模拟移动床色谱分离纯化绿豆ACE抑制肽

Separation and Purification of ACE-inhibitory Peptides from Mung Bean by Simulated Moving Bed

模拟移动床(Simulated Moving Bed,SMB)色谱多被用于二元糖混合物、二甲苯异构体等化学成分的大批量分离。本文采用SMB技术产业化分离具有较高血管紧张素转化酶(ACE)抑制活性的绿豆肽。以ACE抑制活性和收率为指标,分析SMB系统中不同操作参数对产业化分离绿豆肽效率的影响,并采用二维纳升液质谱联用技术(2D-nano LC-MS/MS)分析高活性绿豆ACE抑制肽的氨基酸序列。结果表明:采用SMB技术可实现高活性绿豆ACE抑制肽的产业化分离。通过调整各区流速提高产品的活性和收率。在进样流速9 m L/min,水洗脱1流速19 m L/min,醇洗脱流速11 m L/min,水洗脱2流速19 m L/min条件下,分离得到的绿豆肽ACE抑制率为95.32%,其收率86%。将分离的高活性绿豆ACE抑制肽进行质谱分析,得出其氨基酸序列分别为Phe-LeuVal-Asn-Arg-Ile(885.1 u)、Phe-Leu-Val-Asn-Pro-Asp-Asp(961.06 u)、Lys-Asp-Asn-Val-Ile-Ser-GluLeu(1 043.13 u)。

Simulated moving bed （SMB） chromatographic separations are known for the efficient separation of well- defined bulk chemicals streams such as binary sugar mixtures or xylene isomers. Therefore, the paper is focusing on the stronger and bulk Angiotensin I-converting enzyme （ACE） inhibitory from Mung bean hydrolysates was purificated by Simulated moving bed （SMB）. Both of the yield and the inhibitory activity of ACE peptide were taken as the indexes, the effects of different operation parameters of SMB system on industrialized separation of mung bean peptides were in- vestigated, and the amino acid sequences of isolated peptides were determined by 2D-nono liquid chromatography mass spectrometry（2D-nonoLC-MS/MS）. High product purities containing the active fraction could be achieved using SMB. Ad- justing the different sector flow rate could increase the bioactive product purity and throughput yield. And the results showed that isolated mung bean peptides possessed higher ACE-inhibitory activity （95.32%） and yield （86%） when op- erating parameters contain feed rate, eluted with water rate, alcohol elution rate and another water elution rate in SMB system were 9 mL-min, 19 mL/min, 11 mL/min, 19 mL/min, respectively. The peptide with the stronger ACE-inhibitory activity had the amino acid sequences of Phe-Leu-Val-Asn-Arg-Ile, Phe-Leu-Val-Asn-Pro-Asp-Asp and Lys-Asp-Asn- Val-Ile-Ser-Glu-Leu, the molecular were 885.1, 961.06 and 1 043.13.