Antimicrobial activity of a protein purified from the latex of Hevea brasiliensis on oral microorganisms

Abstract

This study aimed at screen for antimicrobial activity present in the non-rubber constituents of rubber latex of Hevea brasiliensis against various microbial strains. An antimicrobial protein, hevein was extracted from the bottom fraction after centrifugation and purified by acetone fractionation and anion exchange chromatography on a DEAE-Sepharose Fast Flow column. This procedure was more efficient and rapid than the previously described procedures. The antimicrobial activity was investigated and revealed that hevein, a small (4.7 kDa) cysteine-rich protein, had strong antimicrobial activity, especially against Candida spp. including Candida albicans, Candida tropicalis and Candida krusei. The MIC80 value for hevein was as low as 12 microg ml(-1) with C. tropicalis ATCC 750. Higher MIC80 values were obtained for C. albicans ATCC 10231 (95 microg ml(-1)) and C. krusei ATCC 6258 (190 microg ml(-1)). To confirm the antifungal activity, hevein also inhibited the growth of those fungi in a disk diffusion assay and its inhibition was enhanced when a Hevea latex protease inhibitor was also included. Hevein at a concentration of 30 microg ml(-1) also caused a Ca2+-dependent aggregation of C. tropicalis yeast cells. These data indicate that hevein can inhibit the growth of certain potential oral fungal pathogens.

Semi-quantitative RT-PCR analysis of transcripts encoding protease inhibitor inHevea brasiliensisMuell. Arg latex

“…In a matter of fact, twenty-five per cent of the proteins in C-and B-serums are known as antimicrobial agents in Hevea brasiliensis. Most of these proteins have the activity of chitinase/lysozyme which can degrade fungal and bacterial cell wall components such as chitin and peptidoglycan, respectively [8,11]. However, among these known proteins, small size proteins known as protease inhibitor were less studied in Hevea latex.…”

“…Limited numbers of research have reported the isolation and characterization of protease inhibitor from the latex of two Hevea tree clones RRIM 600 and PB 260 [11][12][13][14][15]. In molecular levels, high numbers of Expressed Sequence Tags (ESTs) encoding protease inhibitors were predicted in laticifers [10].…”

Valorisation of Proteins from Rubber Tree

“…According to our current knowledge, the protein with the most prospective application is hevein for antimicrobial or antifungal agents [7,70]. The other protein with potential application is the 43 kDa patatin-homologue [10,12], due to its similarity with patatin.…”

Purification, characterization, and bioassay of putative protease inhibitors from Hevea brasiliensis latex

“…Isolation and characterization of protease inhibitors from various plant species have been carried out using affinity column chromatography (Bijina et al, 2011;Popovic et al, 2013;Al-Assadi, 2013, Bunyatang et al, 2013Chaudhary et al, 2008;Sabotič et al, 2012). In H. brasiliensis, several types of protease inhibitors have also been successfully extracted from latex (Bangrak & Chotigeat, 2011;Kanokwiroon et al, 2008;Sritanyarat et al, 2006). Bunyatang et al (2013) has demonstrated the inhibitory activity of Hevea protease inhibitors against the mycelium growth of Phytophthora palmivora.…”

“…In matter of fact, twentyfive percent of the proteins in C-and B-serums are known as antimicrobial agents in H. brasiliensis. Most of these proteins have the activity of chitinase/lysozyme which can degrade fungal and bacterial cell wall components such as chitin and peptidoglycan, respectively (Martin, 1991;Kanokwiroon et al, 2008). Other lutoid-contained proteins such as chitinase, glucanase, hevein, hevamine have antifungal activity against Botrytis cinerea, Fusarium culmorum, Fusarium oxysporum, Phycomyces blakesleeanus, and Trichoderma hamatum (Bokma et al, 2000;Churngchow et al, 1995, Chye & Cheung, 1995, Parijs et al, 1991.…”