Abstract
A novel chitinase gene of 888 bp from Streptomyces bacillaris was cloned and expressed in Escherichia coli BL21. The purified recombinant enzyme (SbChiAJ103) was identified as the first microbial-derived family 19 endochitinase that showed exochitinase activity. SbChiAJ103 exhibited the substrate preference for N-acetylchitooligosaccharides with even degrees of polymerization and the capability to specifically hydrolyze colloidal chitin into (GlcNAc)2. Mono-methyl adipate was employed as a novel linker for the efficient covalent immobilization of chitinase on magnetic nanoparticles (MNPs). The immobilized SbChiAJ103, SbChiAJ103@MNPs, exhibited superior pH tolerance, temperature stability, and storage stability than free SbChiAJ103. Even after incubation at 45 °C for 24 h, SbChiAJ103@MNPs could retain more than 60.0% initial activity. As a result, the enzymatic hydrolysis yield of SbChiAJ103@MNPs increased to 1.58 times that of free SbChiAJ103. Moreover, SbChiAJ103@MNPs could be reused by convenient magnetic separation. After 10 recycles, SbChiAJ103@MNPs could retain almost 80.0% of its initial activity. The immobilization of the novel chitinase SbChiAJ103 paves the way to the efficient and eco-friendly commercial production of (GlcNAc)2.
Key points
• The first microbial GH19 endochitinase with exochitinase activity was reported.
• Mono-methyl adipate was first employed to immobilize chitinase.
• SbChiAJ103@MNPs showed excellent pH stability, thermal stability, and reusability.
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This work was supported by the earmarked fund for CARS-48.
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F. S., C. X, and X. M. conceived and designed research. A. X. performed research. Y. H. contributed analytical tools. A. X. and Y. H. analyzed data. A. X. and Y. H. wrote the manuscript. All authors read and approved the manuscript.
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Xing, A., Hu, Y., Wang, W. et al. A novel microbial-derived family 19 endochitinase with exochitinase activity and its immobilization. Appl Microbiol Biotechnol 107, 3565–3578 (2023). https://doi.org/10.1007/s00253-023-12523-2
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DOI: https://doi.org/10.1007/s00253-023-12523-2