Biochemical Properties of a Cold-Active Chitinase from Marine Trichoderma gamsii R1 and Its Application to Preparation of Chitin Oligosaccharides

Abstract

The enzymatic degradation of different chitin polymers into chitin oligosaccharides (COSs) is of great significance given their better solubility and various biological applications. Chitinase plays a pivotal role in the enzymatic preparation of COSs. Herein, a cold-adapted and efficient chitinase (ChiTg) from the marine Trichoderma gamsii R1 was purified and characterized. The optimal temperature of ChiTg was 40 °C, and the relative activity at 5 °C was above 40.1%. Meanwhile, ChiTg was active and stable from pH 4.0 to 7.0. As an endo-type chitinase, ChiTg exhibited the highest activity with colloidal chitin, then with ball-milled and powdery chitin. In addition, ChiTg showed high efficiency when hydrolyzing colloidal chitin at different temperatures, and the end products were mainly composed of COSs with one to three degrees of polymerization. Furthermore, the results of bioinformatics analysis revealed that ChiTg belongs to the GH18 family, and its acidic surface and the flexible structure of its catalytic site may contribute to its high activity in cold conditions. The results of this study provide a cold-active and efficient chitinase and ideas for its application regarding the preparation of COSs from colloidal chitin.

Strategies to Prepare Chitin and Chitosan-Based Bioactive Structures Aided by Deep Eutectic Solvents: A Review

“…Therefore, research efforts have been made to improve the efficacy of enzymatic processing of chitin and chitosan. One strategy would involve the synergistic utilization of physical treatments, such as microwave [188], ultrasonication [189], or mechanochemistry [190][191][192][193], to help in the enzymatic degradation of chitin by promoting its partial solubilization. Another option would be the use of strong organic solvents which have been reported to dissolve chitin [57], such as such as LiCl/dimethylacetamide (DMAc) [194], CaCl2/MeOH [195], or hexafluoroisopropanol (HFIP, [196]).…”

Heterologous Expression and Characterization of a pH-Stable Chitinase from Micromonospora aurantiaca with a Potential Application in Chitin Degradation

“…However, the low-commercial-value marine biomass produced also has a negative effect on the environment and ecosystem when not utilized effectively. Every year, approximately 9 × 10 9 tons of crustaceans including crabs, shrimps, and lobsters are generated, with roughly 60% being disposed of as solid waste due to their inedibility [ 1 , 2 ]. This crustacean waste, containing 15–40% chitin [ 3 ], presents a cheap and renewable resource.…”

“…Several bacterial chitinases have been identified for their degradation activities toward chitinous substrates, such as the recombinant chitinase Chisb from Bacillus sp. DAU101, which hydrolyzed colloidal chitin into a mixture of (GlcNAc) 1-5 [ 19 ]; wild-type chitinase ChiTg from Trichoderma gamsii R1, which converted colloidal chitin into (GlcNAc) 1-3 [ 2 ]; and the recombinant chitinase CcCti1 from Corallococcus sp. EGB, which could degrade colloidal chitin into (GlcNAc) 6 as its major product [ 20 ].…”

Thermal Stability Enhancement of L-Asparaginase from Corynebacterium glutamicum Based on a Semi-Rational Design and Its Effect on Acrylamide Mitigation Capacity in Biscuits

“…However, this increased flexibility also leads to a decrease in the stability of these enzymes. As temperature rises, even reaching room temperature, cold-active enzymes become sensitive to heat, resulting in poor storage stability and making it difficult for them to meet industrial requirements [25][26][27]. Therefore, a thermostable enzyme holds significant and valuable potential for application in the food industry.…”