Purification and characterization of a novel β-1,3-glucanase from Arca inflata and its immune-enhancing effects

Chunlei Li; Yao Wen; Ying He; Jianhua Zhu; Xin Yin; Jianing Yang; Lixin Zhang; Liyan Song; Xuekui Xia; Rongmin Yu

doi:10.1016/j.foodchem.2019.03.131

Purification and characterization of a novel β-1,3-glucanase from Arca inflata and its immune-enhancing effects

Food Chem. 2019 Aug 30:290:1-9. doi: 10.1016/j.foodchem.2019.03.131. Epub 2019 Mar 26.

Authors

Chunlei Li¹, Yao Wen², Ying He³, Jianhua Zhu³, Xin Yin⁴, Jianing Yang³, Lixin Zhang⁵, Liyan Song³, Xuekui Xia⁶, Rongmin Yu⁷

Affiliations

¹ Biotechnological Institute of Chinese Materia Medica, Jinan University, 601 Huangpu Avenue West, Guangzhou 510632, China; Biology Institute, Qilu University of Technology (Shandong Academy of Sciences), Jinan 250013, China.
² Biotechnological Institute of Chinese Materia Medica, Jinan University, 601 Huangpu Avenue West, Guangzhou 510632, China.
³ Department of Natural Medicinal Chemistry, College of Pharmacy, Jinan University, Guangzhou 510632, China.
⁴ Biology Institute, Qilu University of Technology (Shandong Academy of Sciences), Jinan 250013, China.
⁵ Biology Institute, Qilu University of Technology (Shandong Academy of Sciences), Jinan 250013, China; State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China. Electronic address: lxzhang@ecust.edu.cn.
⁶ Biology Institute, Qilu University of Technology (Shandong Academy of Sciences), Jinan 250013, China. Electronic address: xiaxk@sdas.org.
⁷ Biotechnological Institute of Chinese Materia Medica, Jinan University, 601 Huangpu Avenue West, Guangzhou 510632, China; Department of Natural Medicinal Chemistry, College of Pharmacy, Jinan University, Guangzhou 510632, China. Electronic address: tyrm@jnu.edu.cn.

PMID: 31000023
DOI: 10.1016/j.foodchem.2019.03.131

Abstract

A novel β-1,3-glucanase from Arca inflata was purified using chromatography methods. It was determined as a glycoprotein comprising 23.65% carbohydrate content with O-linked glycan and showed specific activity of 90.01 ± 1.2 U/mg against laminarin. The optimal pH and temperature for the activity of the glucanase were 6.0 and 40 °C, respectively. The affinity parameter of the glucanase using laminarin was determined as K_d = 13.09 μM. The activity of the glucanase was 27 ± 2.6% enhanced by 2-mM Mn²⁺ ions and inhibited by 40-50% using 2-mM Zn²⁺, Cu²⁺, or Ba²⁺ ions. The glucanase showed an endo-type cleavage mode and hydrolyzed laminarin into glucoses, disaccharides, trioligosaccharides, and tetraoligosaccharides. Otherwise, the glucanase exhibited immune-enhancing effects via significantly increasing the phagocytic activity of macrophages and inducing the release of nitric oxide, tumor necrosis factor α, and interleukin-6 in RAW264.7 cells. It might be used as a bifunctional additive for the food industry.

Keywords: Arca inflata; Biochemical characterization; Endo-β-1,3-Glucanase; Glycoprotein; Immune-enhancing activity.

MeSH terms

Animals
Bivalvia / enzymology*
Glucan 1,3-beta-Glucosidase / isolation & purification*
Glucan 1,3-beta-Glucosidase / metabolism
Glucan 1,3-beta-Glucosidase / pharmacology*
Glucans / metabolism
Hydrogen-Ion Concentration
Hydrolysis
Immunologic Factors / isolation & purification*
Immunologic Factors / metabolism
Immunologic Factors / pharmacology*
Macrophages / drug effects
Macrophages / immunology
Macrophages / metabolism
Mice
RAW 264.7 Cells
Substrate Specificity
Temperature

Substances

Glucans
Immunologic Factors
laminaran
Glucan 1,3-beta-Glucosidase