A jacalin-related lectin domain-containing lipase from chestnut (Castanea crenata): Purification, characterization, and protein identification

Jun Heo; Chang Woo Kwon; Juno Lee; Haena Park; Hyunjong Yu; Pahn-Shick Chang

doi:10.1016/j.crfs.2022.10.033

A jacalin-related lectin domain-containing lipase from chestnut (Castanea crenata): Purification, characterization, and protein identification

Curr Res Food Sci. 2022 Nov 2:5:2081-2093. doi: 10.1016/j.crfs.2022.10.033. eCollection 2022.

Authors

Jun Heo¹, Chang Woo Kwon², Juno Lee¹, Haena Park¹, Hyunjong Yu^{2

3}, Pahn-Shick Chang^{1

2

4

3}

Affiliations

¹ Department of Agricultural Biotechnology, Seoul National University, Seoul, 08826, Republic of Korea.
² Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, 08826, Republic of Korea.
³ Center for Agricultural Microorganism and Enzyme, Seoul National University, Seoul, 08826, Republic of Korea.
⁴ Center for Food and Bioconvergence, Seoul National University, Seoul, 08826, Republic of Korea.

Abstract

A novel lipase (triacylglycerol acylhydrolase, EC 3.1.1.3) was discovered from Korean chestnut (Castanea crenata). The lipase was isolated and purified by ammonium sulfate precipitation and a fast protein liquid chromatography system equipped with HiTrap DEAE-Sepharose Fast Flow, HiTrap Q-Sepharose Fast Flow, and HiPrep Sephacryl S-100 Hi-Resolution columns. The purified C. crenata lipase showed a 15.8% yield, purification fold number of 465.8, and specific activity against triolein of 88.5 mU/mg. The enzyme exhibited hydrolytic activity toward tributyrin, trilaurin, and triolein, and was maximally active at pH 8.0 and 35 °C, with triolein used as the substrate. The activation energy (E_a) and deactivation energy (E_d) of triolein hydrolysis were 38.41 and 83.35 kJ/mol, respectively. In the enzyme kinetic study, V_max, K_m, and k _cat were 110.58 mU/mg, 0.11 mM, and 0.221 min^-1, respectively. The relatively low K_m value indicated that the lipase has high affinity for its substrate. Moreover, Mg²⁺ and Ca²⁺ increased the lipase activity to 115.4% and 108.3%, respectively. The results of peptide fingerprinting revealed that the C. crenata lipase with a molecular weight of 33.3 kDa was structurally similar to the mannose-binding lectin of the jacalin-related lectin domain superfamily, implying that it has potential as a therapeutic agent for use in the biomedical industry.

Keywords: Castanea crenata; Characterization; Chestnut lipase; Jacalin-related lectin domain; Protein identification; Purification.