Fermented bean foods are a crucial source of fibrinolytic enzymes. The presented study aimed to purify, characterize, and chemically modify Bacillus velezensis SN-14 fibrinolytic enzyme. The fibrinolytic enzyme was purified using CTAB/isooctane/hexyl alcohol/n-butyl alcohol reverse micellar system, and the purified enzyme was chemically modified to improve its enzymatic activity and stability. Enzyme activity recovery and the purification fold for this enzyme were 44.5 ± 1.9% and 4.93 ± 0.05 fold, respectively. SDS-PAGE results showed that the molecular weight of the purified fibrinolytic enzyme was around 28 kDa. Besides, the optimum temperature and pH of the purified fibrinolytic enzyme were 37 °C and 8-9, respectively. Fe2+, mPEG5000, and pepsin were used for chemical modification and for improving the activity and stability of the purified enzyme. Thermal and acid-base stability of chemically modified enzymes increased significantly, whereas enzymatic activity increased by 7.3 times. After 30 d of frozen storage, the modified enzyme's activity was remarkably lower (33.2%) than the unmodified enzyme (60.6%). The current study on B. velezensis SN-14 fibrinolytic enzyme and chemical modification method using Fe2+, mPEG5000, and pepsin provide a reference for developing fibrinolytic drugs and foods.
Keywords: Acid-base stability; Bacillus velezensis SN-14; Chemical modification; Fibrinolytic enzyme; Frozen storage stability; Purification; Thermal stability.
Copyright © 2021 Elsevier B.V. All rights reserved.