Chitin is a ubiquitous structural biopolymer with significant potential for various high-value applications. Chitin deacetylases (CDAs) can potentially deacetylate chitinous molecules, producing chitosans and partially acetylated chitooligosaccharides (COS). Here, we describe the expression and characterization of a CDA from the filamentous fungus Absidia coerulea, AcCDA1, active toward colloidal chitin, partially deacetylated chitin, chitosan with different degrees of acetylation, and COS. When incubated with COS with degrees of polymerization 3-6, the enzyme rapidly produced fully deacetylated COS, which is a relatively rare property among known CDAs. The mode of action of AcCDA1 toward penta-N-acetyl chitopentaose ((GlcNAc)5) was studied by mass spectrometry, using a novel approach for labeling the reducing N-acetyl-D-glucosamine unit of the COS with chitooligosaccharide oxidase (FgChitO) from Fusarium graminearum. Kinetic studies with COS revealed high enzyme efficiency with kcat/Km values reaching 18.8 mM-1 s-1 for (GlcNAc)5. The enzyme removed 12% of the acetyl groups in colloidal chitin whereas it drastically reduced the degree of acetylation of various chitosans. A comparative study with three other CDAs showed that AcCDA1 is highly efficient enzyme and revealed both similarities and differences. These findings show that various CDAs can achieve different deacetylation targets during the enzymatic processing of chitinous materials.
Laura Barahona-Pérez, María Martínez-Ranz, Zarah Forsberg et al.· Carbohydrate Polymers· 0 citations
Abstract With this status report, we aim to provide a timely snapshot of the protein engineering field as a broad and rapidly advancing discipline that integrates computational, molecular biology, structure-guided, evolutionary, and synthetic approaches to create new and improved proteins with tailored structures and useful functions. The report is organized into eight thematic areas spanning core methodologies and major application domains, including enzymes, therapeutics, detection, synthetic biology, and materials. Contributions from experts across these areas highlight both the historical foundations and recent advances in their respective fields, with particular emphasis on the growing influence of machine learning and artificial intelligence-based methods. Emerging from this broad overview is a central message: protein engineering appears to be entering a golden age, defined by a rapidly accelerating pace of progress, even as significant challenges in design, screening, and real-world application remain. Looking ahead, the continued integration of computational and experimental strategies is poised to further accelerate the impact of protein engineering across an expanding range of economically and societally important sectors, from therapeutics and molecular imaging to diagnostics, plastic recycling, and industrial chemistry.
Hui-wang Ai, Frances H. Arnold, Doug Barrick et al.· Protein engineering, design...· 0 citations
It is demonstrated that these multidomain LPMOs from food-borne intestinal disease-causing Gram-positive bacteria from the genera Bacillus and Listeria contain conserved, yet structurally distinct copper(I)-binding motifs on their non-catalytic third domain.
Eirik G. Kommedal, Hanne Berggreen, Synnøve Elisa Rønnekleiv et al.· bioRxiv· 0 citations
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