Keywords

Spirulina Platensis Peptide Anti-Proliferation Activity Enzymatic Hydrolysis Isolation And Purification

Abstract

Using Spirulina as raw material, on the basis of repeated freezing and thawing, homogenization and ultrasound, the protein extraction rates of two different methods, salt extraction and water extraction, were compared. Water-extracted Spirulina protein was used as raw material, using trypsin. , chymotrypsin, pepsin, alkaline protease and papain were subjected to single enzymatic hydrolysis, double enzymatic hydrolysis and triple enzymatic hydrolysis to simulate gastrointestinal digestion in vitro, and 6 enzymatic peptides were obtained. The identification results of the degree of hydrolysis and anti-tumor activity before and after decolorization show that the degree of hydrolysis is not directly related to the anti-tumor activity, and decolorization will reduce the anti-tumor activity of the enzymatic components. Five enzymatic peptides with significant activity were subjected to ultrafiltration, and 9 components were obtained that significantly inhibited the growth of breast cancer and liver cancer cell lines in vitro. The alkaline protease 0~3 K components were Sephadexed. G-15 was separated and purified to obtain component A3, which has a strong inhibitory effect on both MCF-7 and HepG-2. Its IC50 values are 64.59 ¦Ìg/mL and 61.65 ¦Ìg/mL respectively; it has strong selectivity for HepG-2. The IC50 values of the sexually inhibiting components A1, A4 and A5 are 47.67 ¦Ìg/mL, 79.44 ¦Ìg/mL and 69.54 ¦Ìg/mL respectively. The results of this research provide a theoretical basis for the development of medicines and health foods using Spirulina as raw materials. Based on the combination of repeated freeze-thaw cycles, homogenization, and ultrasonic-extraction, the extraction rate between salt-extraction and water-extraction of Spirulina proteins was compared. Taking water-extracted Spirulina proteins as the raw material, six types of Spirulina proteolytic peptides were obtained through enzymolysis of Spirulina proteins using trypsin, chymotrypsin, pepsin, alkaline protease, and papin individually, using alkaline protease and papin together, and using trypsin, pepsin, and chymotrypsin to simulate the digestive processes of the gastrointestinal tract in vitro. Results of the degree of hydrolysis and anti-proliferation effects before and after decolorization suggested that there was no direct correlation between the degree of hydrolysis and the anti-proliferation effects. In addition, decolorization was found to lower the anti-tumor activity of hydrolytic components. . Therefore, five types of proteolytic peptides with good anti-proliferation effects were used for ultrafiltration, providing nine components that could significantly inhibit the growth of both MCF-7 and HepG-2 in vitro. Then, the 0~3 K component of alkaline protease was isolated and purified using a Sephadex G-15 column to obtain the A3 component, which had a strong anti-proliferation effect on MCF-7 and HepG-2 cells, showing IC50 values of 64.59 ¦Ìg/mL and 61.65 ¦Ìg/mL, respectively. Additionally, another three components, A1, A4, and A5, were obtained, which had strong selective anti-proliferation effects on HepG-2 cells, with IC50 values of 47.67 ¦Ìg/mL, 79.44 ¦Ìg/mL, and 69.54 ¦Ìg/ mL, respectively. These results provide a theoretical basis for the development of new drugs and health foods using Spirulina platensis as raw materials.

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Original research done by Wang Zhujun

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