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Study of the Lipopeptide Antibiotics Iturins from Bacillus Thuringiensis

Author: GuoLiQing
Tutor: GuanXiong
School: Fujian Agriculture and Forestry University
Course: Biochemistry and Molecular Biology
Keywords: Bacillus thuringiensis PCR Lipopeptide antibiotic Antibacterial HPLC
CLC: S476.1
Type: Master's thesis
Year: 2010
Downloads: 197
Quote: 3
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Abstract


Particularly various Bacillus subtilis (Bacillus subtilis) during the growth and metabolism can produce different types of antibacterial substances, including non-ribosomal synthesis of lipopeptide antibiotics (lipopeptide antibiotics) Biological surfactin (surfactin), Fongyuan Su (fengycin) and Iturin (iturin) and so on. Which iturin family antibiotics in agriculture, biological, medical, etc. to be more widely applied. This study attempts to use PCR, fungal inhibition assay and HPLC are three ways to identify production iturin strains, view from Bacillus subtilis kinship similar but has not yet found producing iturin family of antibiotics Bacillus thuringiensis found in production iturinA so has antibacterial lipopeptide antibiotic strains. Use the GenBank logged ituD primers were designed to save our laboratory Bacillus thuringiensis, Bacillus subtilis, Bacillus amyloliquefaciens and Bacillus cereus strains such as total DNA as a template for PCR amplification was obtained 9 PCR-positive strains. On the cloned gene was sequenced and ituD Blast sequence analysis showed that the cloned gene in GenBank logged iturin lipopeptide antibiotic gene family has more than 95% homology. These nine strains fermentation broth after acid precipitation, methanol extraction and other steps to obtain lipopeptides crude extracts and suppress banana anthracnose (Colletotrichum musae), Fusarium oxysporum (Fusarium oxysporum), Botrytis gray mold ( Botrytis cinerea) and rice sheath blight (Rhizoctonia solani) and other common plant pathogenic fungi determination. The results showed that the extracted lipopeptide crude extract, Bacillus subtilis EPL8, Bacillus amyloliquefaciens TB2 on the four kinds of fungal mycelia growth have a strong inhibitory effect on Bacillus subtilis BS1 fungus Colletotrichum musae and Botrytis cinere mycelial growth inhibition, and Bacillus thuringiensis BRC-XQ16 the right Colletotrichum musae inhibited mycelial growth. Inhibit fungal hyphae will be part of this four bacterial lipopeptides crude extract was purified through reversed-phase HPLC analysis showed that, EPL8 and TB2 lipopeptide extracts, the highest peak of the three components and iturinA standard samples are close to the corresponding peaks inferred EPL8 and TB2 in lipopeptide extract might exist iturinA structurally similar components. BS1 lipopeptide extract component peak retention time is almost the highest standard samples with iturinA highest peak in the same retention time consistent inferred that this component is probably the highest concentrations iturinA standard sample components. BRC-XQ16 in the two components of the highest peak retention time iturinA standard sample in the first two components of similar retention time, the same speculation, BRC-XQ16 of these two kinds of component structure is also similar iturinA. According to the experimental speculated that this experiment EPL8, TB2, BS1 and BRC-XQ16 of four strains can produce iturinA or other similar structure with iturinA iturin family members. The experimental results for these strains produced lipopeptide antibiotics further structural determination of the foundation, but also to broaden the application of Bacillus thuringiensis foundation.

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CLC: > Agricultural Sciences > Plant Protection > A variety of control methods > Biological control > The use of microbial pathogens
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