cellobiohydrolase (E.C 3.2.1.91). The fungus of the Aspergillus genus was identified by amplifying the ITS1-5.8S-ITS2 rDNA region and metaproteomic analysis, where the excreted enzymes were identified with sequence coverage greater than 84% to A. nidulans. Chitosan hydrolysis assays compared the CEC with the commercial enzyme (Celluclast 1.5 L®). The ability to reduce the initial molecular mass of chitosan by 47.80, 75.24, and 93.26% after 2.0, 5.0, and 24 h of reaction, respectively, was observed. FTIR analyses revealed lower absorbance of chitosan oligomers' spectral signals, and their crystallinity was reduced after 3 h of hydrolysis. Based on these results, we can conclude that the crude enzyme concentrate showed a significant technological potential for obtaining chitosan oligomers of different sizes.

Keywords: chitosan; enzyme; filamentous fungus; hydrolysis.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Dendogram based on peptides built by Unipept (https://unipept.ugent.be/, accessed on 11 December 2022). Phylogenetic tree based on the list of peptides identified using the lowest common ancestor (LCA) method.
Figure 2
Figure 2
Enzyme activity profile (CMCase) in different culture media for Aspergillus nidulans: ■—modified czapek culture medium (Czm); ●—medium cellulose peptone yeast extract (CPY).
Figure 3
Figure 3
Chitosan hydrolysis profile to obtain oligomers as a function of reaction time and different enzyme concentrates: ■—CE (commercial enzyme); ●—CEC (crude enzyme concentrate).
Figure 4
Figure 4
Chitosan hydrolysis as a function of reaction time using A. nidulans crude enzyme concentrate.
Figure 5
Figure 5
FT-IR spectrum of chitosan hydrolyzed with crude enzyme concentrate at times of 0 h, 1 h, 3 h, 5 h, and 24 h.
See this image and copyright information in PMC

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