Partial Purification and Biochemical Evaluation of Protease Fraction (MA-1) from <i>Mycoleptodonoides aitchisonii</i> and Its Fibrinolytic Effect

• Main Authors: Sung-Ho Lee (Author), Seung-Yub Song (Author), Jun-Hui Choi (Author), Seung Kim (Author), Hyo-Jeong Lee (Author), Jin Woo Park (Author), Dae-Hun Park (Author), Chun-Sik Bae (Author), Seung-Sik Cho (Author)
• Format: Book
• Published: MDPI AG, 2023-08-01T00:00:00Z.
• Subjects: article
• Online Access: Connect to this object online.

Summary

The antioxidative proteolytic fraction, MA-1, was partially purified from <i>Mycoleptodonoides aitchisonii</i>. MA-1 was purified to homogeneity using a two-step procedure, which resulted in an 89-fold increase in specific activity and 42.5% recovery. SDS-PAGE revealed two proteins with a molecular weight of 48 kDa. The zymography results revealed proteolytic activity based on the MA-1 band. MA-1 was found to be stable in the presence of Na⁺, Ca²⁺, Fe³⁺, K⁺, and Mg²⁺. MA-1 was also stable in methanol, ethanol, and acetone, and its enzyme activity increased by 15% in SDS. MA-1 was inhibited by ethylenediaminetetra-acetic acid or ethylene glycol tetraacetic acid and exerted the highest specificity for the substrate, MeO-Suc-Arg-Pro-Tyr-pNA, for chymotrypsin. Accordingly, MA-1 belongs to the family of chymotrypsin-like metalloproteins. The optimum temperature was 40 °C and stability was stable in the range of 20 to 35 °C. The optimum pH and stability were pH 5.5 and pH 4-11. MA-1 exhibited stronger fibrinolytic activity than plasmin. MA-1 hydrolyzed the Aα, Bβ, and γ chains of fibrinogen within 2 h. MA-1 exhibited an antithrombotic effect in animal models. MA-1 was devoid of hemorrhagic activity at a dose of 80,000 U/kg. Overall, our results show that <i>M. aitchisonii</i> produces an acid-tolerant and antioxidative chymotrypsin-like fibrinolytic enzyme, and <i>M. aitchisonii</i> containing MA-1 could be a beneficial functional material for the prevention of cardiovascular diseases and possible complications.

Item Description

10.3390/antiox12081558
2076-3921