METHODS: ACE inhibitory proteins were isolated from P. cystidiosus based on the bioassay guided purification steps, i.e. ammonium sulphate precipitation, reverse phase high performance liquid chromatography and size exclusion chromatography. Active fraction was then analysed by LC-MS/MS and potential ACE inhibitory peptides identified were chemically synthesized. Effect of in vitro gastrointestinal digestions on the ACE inhibitory activity of the peptides and their inhibition patterns were evaluated.
RESULTS: Two potential ACE inhibitory peptides, AHEPVK and GPSMR were identified from P. cystidiosus with molecular masses of 679.53 and 546.36 Da, respectively. Both peptides exhibited potentially high ACE inhibitory activity with IC50 values of 62.8 and 277.5 μM, respectively. SEC chromatograms and BIOPEP analysis of these peptides revealed that the peptide sequence of the hexapeptide, AHEPVK, was stable throughout gastrointestinal digestion. The pentapeptide, GPSMR, was hydrolysed after digestion and it was predicted to release a dipeptide ACE inhibitor, GP, from its precursor. The Lineweaver-Burk plot of AHEPVK showed that this potent and stable ACE inhibitor has a competitive inhibitory effect against ACE.
CONCLUSION: The present study indicated that the peptides from P. cystidiosus could be potential ACE inhibitors. Although these peptides had lower ACE inhibitory activity compared to commercial antihypertensive drugs, they are derived from mushroom which could be easily obtained and should have no side effects. Further in vivo studies can be carried out to reveal the clear mechanism of ACE inhibition by these peptides.
METHODS: Essential oils obtained by steam distillation were analyzed by gas chromatography-mass spectrometry (GC-MS). The antimicrobial activity of the essential oils was evaluated against four bacteria: Bacillus cereus (B. cereus), Staphylococcus aureus (S. aureus), Escherichia coli (E. coli), and Pseudomonas aeruginosa (P. aeruginosa); and two fungi: Candida albicans (C. albicans) and Cyptococcus neoformans (C. neoformans), using disc-diffusion and broth microdilution methods.
RESULTS: Cycloisolongifolene, 8,9-dehydro formyl (35.29%) and dihydrocostunolide (22.51%) were the major compounds in C. aeruginosa oil; whereas caryophyllene oxide (18.71%) and caryophyllene (12.69%) were the major compounds in C. mangga oil; and 2,6,9,9-tetramethyl-2,6,10-cycloundecatrien-1-one (60.77%) and α-caryophyllene (23.92%) were abundant in Z. cassumunar oil. The essential oils displayed varying degrees of antimicrobial activity against all tested microorganisms. C. mangga oil had the highest and most broad-spectrum activity by inhibiting all microorganisms tested, with C. neoformans being the most sensitive microorganism by having the lowest minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values of 0.1 μL/mL. C. aeruginosa oil showed mild antimicrobial activity, whereas Z. cassumunar had very low or weak activity against the tested microorganisms.
CONCLUSIONS: The preliminary results suggest promising antimicrobial properties of C. mangga and C. aeruginosa, which may be useful for food preservation, pharmaceutical treatment and natural therapies.