Purification and characterization of cutinase from Botrytis cinerea and effect on blueberry cuticle
2023
Meng Liu | Zihao Zong | Xiangjun Fang | Ruiling Liu | Honglei Mu | Hangjun Chen | Ben Niu | Haiyan Gao
An extracellular cutinase in Botrytis cinerea was isolated and its biochemical properties and effects on the penetration of pathogens were evaluated. This cutinase had a specific activity of 6.03 U/μg of protein after 22.33-fold purification, and yielded a recovery of 50.5%. The optimum pH and temperature for the enzymatic activity of cutinase were 9 and 45 °C, respectively. The cutinase was stable between 30 and 80 °C and pH 4.0–10.0. Fe3+ and Cu2+ strongly increased cutinase activity while Mn2+ completely inhibited it. The enzyme could maintain its activity in the presence of methanol, ethanol and isopropanol, but was significantly inhibited in acetonitrile, n-hexane and acetone. Kinetic parameters KM, Vmax and kcat were 0.21 mM, 303 μmol•min−1•mg−1 protein and 303.03 s−1 respectively. The purified cutinase could also destroy blueberry wax crystal and decrease the cuticle, indicating a positive role on the penetration of pathogens for blueberry fruits. This study provides useful information for fruit postharvest preservation.
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