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Biosynthesis and extrusion of β-chitin nanofibers by diatoms
Published in Antonio Trincone, Enzymatic Technologies for Marine Polysaccharides, 2019
Chitinases promote the degradation of chitin by hydrolytic cleavage of the β-1,4 linkage, and have two primary categories: (1) endochitinases (EC 3.2.1.14), which cleave internal linkages, primarily down to chitobiose (N,N’-diacetylchitobiose); and (2) β-1,4-N-acetylglucosaminidases (EC 3.2.1.30), which cleave soluble chitin oligomers (chitobiose, chitotriose, and chitotetraose) at the nonreducing end into N-acetylglucosamine monomer (Patil et al. 2000). Cyclotella cryptica has a nearly complete chitin degradation pathway with 22 chitinases (Traller et al. 2016), and T. pseudonana has at least 20 genes that encode putative chitin-degrading chitinases (Durkin et al. 2009). Most of the chitinases are shared within the Thalassiosirales group and have similarity to either bacterial or fungal chitinases.
Chaperone-assisted soluble expression and characterization of chitinase chiZJ408 in Escherichia coli BL21 and the chitin degradation by recombinant enzyme
Published in Preparative Biochemistry & Biotechnology, 2022
Ping Yu, Xinxin Wang, Jian Ma, Qili Zhang, Qingwei Chen
In the previous study, a strain Aeromonas sp. ZJ408 was isolated in our laboratory. This strain could secret a low-yield high molecular weight chitinase chiZJ408. In this study, in order to enhance the soluble expression of this chitinase in E. coli cells, the Aeromonas sp. ZJ408 chitinase gene and E. coli K12 molecular chaperone GroELS gene were cloned and co-expressed in E. coli BL21 by constructing a recombinant plasmid pETDuet-1-chiZJ408-GroELS. The expression of the molecular chaperone GroELS significantly promoted the correct folding of high molecular weight chitinase chiZJ408, and therefore improved its soluble expression in E. coli BL21. Moreover, chitin degradation products by chitinase chiZJ408 were determined by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). Interestingly, only N,N’-diacetyl-chitobiose (GlcNAc)2 was produced by chitin degradation via chitinase chiZJ408.