食品科学

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厚壳贻贝多糖的提取工艺优化及体外生物活性研究

钟城城1,曲有乐2,*,陈 荫2   

  1. 1.佳木斯大学食品与药学院,黑龙江 佳木斯 154007;2.浙江海洋学院食品与医药学院,浙江 舟山 316000
  • 出版日期:2014-05-25 发布日期:2014-05-29
  • 通讯作者: 曲有乐
  • 基金资助:

    浙江海洋学院校级专项科研项目(X122X01)

Optimized Extraction and Bioactivity in vitro of Polysaccharides from Mytilus coruscus

ZHONG Cheng-cheng1, QU You-le2,*, CHEN Yin2   

  1. 1. College of Food and Pharmacy, Jiamusi University, Jiamusi 154007, China;
    2. College of Food and Pharmacy, Zhejiang Ocean University, Zhoushan 316000, China
  • Online:2014-05-25 Published:2014-05-29
  • Contact: QU You-le

摘要:

以抗氧化和抗肿瘤活性为指导优化提取工艺,分别采用热水提取,两步联合酶解法(木瓜蛋白酶和胰蛋白酶)从厚壳贻贝中提取和分离纯化具有生物活性的贻贝多糖,对其进行基本理化性质分析和DEAE-Cellulose 52 柱层析分离纯化并进行体外抗氧化、抗肿瘤生物活性研究。通过对热水提取法,单一酶提法和联合酶提取法得到的贻贝多糖粗品进行理化性质分析和活性筛选,发现联合酶解提取得到的贻贝粗多糖在得率、纯度和生物活性上均优于其他方法提取得到的粗多糖,其提取粗多糖的得率为23.69%。联合酶提粗多糖体外抗氧化研究表明其对DPPH自由基、超氧阴离子自由基和脂质过氧化物清除的半最大效应浓度(concentration for 50% of maximal effect,EC50)分别为3.75、5.01 mg/mL和2.41 mg/mL;对体外前列腺癌DU-145细胞48 h,IC50为2.93 mg/mL。通过分离纯化后对各组分的得率和组成分析表明:联合酶提可提高贻贝多糖类糖胺聚糖(MT3)的含量,并达到13.5%,单糖组成主要为Man、GlcN、GlcUA、Gal和Fuc,分子质量约为18 kD。和其他组分相比,MT3 组分表现出良好的体外抗肿瘤活性,对体外前列腺癌DU-145细胞48 h,IC50 为2.71 mg/mL。通过活性追踪和工艺优化结果表明,热水提取后酶解工艺不仅可以降解蛋白提高多糖的纯度,而且还可以显著提高贻贝多糖中类糖胺聚糖的含量,而贻贝类糖胺聚糖是贻贝多糖的主要活性组分,具有良好的抗肿瘤活性。

关键词: 贻贝, 多糖, 提取工艺, 抗氧化, 抗肿瘤

Abstract:

Antioxidant, antitumor polysaccharides were prepared from Mytilus coruscus muscle by hot water extraction
alone or followed by sequential hydrolysis with papain and trypsin, and analyzed for physiochemical properties. After
chromatographic purification on a DEAE-Cellulose 52 column, the antioxidant and antitumor activities in vitro were
investigated. Combined enzymatic hydrolysis, giving rise to a yield of crude polysaccharides of 23.69%, was advantageous
over single enzymatic hydrolysis and hot water extraction as far as the yield, purity and bioactivity of crude polysaccharides
were concerned. The in vitro antioxidant activity of crude polysaccharides obtained from combined enzymatic hydrolysis
showed a concentration for 50% of maximal effect (EC50) of 3.75, 5.01 and 2.41 mg/mL in scavenging DPPH, superoxide
anion and lipid peroxide radicals, respectively, and an IC50 of 2.93 mg/mL against the growth of prostate cancer cells DU-
145 cultured for 48 h. Analysis of the yield and chemical composition of purified polysaccharide fractions indicated that
higher contents of glycosaminoglycan-like polysaccharide (MT3) with a molecular weight of roughly 18 kD in crude
polysaccharides were obtained by combined enzymatic hydrolysis and the monosaccharide composition consisted of Man,
GlcN, GlcUA, Gal and Fuc. Meanwhile, MT3 was more effective against tumor cells DU-145 in vitro with an IC50 of
2.71 mg/mL after 48 h of culture in its presence. The results of activity-guided separation demonstrated that combined
enzymatic hydrolysis after hot water extraction could not only increase the purity of polysaccharides through protein
degradation, but could significantly enhance the content of glycosaminoglycans as well, and glycosaminoglycans were the
main active polysaccharides in Mytilus coruscus muscle that possessed good antitumor activity.

Key words: Mytilus coruscus, polysaccharide, extraction process, antioxidant, antitumor

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