In silico directed evolution of Anabas testudineus AtMP1 antimicrobial peptide to improve in vitro anticancer activity

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Various studies have demonstrated that directed evolution is a powerful tool in
enhancing protein properties. In this study, directed evolution was used to enhance
the efficacy of synthesised Anabas testudineus AtMP1 antimicrobial peptides (AMPs)
in inhibiting the proliferation of cancer cells. The modification of antimicrobial
peptides (AMPs) and prediction of peptide properties using bioinformatic tools were
carried out using four databases, including ADP3, CAMP-R3, AMPfun, and ANTICP.
One modified antimicrobial peptide (AMP), ATMP6 (THPPTTTTTTTTTTTTTAAPARTT),
was chosen based on its projected potent anticancer effect, taking into
account factors such as amino acid length, net charge, anticancer activity score, and
hydrophobicity. The selected AMPs were subjected to study in deep-learning databases,
namely ToxIBTL and ToxinPred2, to predict their toxicity. Furthermore, the allergic
properties of these antimicrobial peptides (AMPs) were verified by utilising AllerTOP
and AllergenFP. Based on the results obtained from the database study, it was projected
that antimicrobial peptides (AMPs) demonstrate a lack of toxicity towards human
cells that is indicative of the broader population. After 48 hours of incubation, the
IC50 values of ATMP6 against the HS27 and MDA-MB-231 cell lines were found
to be 48.03 0.013 mg/ml and 7.52 0.027 mg/ml, respectively. The IC50 values
of the original peptide ATMP1 against the MDA-MB-231 and HS27 cell lines were
determined to be 59.6 0.14 mg/ml and 8.25 0.14 mg/ml, respectively, when
compared. Furthermore, the results indicated that the injection of ATMP6 induced
apoptosis in the MDA-MB-231 cell lines. The present investigation has revealed new
opportunities for advancing novel targeted peptide therapeutics to tackle cancer.