Immobilization of Cellulolytic Enzymes in Accurel® MP1000

• Published in: Reactions (Basel, Switzerland) Vol. 4
• Main Authors: Baruque, J. R. S., Carniel, A., Sales, J. C. S., Ribeiro, B. D., Do Nascimento, R. P., Itabaiana Junior, Ivaldo
• Format: Journal Article
• Language: English
• Published: MDPI 14.01.2024
• Series: Reactions
• Subjects: Chemical Sciences; Trichoderma harzianum; cellulases; cellulase immobilization; Accurel (R) MP1000; Celluclast (R) 1.5 L
• ISSN: 2624-781X; 2624-781X
• DOI: 10.3390/reactions4020019

Cellulases are a class of enzymes of great industrial interest that present several strategic applications. However, the high cost of enzyme production, coupled with the instabilities and complexities of proteins required for hydrolytic processes, still limits their use in several protocols. Therefore, enzyme immobilization may be an essential tool to overcome these issues. The present work aimed to evaluate the immobilization of cellulolytic enzymes of the commercial enzyme cocktail Celluclast® 1.5 L in comparison to the cellulolytic enzyme cocktail produced from the wild strain Trichoderma harzianum I14-12 in Accurel® MP1000. Among the variables studied were temperature at 40 °C, ionic strength of 50 mM, and 72 h of immobilization, with 15 m·L −1 of proteins generated biocatalysts with high immobilization efficiencies (87% for ACC-Celluclast biocatalyst and 95% for ACC-ThI1412 biocatalyst), high retention of activity, and specific activities in the support for CMCase (DNS method), FPase (filter paper method) and β-glucosidase (p-nitrophenyl-β-D-glucopyranoside method). Presenting a lower protein concentration (0.32 m·L−1) than the commercial Celluclast® 1.5 L preparation (45 m·L−1), the ACC-ThI1412-derived immobilized biocatalyst showed thermal stability at temperatures higher than 60 °C, maintaining more than 90% of the residual activities of FPase, CMCase, and β-glucosidase. In contrast, the commercial-free enzyme presented a maximum catalytic activity at only 40 °C. Moreover, the difference in molecular weight between the component enzymes of the extract was responsible for different hydrophobic and lodging interactions of proteins on the support, generating a robust and competitive biocatalyst.