Enzymatic hydrolysates of κ-carrageenan by κ-carrageenase-CLEA immobilized on amine-modified ZIF-8 confer hypolipidemic activity in HepG2 cells

Chunhua Zhu; Mingjing Mou; Leilei Yang; Zedong Jiang; Mingjing Zheng; Zhipeng Li; Tao Hong; Hui Ni; Qingbiao Li; Yuanfan Yang; Yanbing Zhu

doi:10.1016/j.ijbiomac.2023.126401

Enzymatic hydrolysates of κ-carrageenan by κ-carrageenase-CLEA immobilized on amine-modified ZIF-8 confer hypolipidemic activity in HepG2 cells

Int J Biol Macromol. 2023 Dec 1:252:126401. doi: 10.1016/j.ijbiomac.2023.126401. Epub 2023 Aug 18.

Authors

Chunhua Zhu¹, Mingjing Mou¹, Leilei Yang¹, Zedong Jiang², Mingjing Zheng², Zhipeng Li², Tao Hong², Hui Ni³, Qingbiao Li², Yuanfan Yang⁴, Yanbing Zhu⁵

Affiliations

¹ College of Ocean Food and Biological Engineering, Jimei University, Xiamen 361021, China.
² College of Ocean Food and Biological Engineering, Jimei University, Xiamen 361021, China; Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering, Xiamen 361021, China; Research Center of Food Biotechnology of Xiamen City, Xiamen 361021, China.
³ College of Ocean Food and Biological Engineering, Jimei University, Xiamen 361021, China; Xiamen Ocean Vocational College, Xiamen 361102, China.
⁴ College of Ocean Food and Biological Engineering, Jimei University, Xiamen 361021, China; Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering, Xiamen 361021, China; Research Center of Food Biotechnology of Xiamen City, Xiamen 361021, China. Electronic address: yuanfan@jmu.edu.cn.
⁵ College of Ocean Food and Biological Engineering, Jimei University, Xiamen 361021, China; Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering, Xiamen 361021, China; Research Center of Food Biotechnology of Xiamen City, Xiamen 361021, China. Electronic address: yanbingzhu@jmu.edu.cn.

PMID: 37597638
DOI: 10.1016/j.ijbiomac.2023.126401

Abstract

κ-Carrageenase can degrade κ-carrageenan to produce bioactive κ-carrageenan oligosaccharides (KCOs) that have potential applications in pharmaceutical, food, agricultural, and cosmetics industries. Immobilized enzymes gain their popularity due to their good reusability, enhanced stability, and tunability. In this study, the previously characterized catalytic domain of Pseudoalteromonas purpurea κ-carrageenase was covalently immobilized on the synthesized amine-modified zeolitic imidazolate framework-8 nanoparticles with the formation of cross-linked enzyme aggregates, and the immobilized κ-carrageenase was further characterized. The immobilized κ-carrageenase demonstrated excellent pH stability and good reusability, and exhibited higher optimal reaction temperature, better thermostability, and extended storage stability compared with the free enzyme. The KCOs produced by the immobilized κ-carrageenase could significantly decrease the TC, TG, and LDL-C levels in HepG2 cells, increase the HDL-C level in HepG2 cells, and reduce the free fatty acids level in Caco-2 cells. Biochemical assays showed that the KCOs could activate AMPK activity, increase the ratios of p-AMPK/AMPK and p-ACC/ACC, and downregulate the expression of the lipid metabolism related proteins including SREBP1 and HMGCR in the hyperlipidemic HepG2 cells. This study provides a novel and effective method for immobilization of κ-carrageenase, and the KCOs produced by the immobilized enzyme could be a potential therapeutic agent to prevent hyperlipidemia.

Keywords: Hypolipidemic activity; Immobilized κ-carrageenase; κ-Carrageenan oligosaccharides.

MeSH terms

AMP-Activated Protein Kinases*
Bacterial Proteins* / chemistry
Caco-2 Cells
Carrageenan / chemistry
Enzymes, Immobilized
Glycoside Hydrolases / chemistry
Hep G2 Cells
Humans
Oligosaccharides / chemistry

Substances

Carrageenan
AMP-Activated Protein Kinases
Bacterial Proteins
Glycoside Hydrolases
Oligosaccharides
Enzymes, Immobilized