The producer of the anti-mycotic principle was identified as Enterococcus faecalis based on its physiological and biochemical characteristic. Based on the 16S rDNA gene sequence, the strain was identified as E. faecium[19]. Further, using the primers EM1A and EM1B [20], an amplicon of approximately 685 base pairs was

observed on 1.2% (w/v) agarose gel confirming the strain to be E. faecium. However, this strain reduced potassium PD0332991 solubility dmso tellurite and produced black colour colonies, indicating the species E. faecalis. The two wild type isolates (DI and WI) of the pathogenic indicator organism were identified as C. albicans based on 18S ribotyping. The sequences of the DI and WI isolates showed closest homology (99%) to the sequences of C. albicans M60302.YSASRSUA and AJ005123, respectively. Determination of inhibitory spectrum The susceptibilities of various multidrug resistant C. albicans strains to growth inhibition by the supernatant as well as dialysed concentrate of E. faecalis are presented in Table 1. The supernatant and dialysed concentrate also showed inhibitory activity against one wild type C. albicans strain (DI) isolated from a diabetic patient from Goa. Amongst these strains, maximum activity was observed against C. albicans strains MTCC 183, MTCC 3958, MTCC 7315, and NCIM 3471 and minimum

activity was observed against wild type C. albicans (DI) (Figure 1a, b, c) and C.krusei (data not shown). The biological activity of ACP at different dilutions is shown in Figure 1 (d and e) against MTCC 183. Table Mitomycin C 1 Inhibitory spectrum of anti- Candida protein ACP against different indicator organisms Strain Identified

organisms Indicator organisms Zone of inhibition 210 E. faecalis Yersinia intermedia (AGM 108–5) 25 mm     Candida albicans >18 mm (NCIM 3471, MTCC183, MTCC 7315, MTCC 227 and MTCC 3958)   Dialysed Concentrate MTCC183 and MTCC 7315 55 mm, 47 mm     Wild type C. albicans (DI) Teicoplanin 13 mm Figure 1 a. Biological activity of ACP against C. albicans (MTCC 7315). b. Biological activity of ACP against C. albicans (MTCC 183) after 85% ammonium sulfate fractionation, The zone of inhibition was detected in 85% palette dissolved in 20 mmol sodium phosphate buffer pH 8.0, but activity was not detected in supernatant. c. Mild biological activity of ACP against wild type C. albicans (DI) isolated from a diabetic patient in BITS Goa. d and e. Different concentration of dialyzed concentrate of ACP showing zone of inhibition against a lawn of C. albicans MTCC 183. Antimicrobial activity of cell wall and cytoplasmic extracts The antimicrobial activity of the cell wall and cytoplasmic extracts of E. faecalis was determined using a cut-well agar assay on MGYP and BHI plates. No zone of inhibition was produced against C.