Heterologous Expression and Characterization of a pH-Stable Chitinase from <i>Micromonospora aurantiaca</i> with a Potential Application in Chitin Degradation
Han-Zhong Guo,
Dou Wang,
Hui-Ting Yang,
Yu-Le Wu,
Yong-Cheng Li,
Guang-Hua Xia,
Xue-Ying Zhang
Affiliations
Han-Zhong Guo
Hainan Engineering Research Center of Aquatic Resources Efficient Utilization in South China Sea, Key Laboratory of Food Nutrition and Functional Food of Hainan Province, Key Laboratory of Seafood Processing of Haikou, National R&D Branch Center for Prawn Processing Technology (Haikou), School of Food Science and Engineering, Hainan University, Haikou 570228, China
Dou Wang
Hainan Engineering Research Center of Aquatic Resources Efficient Utilization in South China Sea, Key Laboratory of Food Nutrition and Functional Food of Hainan Province, Key Laboratory of Seafood Processing of Haikou, National R&D Branch Center for Prawn Processing Technology (Haikou), School of Food Science and Engineering, Hainan University, Haikou 570228, China
Hui-Ting Yang
Hainan Engineering Research Center of Aquatic Resources Efficient Utilization in South China Sea, Key Laboratory of Food Nutrition and Functional Food of Hainan Province, Key Laboratory of Seafood Processing of Haikou, National R&D Branch Center for Prawn Processing Technology (Haikou), School of Food Science and Engineering, Hainan University, Haikou 570228, China
Yu-Le Wu
Hainan Engineering Research Center of Aquatic Resources Efficient Utilization in South China Sea, Key Laboratory of Food Nutrition and Functional Food of Hainan Province, Key Laboratory of Seafood Processing of Haikou, National R&D Branch Center for Prawn Processing Technology (Haikou), School of Food Science and Engineering, Hainan University, Haikou 570228, China
Yong-Cheng Li
Hainan Engineering Research Center of Aquatic Resources Efficient Utilization in South China Sea, Key Laboratory of Food Nutrition and Functional Food of Hainan Province, Key Laboratory of Seafood Processing of Haikou, National R&D Branch Center for Prawn Processing Technology (Haikou), School of Food Science and Engineering, Hainan University, Haikou 570228, China
Guang-Hua Xia
Hainan Engineering Research Center of Aquatic Resources Efficient Utilization in South China Sea, Key Laboratory of Food Nutrition and Functional Food of Hainan Province, Key Laboratory of Seafood Processing of Haikou, National R&D Branch Center for Prawn Processing Technology (Haikou), School of Food Science and Engineering, Hainan University, Haikou 570228, China
Xue-Ying Zhang
Hainan Engineering Research Center of Aquatic Resources Efficient Utilization in South China Sea, Key Laboratory of Food Nutrition and Functional Food of Hainan Province, Key Laboratory of Seafood Processing of Haikou, National R&D Branch Center for Prawn Processing Technology (Haikou), School of Food Science and Engineering, Hainan University, Haikou 570228, China
To promote the bioconversion of marine chitin waste into value-added products, we expressed a novel pH-stable Micromonospora aurantiaca-derived chitinase, MaChi1, in Escherichia coli and subsequently purified, characterized, and evaluated it for its chitin-converting capacity. Our results indicated that MaChi1 is of the glycoside hydrolase (GH) family 18 with a molecular weight of approximately 57 kDa, consisting of a GH18 catalytic domain and a cellulose-binding domain. We recorded its optimal activity at pH 5.0 and 55 °C. It exhibited excellent stability in a wide pH range of 3.0–10.0. Mg2+ (5 mM), and dithiothreitol (10 mM) significantly promoted MaChi1 activity. MaChi1 exhibited broad substrate specificity and hydrolyzed chitin, chitosan, cellulose, soluble starch, and N-acetyl chitooligosaccharides with polymerization degrees ranging from three to six. Moreover, MaChi1 exhibited an endo-type cleavage pattern, and it could efficiently convert colloidal chitin into N-acetyl-D-glucosamine (GlcNAc) and (GlcNAc)2 with yields of 227.2 and 505.9 mg/g chitin, respectively. Its high chitin-degrading capacity and exceptional pH tolerance makes it a promising tool with potential applications in chitin waste treatment and bioactive oligosaccharide production.
