食品科学 ›› 2012, Vol. 33 ›› Issue (1): 136-140.doi: 10.7506/spkx1002-6630-201201027

• 生物工程 • 上一篇    下一篇

藻蓝蛋白酶解肽的分离纯化及其细胞毒活性

王雪青,邓 伟,杨进芳,毛羽聪,史中明   

  1. 天津市食品与生物技术重点实验室,天津商业大学生物技术与食品科学学院
  • 收稿日期:2011-07-21 修回日期:2012-01-05 出版日期:2012-01-15 发布日期:2012-01-12
  • 通讯作者: 王雪青 E-mail:wxqing@tjcu.edu.cn
  • 基金资助:

    天津市基础研究重点项目(08JCZDJC16600);天津商业大学SRT 项目(2011-57)

Purification and Cytotoxicity of C-Phycocyanin(C-PC) from Spirulina platensis and Its Tryptic Peptides

WANG Xue-qing,DENG Wei,YANG Jin-fang,MAO Yu-cong,SHI Zhong-ming   

  1. Tianjin Key Laboratory of Food Biotechnology, College of Biotechnology and Food Science,
    Tianjin University of Commerce, Tianjin 300134, China
  • Received:2011-07-21 Revised:2012-01-05 Online:2012-01-15 Published:2012-01-12
  • Contact: Xue-qing WANG E-mail:wxqing@tjcu.edu.cn

摘要:

研究钝顶螺旋藻(Spirulina platensis)藻蓝蛋白(C-phycocyanin,C-PC)及其胰蛋白酶水解肽的分离纯化。采用反复冻融和超声破碎法破碎细胞,用28~55g/100mL硫酸铵沉淀反复盐析获得纯度(A620nm/A280nm)为2.19的藻蓝蛋白,再通过羟基磷灰石(HA)柱层析和Sephacryl S-200 HR凝胶层析对其进行纯化,得到纯度(A620nm/A280nm)为3.89藻蓝蛋白。纯化后的藻蓝蛋白在40℃条件下经胰蛋白酶酶解60min后,用DEAE-Sepharose Fast Flow柱层析对酶解肽产物进行分离,收集得到4组藻蓝蛋白酶解肽。采用MTT方法,研究藻蓝蛋白、酶解混合液、分离的4个酶解肽组分对肿瘤细胞系HeLa和293T增殖的影响。结果显示:肽组分1和4对HeLa细胞的抑制率分别为37.71%和47.04%,而混合肽和藻蓝蛋白抑制率分别为34.02%和26.03%,因此活性肽组分1和4显示出较好的肿瘤抑制效果,而这两种活性肽组分对正常细胞293T并无细胞毒活性,特别是组分4效果最明显,是极具开发潜力的保健产品。

关键词: 藻蓝蛋白, 纯化, 酶解肽, 细胞毒活性

Abstract:

This paper deals with the separation, purification and cytotoxicity of C-phycocyanin (C-PC) from Spirulina platensis and its tryptic peptides. Repeated freezing and thawing coupled with ultrasonic treatment was used for disrupting the cell wall of Spirulina platensis. The purity (A620nm/A280nm) of C-PC was 2.19 after fractional precipitation by 28-55 g/100 mL (NH4)2SO4 and could reach 3.89 after further purification by sequential chromatography on hydroxylapatite (HA) column and Sephacryl S-200 HR gel column. The purified C-PC was hydrolyzed by trypsin at 40 ℃ for 60 min. Four peptides were obtained from the hydrolysate of C-PC by DEAE-Sepharose Fast Flow column chromatography. The cytotoxicity of C-PC and its hydrolysate as well as the 4 peptides on HeLa and 293T cells was evaluated by MTT assay. The results showed that the inhibition rates of peptide fractions I and IV, C-PC hydrolysate and C-PC on the growth of HeLa cells were 37.71%, 47.04%, 34.02% and 26.03%, respectively. Therefore, peptide fractions I and IV revealed obvious suppressive effect on the proliferation of cancer cells, while neither of them had cytotoxicity on 293T cells. Moreover, peptide fraction IV had the strongest tumor suppression activity, indicating a great potential to be developed as health-care products.

Key words: C-phycocyanin, purification, hydrolyzed peptide, cytotoxicity

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