Author:
Wasunan Pimphan,Maneewong Chutamas,Daengprok Wichittra,Thirabunyanon Mongkol
Abstract
Earthworms have long been used as traditional medicine. The purposes of this research were to create bioactive peptides from the unique Amynthas arenulus earthworm (PAAEs) and test their potentials on liver cancer bioprophylactic activity, antioxidant, oxidative stress protection, and immune cell activation. This earthworm had a high protein content ratio, at 55.39%. Besides, PM 35 is one out of 58 bacteria isolated from the earthworm carcasses that exhibited the highest protease and yield protein production which was chosen as the protease-producing bacteria to hydrolyze the protein. The genera were identified by 16S rRNA and 16S–23S rRNA comparison and confirmed as Bacillus velezensis PM 35. The response surface methodology was applied to optimize these hydrolysis parameters, i.e., the enzyme/substrate (E/S) concentration ratio [1%–3% (v/v)] and time (1–3 h) of the hydrolyzing earthworm’s proteins. The optimal hydrolyzing conditions were 3% (v/v) of E/S concentration ratio and 3 h of hydrolysis time, which found protein-hydrolysate yield (24.62%) and degree of hydrolysis (85.45%) as the highest. After being challenged in the gastrointestinal tract-resistant model, these PAAEs (MW <3 and 3–5 kDa) induced liver cancer cell (HepG2) death via apoptotic action modes (cell morphological change and DNA fragmentation). The PAAEs (MW <3 kDa) exhibited significant antioxidant activity via DPPH, ABTS, and FRAP with IC50 values of 0.94, 0.44, and 6.34 mg/ml, respectively. The PAAEs (MW < 3 kDa) were non-cytotoxic and protected the mouse fibroblast cells (L929) against oxidative stress. These PAAEs (MW < 3 kDa, 0.2 mg/ml) stimulated the B lymphocytes (122.3%), and T lymphocytes (126.7%) proliferation. This research suggests that PAAEs can be used in a variety of applications, especially in the food and pharmaceutical industries.
Funder
National Research Council of Thailand
Subject
Microbiology (medical),Microbiology
Reference86 articles.
1. Morphological changes of apoptosis and cytotoxic effects induced by caffeic acid phenethyl ester in AGS human gastric cancer cell line;Amini-Sarteshnizi;J. HerbMed Pharmacol.,2014
2. Characterization of solvent stable extracellular protease from Bacillus koreensis (BK-P21A);Anbu;Int. J. Biol. Macromol.,2013
3. Nutritional and antioxidant evaluation of earthworm powder (Eudrillus euginae);Anitha;Int. Res. J. Pharm.,2012
4. Optimization, purification and characterization of novel thermostable, haloalkaline, solvent stable protease from Bacillus halodurans CAS6 using marine shellfish wastes: a potential additive for detergent and antioxidant synthesis;Annamalai;Bioprocess Biosyst. Eng.,2013
5. In vitro antiproliferative effect of earthworm coelomic fluid of Eudrilus eugeniae, Eisenia foetida, and Perionyx excavatus on squamous cell carcinoma-9 cell line: a pilot study;Augustine;Pharm. Res.,2017
Cited by
5 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献