International Journal of Biological Macromolecules p. 740 - 751 (2019)
Update date:2022-08-11
Topics:
Qian, JunQing
Zhao, ChangYan
Tong, Jun
Jiang, ShengLan
Zhang
Lu, ShiYong
Guo, Hui
Frontal affinity chromatography (FAC) combined with enzyme has been widely used for drug screening. In this paper, the effect of target enzyme activity on screening of bioactive compounds was studied through applying FAC. Trypsin with different degree of inactivation were prepared as target enzyme by thermal denaturation. Their primary structure was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and use Fourier transform infrared (FTIR) and ultraviolet-visible (UV–vis) spectroscopy to detect group structure. Ultimately, it was found that the main structure of enzyme with decreased activity remained unchanged. The oxymatrine and matrine which can interact with trypsin were selected to study their binding to trypsin with different activities in FAC. The results showed that oxymatrine and matrine had a significant difference in the breakthrough volume among seven kinds of columns prepared by trypsins with different activities, at the different concentration. It indicated that trypsins with different activities in FAC could combine with oxymatrine and matrine. The binding constant (Kd) variation between oxymatrine, matrine and trypsin with different activities are 5.520 ± 0.038 and 3.577 ± 0.071, within error range, which indicated that the activity of target enzyme with primary unchanged structure has no effect on screening of bioactive components by FAC.
View MoreContact:86 21 3772 9386
Address:Rm.1803,Starry Bldg.1,1505 Meijiabang Road,Shanghai 201620 China
NanJing Rate Biochemicals CO., LTD
Contact:+86-25-84931986
Address:NO. 1 Hongjing Road,Jiangning Science Park,Nanjing,China
Chongqing Changfeng Chemical Co., Ltd.
website:http://www.changfengchem.com
Contact:+86-23-67896333
Address:30th Floor, Longhu Ziduxingzuo Building A, 1st Branch,YuSong Rd., Yubei District, Chongqing, China
Changzhou Kingyo Chemical Corporation Ltd.
website:http://www.kingyochem.com
Contact:+86-519-85105717
Address:19# Wuqing North Road, Changzhou , Jiangsu, China
Contact:+0086-21-61984905-1
Address:Building 60,Zimian Park, LongYang industrial Area, 1515Nong,Yuandong Road Fengxian District, Shanghai ,China
Doi:10.1007/s11746-013-2220-5
(2013)Doi:10.1021/ja00483a050
(1978)Doi:10.1016/j.tetlet.2020.152061
(2020)Doi:10.1002/zaac.201600259
(2016)Doi:10.1016/S0006-2952(97)00379-1
(1998)Doi:10.1002/asia.201700950
(2017)