食品科学 ›› 2025, Vol. 46 ›› Issue (18): 73-74.doi: 10.7506/spkx1002-6630-20250207-019

• 生物工程 • 上一篇    下一篇

卢氏发光杆菌κ-卡拉胶酶Cgk483的重组表达及其酶解产物的体外降脂功能

陈菁,潘旭思,黄浩阳,刘浩,汪卓,赵巧丽,戴梓茹,钟赛意   

  1. (1.广东海洋大学深圳研究院,广东?深圳 518108;2.广东海洋大学食品科技学院,广东省水产品加工与安全重点实验室,广东省海洋生物制品工程实验室,广东省海洋食品工程技术研究中心,广东?湛江 524088;3.北部湾大学食品工程学院,广西?钦州 535011)
  • 出版日期:2025-09-25 发布日期:2025-08-19
  • 基金资助:
    深圳市科技重大专项(KCXFZ20240903094014019);深圳市国际合作项目(GJHZ20240218114715029); “十三五”国家重点研发计划重点专项(2020YFD0901101);广东省高校创新团队项目(2021KCXTD021)

Recombinant Expression of κ-Carrageenase Cgk483 from Photobacterium rosenbergii and in Vitro Lipid-lowering Activity of κ-Carrageenan Oligosaccharides Prepared Using It

CHEN Jing, PAN Xusi, HUANG Haoyang, LIU Hao, WANG Zhuo, ZHAO Qiaoli, DAI Ziru, ZHONG Saiyi   

  1. (1. Shenzhen Institute of Guangdong Ocean University, Shenzhen 518108, China; 2. Guangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Guangdong Province Engineering Laboratory for Marine Biological Products, Guangdong Provincial Engineering Technology Research Center of Seafood, College of Food Science and Technology, Guangdong Ocean University, Zhanjiang 524088, China; 3. College of Food Engineering, Beibu Gulf University, Qinzhou 535011, China)
  • Online:2025-09-25 Published:2025-08-19

摘要: 本研究旨在探讨卢氏发光杆菌(Photobacterium rosenbergii)中κ-卡拉胶酶Cgk483的重组表达及其在卡拉胶寡糖制备中的应用,并对得到的卡拉胶寡糖进行结构表征以及体外降脂效果分析。通过基因克隆和异源表达,成功获得电泳纯的重组酶Cgk483,其分子质量为35.89 kDa,纯化后的酶活力提升10 倍,达到489.88 U/mg。酶学性质研究表明,重组酶Cgk483的最适反应温度和pH值分别为40 ℃和8.0,在40 ℃以下和pH 5.0~10.0范围内具有较好稳定性,Fe2+、Na+和Ba2+能提升重组酶Cgk483的活性。利用重组酶Cgk483成功制备得到κ-卡拉胶寡糖,由新卡拉二糖、新卡拉四糖和新卡拉六糖组成。结构分析显示酶解未破坏其主链,但使其羟基等基团暴露,提高了κ-卡拉胶寡糖的溶解性。体外降脂研究表明,κ-卡拉胶寡糖能显著抑制氧化型低密度脂蛋白引起的RAW264.7巨噬细胞泡沫化,减少脂质蓄积。本研究制备了高效制备κ-卡拉胶寡糖的工具酶,并揭示了其酶解产物在降脂方面的应用潜力,可为κ-卡拉胶寡糖的高效制备及动脉粥样硬化防治拓展新思路。

关键词: κ-卡拉胶酶;卡拉胶寡糖;降脂;卢氏发光杆菌

Abstract: This study aimed to explore the recombinant expression of κ-carrageenase Cgk483 from Photobacterium rosenbergii and its application in preparing carrageenan oligosaccharides, which were structurally characterized and evaluated for in vitro hypolipidemic effects. Through gene cloning and heterologous expression, the electrophoretically pure recombinant enzyme Cgk483 was successfully obtained, with a molecular mass of 35.89 kDa and its specific activity was 489.88 U/mg, which was approximately 10 times higher than that of the wild-type enzyme. Enzyme kinetics studies showed that the optimal reaction temperature and pH for Cgk483 were 40 ℃ and 8.0, respectively. It was stable at temperatures below 40 ℃ and within the pH range of 5.0–10.0. Fe2+, Na+, and Ba2+ were found to enhance the enzyme activity. Using the recombinant enzyme Cgk483, κ-carrageenan oligosaccharides consisting of disaccharide, tetrasaccharide and hexasaccharide were successfully prepared. Structural analysis indicated that the enzymatic hydrolysis did not disrupt the main chain but resulted in the exposure of hydroxyl and other groups, improving the solubility of κ-carrageenan oligosaccharides. In vitro hypolipidemic assay demonstrated that κ-carrageenan oligosaccharides significantly inhibited oxidized low-density lipoprotein (Ox-LDL)-induced foam cell formation in RAW264.7 macrophages, reducing lipid accumulation. This study provides a new idea for the efficient preparation of κ-carrageenan oligosaccharides for use in the prevention and treatment of atherosclerosis.

Key words: κ-carrageenase; κ-carrageenan oligosaccharide; lipid-lowering; Photobacterium rosenbergii

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