Antimicrobial Activity Synergism Between Bacillus Lytic Enzyme L27 And Beta-1, 3-1, 4-glucanase

With the broad application of the antibiotics, resistant pathogens are increasingly public health concern. Seeking novel antibiotics becomes imperative. Some enzymes with lytic action can degraded cell wall and show antibacterial and antifungal activity against pathogens. As the characteristic structure and compose of the microbial wall, the complex enzymes can kill microorganism more effectively. The complex enzymes can have broader-spectrum and more effective antibacterial.To develop more potent protease, a clone was isolated and classified to Bacillus sp. The fermentation products of this strain were separated and identified as Lytic enzyme L27 and beta-1,3-1,4-glucanase(EC 3.2.1.6). Bioassay showed that the purified sample had remarkable inhibitory effect on a broad spectrum of microorganisms, such as Staphylococcus aureaus, Escherichia coli and Schizosaccharomyces pombe. The two proteins were cloned and expressed in E.coli. The recombinant proteins were purified via Ni-sepharose affinity chromatography, and the inhibitory effect of the two recombinant proteins and their compound on Candida albicans, Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Staphylococcus aureaus and Micrococcus luteus were detected. The bioassay result showed that the compound of the two recombinant proteins have stronger inhibitory effect than either.Our result provides directed evidence for the synergism between the two proteins: the lytic activity of glucanase needs the present of the alkaline protease, and vice versa. The structure of the two molecules might provide a lead scaffold to design and screen smaller and more potent druggable compound to combat the increasingly drug resistant pathogen.