石菖蒲挥发油纤维素酶辅助提取工艺优化及抑制GABA-T活性成分研究
Optimization of cellulase-assisted extraction of volatile oil from Acorus tatarinowii and study on its inhibitory components on GABA-T activity
方成维 1麻秀萍 1陈亮 1李倩1
作者信息
- 1. 贵州中医药大学,贵州贵阳 550025
- 折叠
摘要
目的:打开制约中药中GABA-T配体快速筛选方法的瓶颈.方法:建立中空纤维配体垂钓方法筛选石菖蒲挥发油中γ-氨基丁酸转氨酶(GABA-T)抑制成分.在响应面法优化石菖蒲挥发油酶辅助提取工艺基础上,将GABA-T溶液注射入中空纤维的管腔中,作为"诱饵"垂钓出石菖蒲挥发油中的潜在活性成分,并验证其GABA-T抑制活性.结果:石菖蒲挥发油的最佳提取工艺为纤维素酶添加量3 165 U/g、酶解时间1.8 h、酶解温度45 ℃,此时石菖蒲挥发油收率可达到2.56％,最终鉴定出51个化合物,主要包括烯烃、醇类、醚类、酮类、酚类、酸类等.将3 mg/mL的GABA-T溶液注射入U型中空纤维管的空腔,共孵育160 min,封口后置于石菖蒲挥发油中进行60 kHz超声垂钓20 min,联合LCMS-IT-TOF系统进行分析,成功筛选出的α-细辛醚对GABA-T活性有浓度依赖性的抑制作用,IC5.为57.9 μg/mL.结论:建立的中空纤维配体垂钓方法稳定可行、操作简便.
Abstract
Objective:This study aimed to establish a hollow fiber ligand fishing method for screening γ-Aminobutyrate transaminase(GABA-T)inhibitory component from the volatile oil of Acorus tatarinowii.Methods:Based on optimizing the enzyme assisted extraction process of volatile oil from A.tatarinowii using response surface methodology,GABA-T solution was injected into the hollow fiber lumen and used as a"bait"to fish out the potential active components in A.tatarinowii volatile oil,and its GABA-T inhibitory activity was verified.Results:The results showed that the optimum extraction conditions of volatile oil from A.tatarinowii were as follows:cellulase content 3 165 U/g,enzymolysis time 1.8 h,enzymolysis temperature 45 ℃,and the yield of volatile oil from A.tatarinowii was 2.56％.Finally,51 compounds were identified,mainly including olefins,alcohols,ethers,ketones,phenols,acids,and other compounds.3 mg/mL of GABA-T solution was injected into the cavity of the U-shaped hollow fiber tube,incubated for 160 min,sealed,and placed in the volatile oil of A.tatarinowii for ultrasonic(60 kHz)fishing for 20 min,and then analyzed with LCMS-IT-TOF system.Successfully filtered out α-Asarone has a concentration-dependent inhibitory effect on GABA-T activity,with an IC50 of 57.9 μg/mL.Conclusion:The established hollow fiber ligand fishing method is stable,feasible,and easy to operate.
关键词
石菖蒲/中空纤维/配体垂钓/γ-氨基丁酸转氨酶Key words
Acorus tatarinowii/hollow fiber/ligand fishing/GABA transaminase引用本文复制引用
基金项目
国家自然科学基金(82160803)
贵州中医药大学博士启动基金([2019]58号)
出版年
2024
国家科技期刊平台
NSTL
万方数据