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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) chromatography is widely used for large-scale separation of chemical components such as binary sugar mixtures and xylene isomers. In this paper, SMB technology was used to industrialize the isolation of mung bean peptides with high angiotensin-converting enzyme (ACE) inhibitory activity. Using ACE inhibitory activity and yield as an index, the effects of different operating parameters in SMB system on the efficiency of industrialized separation of mung bean peptide were analyzed. The results of 2D-nano LC-MS / MS The amino acid sequence of the active mung bean ACE inhibitory peptide. The results showed that the industrialization of high activity mung bean ACE inhibitory peptides could be achieved by SMB technology. By adjusting the flow rate of various districts to improve product activity and yield. The mung bean peptide ACE isolated under the condition of injection flow rate of 9 m L / min, water flow of 1 mL / min, alcohol flow rate of 11 mL / min and water flow rate of 19 mL / min The inhibition rate was 95.32% and the yield was 86%. Leu-Val-Asn-Pro-Asp-Asp (961.06 u) was obtained by mass spectrometry analysis of isolated high activity mung bean ACE inhibitory peptides with the amino acid sequences of Phe-LeuVal-Asn-Arg- ), Lys-Asp-Asn-Val-Ile-Ser-GluLeu (1 043.13 u).