Resistance to antibiotics is a serious worldwide crisis that possess the challenge of development new chemical tools to tackle it. Random-sequence peptide mixtures (natural Antimicrobial peptides analogues) are composed of a hydrophobic and cationic amino acids in binary ratio with controlled chain length and stereochemistry. Although the sequences are random, high and efficient microbial growth inhibition achieved in low concentrations. The influence of amino acids identity on the antimicrobial activity against varied bacteria has explored. The random sequenced peptides mixtures have a rapid mode of action against the bacteria including entry into the bacterial cells. The promising potential of use in medical and food aspects has studied by exposure pathogens or food associated bacteria to the random sequenced peptide mixtures. These compounds have great potential as new antimicrobial agents for several applications.