Chitinolytic Enterobacter agglomerans Antagonistic to Fungal Plant Pathogens

Chernin, Leonid; Ismailov, Z. F.; Haran, Shoshan; Chet, I.

doi:10.1128/aem.61.5.1720-1726.1995

Cited by 317 publications

(131 citation statements)

References 35 publications

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“…For neither protein was a clear N-terminal signal peptide predicted, suggesting they are cytoplasmic proteins. Overexpressed and purified HexI showed activity towards all three tested chitin analogs (Table 2), which is consistent with previous reports on N-acetylglucosaminidases (Chernin et al, 1995). We were not able to StrepTag-purify HexII from overexpressing E. coli cells (Table 2).…”

Section: Resultssupporting

confidence: 89%

Identification and characterization of genes underlying chitinolysis in Collimonas fungivorans Ter331

Fritsche

Boer

Gerards

et al. 2008

FEMS Microbiology Ecology

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Through a combinatorial approach of plasposon mutagenesis, genome mining, and heterologous expression, we identified genes contributing to the chitinolytic phenotype of bacterium Collimonas fungivorans Ter331. One of five mutants with abolished ability to hydrolyze colloidal chitin carried its plasposon in the chiI gene coding for an extracellular endochitinase. Two mutants were affected in the promoter of chiP-II coding for an outer-membrane transporter of chitooligosaccharides. The remaining two mutations were linked to chitobiose/N-acetylglucosamine uptake. Thus, our model for the Collimonas chitinolytic system assumes a positive feedback regulation of chitinase activity by chitin degradation products. A second chitinase gene, chiII, coded for an exochitinase that preferentially released chitobiose from chitin analogs. Genes hexI and hexII showed coding resemblance to N-acetylglucosaminidases, and the activity of purified HexI protein towards chitin analogs suggested its role in converting chitobiose to N-acetylglucosamine. The hexI gene clustered with chiI, chiII, and chiP-II in one locus, while chitobiose/N-acetylglucosamine uptake genes colocalized in another. Both loci contained genes for conversion of N-acetylglucosamine to fructose-6-phosphate, confirming that C. fungivorans Ter331 features a complete chitin pathway. No link could be established between chitinolysis and antifungal activity of C. fungivorans Ter331, suggesting that the bacterium's reported antagonism towards fungi relies on other mechanisms.

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Section: Resultssupporting

confidence: 89%

Identification and characterization of genes underlying chitinolysis in Collimonas fungivorans Ter331

Fritsche

Boer

Gerards

et al. 2008

FEMS Microbiology Ecology

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“…These results indicated that expression of Chi92 on the cell surface enhanced the ability for inhibition of fungal growth in vitro. It was reported that the chitinase-producing bacteria, including Aeromonas caviae (Inbar & Chet, 1991), Enterobacter agglumerans (Chernin et al, 1995), and Serratia marcescens (Ordentlich et al, 1988), are potential biocontrol agents against phytopathogenic fungi. The chiA gene from S. marcescens has been expressed in E. coli, and the chitinase was shown to be effective in the reduction of disease incidence caused by S. rolfsii and R. solani (Shapira et al, 1989).…”

Section: Antifungal Activitiesmentioning

confidence: 99%

Cell surface display of Chi92 onEscherichia coliusing ice nucleation protein for improved catalytic and antifungal activity

Wu¹,

Tsai

Chen³

2006

FEMS Microbiology Letters

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The gene encoding chitinase 92 (Chi92) from Aeromonas hydrophila JP10 has been displayed on the cell surface of Escherichia coli using the N-terminal region of ice nucleation proteins (INPN) as an anchoring motif. Immunofluorescence microscopy confirmed that Chi92 was anchored on the cell surface. Western blot analysis further identified the synthesis of INP derivatives containing the N-terminal domain INPN-Chi92 fusion protein of the expected size (112 kDa). Whole cell enzyme assay indicated that the displayed Chi92 showed enhanced catalytic activity toward colloidal chitin. In addition, the Chi92-displayed cells exhibited inhibitory effects on the mycelial growth of phytopathogenic fungi, including Fusarium decemcellulare, Sclerotium rolfsii, Rhizoctonia solani kuhn, and Fusarium oxysporum f.sp. melonis. This study suggested that the INP-based display systems can be used to express a large protein (90 kDa Chi92) on the cell surface of E. coli without growth inhibition. In addition, the display of chitinase on the cell surface may provide an attractive method for the development of biocontrol agents against phytopathogenic fungi.

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“…In controlling fungal plant pathogens, a variety of mechanisms contribute to the biocontrol activity of microbes. Cell-wall-degrading enzymes (CWDEs), such as b-1,3-glucanases, cellulases, proteases, and chitinases, are involved in the antagonistic activity of some biological control agents against phytopathogenic fungi [12][13][14][15][16]. In particular, numerous correlations between fungal antagonism and bacterial production of chitinases and/or b-1,3-glucanases have been noted [17].…”

Section: Introductionmentioning

confidence: 99%

Production of an antifungal protein for control of Colletotrichum lagenarium by Bacillus amyloliquefaciens MET0908

Kim¹

2004

FEMS Microbiology Letters

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A plant pathogenic fungus, Colletotrichum lagenarium, causing watermelon anthracnose, was isolated from naturally infected leaves, stems, and fruits of watermelon. A bacterial strain, MET0908, showing a potent antifungal activity against C. lagenarium, was isolated from soil. An antifungal protein was purified by 30% ammonium sulfate saturation and concentrated using Centricon 10, DEAE-Sepharose(TM) Fast Flow column and Sephacryl S-100 gel filtration chromatography. The molecular weight of the purified protein was estimated as 40 kDa by SDS-PAGE. The purified protein was stable at 80 degrees C for 20 min and exhibited a broad spectrum of antifungal activity against various plant pathogenic fungi. Confocal microscopy image analysis and scanning electron microscopy showed that the protein acted on the cell wall of C. lagenarium. The purified antifungal protein exhibited beta-1,3-glucanase activity. The N-terminal amino acid sequence of the purified protein was determined as Ser-Lys-Ile-x-Ile-Asn-Ile-Asn-Ile-x-Gln-Ala-Pro-Ala-Pro-x-Ala. A search of the sequence with NCBI BLAST showed no significant homology with any known proteins, suggesting that the purified protein may be novel.

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Chitinolytic Enterobacter agglomerans Antagonistic to Fungal Plant Pathogens

Cited by 317 publications

References 35 publications

Identification and characterization of genes underlying chitinolysis in Collimonas fungivorans Ter331

Identification and characterization of genes underlying chitinolysis in Collimonas fungivorans Ter331

Cell surface display of Chi92 onEscherichia coliusing ice nucleation protein for improved catalytic and antifungal activity

Production of an antifungal protein for control of Colletotrichum lagenarium by Bacillus amyloliquefaciens MET0908

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