生物学杂志

生物学杂志 ›› 2025, Vol. 42 ›› Issue (1): 55-.doi: 10.3969/j.issn.2095-1736.2025.01.055

• 研究报告 • 上一篇    下一篇

新琼四糖的分离纯化及其保湿活性研究

刘　骁1,2， 杨子宜2,3， 吴超城2,3， 吕佰霖2,3， 曾润颖2,3   

  1. 1. 福州大学 先进制造学院， 晋江 362251； 2. 自然资源部第三海洋研究所 海洋生物资源开发利用
    工程技术创新中心， 厦门 361005； 3. 福建省海岛资源生态监测与保护利用重点实验室， 平潭 350400
  • 出版日期:2025-02-18 发布日期:2025-02-12
  • 通讯作者: 曾润颖，博士，教授，研究方向为海洋资源应用，E-mail:zeng@tio.org.cn
  • 作者简介:刘骁，硕士，研究方向为海洋资源应用，E-mail:liuxiao_lyris@126.com
  • 基金资助:
    中央引导的地方科技发展资助项目（2022L3022）； 福建省科技资助项目（2022H0036， 2023N0034）

Neoagarotetraose: isolation, purification and moisturizing activity

LIU Xiao1,2, YANG Ziyi2,3, WU Chaocheng2,3, LYU Bailin2,3, ZENG Runying2,3   

  1. 1. School of Advanced Manufacturing, Fuzhou University, Jinjiang 362251, China; 2. Marine Biological Resources
    Development and Utilization Engineering Technology Innovation Center, Third Institute of Oceanography, MNR,
    Xiamen 361005, China; 3. Fujian Key Laboratory of Island Monitoring and Ecological Development
    (Island Research Center, MNR), Pingtan 350400, China
  • Online:2025-02-18 Published:2025-02-12

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摘要： 采用凝胶过滤色谱法对重组琼胶酶AgaP4383降解琼胶的终产物新琼寡糖进行分离纯化，并采用离子交换色谱（IEC）和电喷雾飞行时间质谱（ESI-TOF-MS）对纯化产物的纯度及分子量进行分析；利用CCK-8法检测新琼四糖对角质形成细胞活力的影响，并通过酶联免疫吸附试验和免疫荧光法检测新琼四糖对角质形成细胞保湿相关蛋白（FLG、AQP3、ZO-1）表达量的影响。结果表明：重组琼胶酶AgaP4383降解琼胶得到的产物经过Bio-Gel P2 fine凝胶柱分离可得到纯度为96.53%、相对分子量为630的新琼四糖，得率为78.4%；新琼四糖在0.0625~1 mg/mL的质量浓度范围内对角质形成细胞的活力无明显影响，生物相容性良好；新琼四糖在质量浓度为0.125 mg/mL和0.5 mg/mL时能够显著提高角质形成细胞中丝聚蛋白FLG、水通道蛋白AQP3及紧密连接蛋白ZO-1的表达水平（P<0.05），在保湿类化妆品方面具有较好的开发利用价值。

关键词: 新琼四糖, 保湿, 丝聚蛋白, 水通道蛋白-3, 紧密连接蛋白-1

Abstract: In this research, the final products resulting from the hydrolysis of agar by recombinant agarase AgaP438 were isolated and purified by gel filtration chromatography to investigate the moisturizing activity and potential mechanism of neoagaro-oligosaccharide with single degree of polymerization. The purified products were subjected to analysis, with purity evaluated via ion exchange chromatography (IEC) and the relative molecular mass determined using electrospray time-of-flight mass spectrometry (ESI-TOF-MS). The impact of neoagarotetraose on the viability of keratinocytes was assessed using the CCK-8 assay, while ELISA and immunofluorescence techniques were employed to examine its effects on the expression of filaggrin (FLG), aquaporin-3 (AQP3), and zonula occludens-1 (ZO-1) in keratinocytes. The study findings revealed that the isolation of neoagarotetraose with a relative molecular mass of 630 and a purity of 96.53% was accomplished via Bio-Gel P2 fine gel column, with a yield of 78.4%. Cell experiments demonstrated that neoagarotetraose had no significant impact on the viability of keratinocytes within the mass concentration range of 0.0625－1 mg/mL, and exhibited good biocompatibility. Neoagarotetraose significantly increased the expression of moisturizing-related proteins FLG, AQP3, and ZO-1 in keratinocytes at the mass concentration of 0.125 mg/mL and 0.5 mg/mL (P<0.05). These findings highlighted the development and utilization potential of neoagarotetraose in moisturizing cosmetics, which provided a new direction for the development of cosmetic raw materials.

Key words: neoagarotetraose, moisturizing, filaggrin, aquaporin-3, zonula occludens-1

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