Biochemical characterization of two β-N-acetylglucosaminidases from Streptomyces violascens for efficient production of N-acetyl-D-glucosamine

Chitin, one of the most abundant renewable biopolymers on Earth, is commercially available from crustacean wastes. One critical step in converting chitin to high-value products is its degradation by chitinolytic enzymes to N-acetyl-D-glucosamine (GlcNAc), which plays a significant role in functional food and pharmaceutical industries. Here, we cloned and biochemically characterized two novel beta-N-acetylglucosaminidases named SvNag2557 (family-84) and SvNag4755 (family-3) from Streptomyces violascens ATCC 27968. Both SvNag2557 and SvNag4755 exhibited strict substrate specificity toward N-acetyl chitooligosaccharides with GlcNAc as the sole product. Thus, a one-pot production for pure GlcNAc from chitin by an enzyme cocktail reaction was further developed. Under the co-action of an endo-type chitinase SaChiA4 and SvNag2557 (mass ratio 1:2), the final conversion rates of colloidal chitin and ionic liquid pretreated chitin to GlcNAc were 80.2% and 73.8% with GlcNAc purities of 99.7% and 96.8%, respectively.

Automated summary

Chitin is converted to GlcNAc by chitinases, essential for food and pharmaceutical industries. Two novel beta-N-acetylglucosaminidases were cloned and characterized, enabling the one-pot production of pure GlcNAc from chitin through enzymatic reactions.