Biochemical Purification and Characterization of a Truncated Acidic, Thermostable Chitinase from Marine Fungus for N-acetylglucosamine Production

Overview

Journal Front Bioeng Biotechnol

Date 2022 Oct 21

PMID 36267443

Authors

Bin He

Liyan Yang

Dengfeng Yang

Minguo Jiang

Chengjin Ling

Hailan Chen

Feng Ji

Lixia Pan

Affiliations

Soon will be listed here.

Abstract

N-acetylglucosamine (GlcNAc) is widely used in nutritional supplement and is generally produced from chitin using chitinases. While most GlcNAc is produced from colloidal chitin, it is essential that chitinases be acidic enzymes. Herein, we characterized an acidic, highly salinity tolerance and thermostable chitinase ChiJ, identified from the marine fungus df673. Using AlphaFold2 structural prediction, a truncated Δ30ChiJ was heterologously expressed in and successfully purified. It was also found that it is active in colloidal chitin, with an optimal temperature of 45°C, an optimal pH of 4.0, and an optimal salt concentration of 3% NaCl. Below 45°C, it was sound over a wide pH range of 2.0-6.0 and maintained high activity (≥97.96%) in 1-7% NaCl. A notable increase in chitinase activity was observed of Δ30ChiJ by the addition of Mg, Ba, urea, and chloroform. ChiJ first decomposed colloidal chitin to generate mainly N-acetyl chitobioase, which was successively converted to its monomer GlcNAc. This indicated that ChiJ is a bifunctional enzyme, composed of chitobiosidase and β-N-acetylglucosaminidase. Our result suggested that ChiJ likely has the potential to convert chitin-containing biomass into high-value added GlcNAc.

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