| Lysobacter can produce a variety of antagonistic substances for microorganism, which are broadly active against bacteria, fungi, single-celled algae and nematodes, so it is an important and new type of gram-negative bacteria groups with biocontrol and medicinal value. As a new class of natural active microbial resource, Lysobacter Enzymogenes C3 as a model strain of Lysobacter, whose products have broad spectrum antifungal activities. Currently, Lysobacter Enzymogenes C3 is just culturing with shaking flasks, for application needs to optimize the medium formulation, and with industrialized cheap recipe to achieve large-scale industrial product. Because of yellow product in the Lysobacter Enzymogenes C3 fermentation process, when it applied to bamboo, clothing mildew, the pigment attached affects its appearance and value, hence need to develop high efficient decoloring technology to produce colorless fermentation bacteria. Using activated carbon or resin adsorbent decolorize Lysobacter Enzymogenes C3 to produce the highly active colorless C3 microbial inoculum is environmentally safe, low cost, simple and the raw materials sources is widely, etc. It can be applied to paper, bamboo, flowers, clothing and fruit and vegetables to avoid mould, and keep the proper color and quality of the article, has significantly meaning.In this paper, Lysobacter enzymogenes C3 is the research object. First, using cheap industrial medium formulation design process optimization to ferment Lysobacter Enzymogenes C3, promote their efficient to synthesis active products. Then optimize the operation conditions of activated carbon adsorbent, diatomite, resins and others, to maintain a high activity and colorless inoculants. Then apply the efficient fungicide to avoid paper mold, fresh cut flowers lily liquid, and rot ptoofing of pine mildew corrosion and other process.1 Research on industrialization with cheap medium to compound a large number of synthetic Lysobacter Enzymogenes C3 active products. Employ single factor optimization, plackett-Burman (P-B) design and response surface analysis to optimization design method to obtain the optimum fermentation medium:corn flour 1.18%, soybean meal 1.10%, NaCl 0.92%, CaCl20.04%, Na2MoO40.01%. Compared to former recipe the optimization formulations significantly enhanced antibacterial activity, the activity of inhibition Bacillus subtilis increased 16.7%, the activities of resistance to Magnaporthe grisea increased by 26.5%.2 Develop high activity and colorless C3 microbial agent. Reasearch on the production process of natural decoloring antimold agent. Using regression analysis and experimental verification, the best conditions for C3 fermentation medium is to add 1.6% granular activated carbon, the bleaching temperature is 27℃, the adsorption time is 1.2h, and the decolorization pH is 8.0.3 High activity and colorless C3 fermentation agents used in paper mildew. Application of C3 agent for paper mouldproof research trials, purified the 5 kinds of mildew fungus from the mouldy paper, with the identification of its morphology and molecular biology, they were Z-1 Alternaria alternata, Z-2 Aspergillus aculeatus, Z-3 Stachybotrys echinata, Z-4Aspergillus niger, Z-5 Stachybotrys chartarum. Determination the activity of cellulase enzyme and laccase of the isolated mildew fungus, the results show it has significantly activity on paper degradation. The C3 agents have significant antifungal activity on the isolated mildew strains, further measured of the capacity to avoid paper mold it shows good effect. Compared with the control group, the treatment group of C3 agents has good mechanical properties, while tensile strength of paper was significantly decreased in the control group, the paper was serious corrupted. So, C3 antifungal agents has significant effect on paper.4 C3 agents applied to fresh cut flowers lily. Through the preservation effect study of liquid formulations for different fresh cut flowers lily, compared to tap water (control group 1),C3 agents treatment formula can extend the life of cut flowers for 8 days, fresh weight changes in rate of 8.68%; water balance increased 2.63;chlorophyll a, b content increased by 37.2%,13.0%; protein content increased 12.3%; T-SOD activity was increased 33.3%; hydrogen peroxide enzyme (CAT) activity increased 23.6%; malondialdehyde (MDA) content decreased 30.1%. Compared to Ⅷ number (control 2),C3 agents treatment formula can extend the life of cut flowers for 7.5days, fresh weight changes in rate of 7.98%; water balance increased 2.86; chlorophyll a, b content increased by 8.9%、30.0%; protein content increased 10.4%; T-SOD activity was increased 31.7%; hydrogen peroxide enzyme (CAT) activity increased 16.4%; malondialdehyde (MDA) content decreased 29.7%. The results illustrated C3 agents can slow down lily’s cell aging, improve the freshness of lily cut flowers, extend viewing period, and promote their growth.5 C3 antifungal preservative agents applied to pine. Isolated and purified 13 kinds of mildew fungus from mildew pine wood, with the morphological and molecular biological identification, they were:S1 Penicillium citrinum、S2 Phomopsis bougainvilleicola、S3 Penicillium vanoranjei、S4 Trametes corrugata、S5 Nectria haematococca、S6 Gibberella moniliformis、S7 Paecilomyces parvisporus、S8 Fusarium proliferatum、S9 Rhizopus oryzae、S10 Aspergillus fumigate、S11 Trichoderma harzianum、S12 Trichoderma atroviride、S13 Trichoderma asperellum. Measure the cellulase activity and laccase activity of the isolated mildew fungus, all molds were showing a corresponding activity, and have a certain ability to degrade pine. C3 agents also have a strong antibacterial activity to pine mold, inhibited the growth of mildew fungus isolated from pine. In indoor pine mold proof experiment, the pine mildew proof effect of C3 agent was good. Compared to the treatment group and control group pine surface under the scanning electronic microscope, the surface of the control group covered with pine mold spores and hyphae, while the pine surface treated with C3 agents was clean and sterile, showing C3 has obvious antifungal effect for pine. Determinated the timber weight loss, the mass loss rate was 12.3% in the control group, the experimental group (C3 treatment) mass loss rate was 6.8%. Determinated the chemical composition of wood (extractables), the hot water timber, content of cellulose dissolution, cold water extract,1% sodium hydroxide solution and extract were increased in the control group than C3 agents group, and the benzene extraction and an alcohol content of lignin dissolution surge has great increase. The control group showed severe decay pine structure more vulnerable to collapse, leachables surge. C3 agents treatment can increase the mould proof capacity of pine, wooden structure to maintain normal, less dissolution of the various components. The agents can significantly improve the ability of pine mildew rot.In this paper, through the research of the development of Lysobacter enzymogenes C3 agent of industrialization of cheap medium formula, and develop the decoloring technology to produce colorless microbial inoculum, and C3 antifungal agents is successfully used in paper, flowers, bamboo, wood mold. It can lay the foundation for the preparation of C3 antifungal agents and antifungal applications. |
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