The fatty acid-binding ability of DUF538 (domain of unknown function 538) protein family in plants was predicted by using molecular docking bioinformatics tools. Two N-terminally derived and engineered peptides (peptide1: KWLVNKIK GKMQ; peptide2: PSICEVGYKDSSVLKFTTKT KVMIWVK) were identified to be the fatty acid-binding elements of DUF538. Using His tag-based recombinant technology, the heterogeneous expression and production of the predicted binding peptides were successfully examined and reported in Escherichia coli cells without affecting the recombinant bacterial cell growth. The recombinant extracts containing peptide1 and peptide2 fusion products exhibited palmitic acid-binding abilities of about 64% and 28%, respectively. As a first report, the presently studied peptides were introduced as novel fatty acid-binding peptides that may be used in medical biotechnology and drug development.