Peptidomic and Bioinformatic Analysis to Evaluate the Application of Peptides from Latex of Hevea brasiliensis Clone BPM24

Abstract

Peptidomic analysis coupled with mass spectrometry identification was employed to investigate the constitutively expressed peptides in latex serum Phytophthora-resistant rubber tree (Hevea brasiliensis), BPM24. The utilization of proteins and peptides as a strategy to study new methods can lead to a therapeutic application. However, a high-efficiency method to extract peptides from latex serum is not well established. After optimization of latex serum isolation from natural rubber latex, using of lysis solution for peptide extraction exhibited superior serum peptide recovery than using other chemical reagents. Ultrafiltration followed by solid-phase extraction was selected to achieve high extraction yield of low molecular weight proteins and peptides. Mass spectrometric and bioinformatics analysis characterized and predicted the identified protein and peptide contents. All latex serum peptide fractions from C18 reversed-phase chromatography possessed effectiveness of antifungal activity against three tested plant fungal including A. brassicicola, C. lunata and M. grisea with different MIC in the range 250-2,000 μg/ml from different peptide fraction. Moreover, Etotal (E1-E5 pooled together) from lysis solution extraction was found to able to inhibit three tested plant fungal at MIC 500 μg/ml. Moreover, B. cereus and E. faecium were the sensitive gram-positive bacteria with MIC in the range 250-4,000 μg/ml. However, all gram-negative bacteria were not affected by 4,000 μg/ml. Among these tested pathogens, plant fungal was quite sensitive to our peptide product. Etotal from lysis solution extraction will be the best candidate for using further in antifungal testing of other plant fungal both in vitro and in vivo. The further investigation and application will lead to possible use of latex serum as a great biotechnological resource to develop peptide –based product for the treatment of plant infection.