具有酪氨酸酶抑制作用的酵母多肽的分离纯化及分子量分布

为确定具有较好酪氨酸酶抑制作用的酵母多肽的分子量,对酵母多肽粗制品溶液以截留相对分子量为10 KDa、5 KDa、1 KDa的超滤膜进行超滤,采用生化酶法对不同超滤组分进行酪氨酸酶抑制活性检测。结果显示,各多肽组分的活性随相对分子量的减少而增大,活性多肽的相对分子量主要集中在1 KDa以下。对相对分子量1 KDa的多肽组分依次以DEAE-52阴离子交换柱层析法和Sephadex G-15凝胶层柱析法进行分离,最后得到4个洗脱组分,分别富集各洗脱组分并进行酪氨酸酶抑制活性研究。结果表明,组分d-1活性最佳,其IC50值为3.53 mg/mL。经高效液相空间排阻色谱分析,d-1组分多肽的相对分子质量主要集中在200~800 Da。

Purification and molecular weight distribution of tyrosinase inhibitor from yeast peptides

In order to determine the molecular weight distribution of yeast peptides with tyrosinase inhibiton from yeast peptides,the crude products of yeast peptides（ CPYP） solution was successively ultrafiltrated with 10 KDa,5 KDa and 1 KDa ultrafiltration membrane. The tyrosinase inhibition of each fraction was evaluated in vitro. Result showed that the lower molecular-weight peptides fraction shows higher activity of tyrosinase inhibition. The ultrafiltration fraction of high activity（ MW 1 KDa） was further purified using DEAE-52 anion exchange chromatography and Sephadex G-15 gel filtration chromatography. Finally,four fractions were obtained and their tyrosinase inhibition was evaluated respectively. The results indicate that fraction d-1 has higher inhibitory effect on tyrosinase activity with the IC50of 3. 53 mg /mL. The molecular weight distribution of the fraction d-1 is ranged from 200 ～ 800 Da.