Phylogenetic Classification of Chitinases in Cotton Bollworm (Helicoverpa armigera), Expression and Analysis of Enzymatic Characterization on Group Ⅶ Chitinase
ABUDUNASIER Maimaitiailii, LI Meng-Ge, BAO Jing, SUI Hui, LIU Xiao-Ning*
Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046, China
Abstract Insect chitinases are mainly involved in important physiological growth processes such as molting, degradation of peritrophic membranes, cell proliferation and immune defense. In this study, seven chitinases from Helicoverpa armigera and other insect chitinases sequences were downloaded from NCBI. Their domains and constructed phylogenetic tree were analyzed. The results showed that 6 of chitinases belonged to group Ⅰ, Ⅱ, Ⅲ, Ⅳ, Ⅶ and Ⅷ chitinase, respectively, and 1 chitinase belonged to the new group. Group Ⅶ chitinase (HaCHTⅦ) was rarely reported, It was cloned in this study. Fusion protein His-HaCHTⅦ was expressed in Escherichia coli transetta (DE3) and purified. The degradation activity of fusion protein against colloidal chitin and N-acetylated chitobiose was determined. Results showed that His-HaCHTⅦ had no degradative activity on colloidal chitin, and had the degradative activity on N-acetylchitobiose. The optimum reaction pH value and temperature condition was at 7 and 25℃, respectively. The kinetic parameter of michaelis constant (Km) and maximum speed (Vmax) were 0.486 2 mmol/L and 2.715 1 µg/(mL·min), respectively. Therefore, HaCHTⅦ was probably to degrade oligosaccharides to N-acetylglucosamine in H. armigera and then provided substrate to synthesize the new chitin component in the development and growth. Present findings may lead us to a better understand of the function of the group Ⅶ chitinase from H. armigera and may provide useful information for pest control.
ABUDUNASIER Maimaitiailii,LI Meng-Ge,BAO Jing, et al. Phylogenetic Classification of Chitinases in Cotton Bollworm (Helicoverpa armigera), Expression and Analysis of Enzymatic Characterization on Group Ⅶ Chitinase[J]. 农业生物技术学报, 2019, 27(3): 495-503.
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