Title:Optimization of the Enzymolysis Conditions for Scorpion Peptides and Evaluation of its Antitumor Activity
VOLUME: 11 ISSUE: 1
Author(s):Fangwen Jiao, Jihui Wang, Yuhong Liu, Yan Zou, Wei Jia, Panpan Zhang, Jing Sun, Youwei Xu and Junxiang Wang
Affiliation:School of Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan 250355, China.
Keywords:Orthogonal design experiment, enzymolysis conditions, anti-tumor activity, cell apoptosis, cell cycle analysis.
Abstract:Trypsin, chymotrypsin and elastase were combined to hydrolyze scorpion
Buthus marstesii Karsch powders to prepare scorpion peptides (SP). Orthogonal
design experiment L16 (44) was used to optimize variables of enzymolysis
temperature, time, pH and enzyme concentration. Degree of hydrolysis (DH) and
inhibition rate (IR) of A549 cells were selected as analysis indicators. The optimum
enzymolysis conditions were determined as follows: enzymolysis temperature 40°C,
time 3h, pH 8.5 and enzyme concentration of each 2000U. The anti-tumor activity of
SP was determined on a panel of representative cell lines (A549, MCF7 and EC109) using MTT assay. In vitro,
SP significantly inhibited the proliferation of A549, which was more obvious than MCF-7 and EC109. Further
experiments on cell apoptosis and cell cycle analysis revealed that SP induce apoptosis and arrest cell cycle progression in
S phase.