Purification and characterization of an exopolygalacturonase produced by Fusarium oxysporum f. sp. radicis lycopersici

VAZQUEZ, C

doi:10.1016/0378-1097(93)90465-e

Cited by 3 publications

(6 citation statements)

References 19 publications

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“…The purification of the EXOPG protein of the isolate r6 of FORL revealed one band of 66 kDa by SDS‐PAGE. Deglycosylation reduced its molecular weight to 50 kDa, in agreement with previously reported data (Vázquez et al. 1993).…”

Section: Resultssupporting

confidence: 93%

“…Fungi were stored as microconidial suspensions in 15% glycerol at −80°C. In vitro methods to analyse pgx2 and pgx1 expression by Northern blot were basically the same as those reported in Vázquez et al. (1993).…”

Section: Organisms and Culture Conditionsmentioning

confidence: 99%

“…1997b). However, the most abundant PG enzyme of the FORL r6 isolate produced in pectin cultures was purified and showed an exo mode of action (Vázquez et al. 1993).…”

Section: Introductionmentioning

confidence: 99%

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Characterization and in vitro expression patterns of an exopolygalacturonase encoding gene from Fusarium oxysporum f.sp. radicis lycopersici

et al. 2003

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Aims: In this work, we report the isolation, characterization and expression pattern in in vitro cultures of an EXOPG encoding gene (pgx2), a novel EXOPG encoding gene of Fusarium oxysporum f.sp. radicis lycopersici, responsible for foot crown and root rot disease in tomato plants. The gene was compared with other fungal polygalacturonases (PGs) previously reported. Methods and Results: Partial sequences of the purified EXOPG native protein were used to design primers that amplified a genomic fragment by PCR. The amplified genomic fragment was used as a probe to screen a genomic library. One isolated clone was analysed. The complete genomic, cDNA and the deduced amino acid sequences were compared with other fungal EXOPGs and ENDOPGs. Regulation of pgx2 expression was analysed by Northern blot in in vitro cultures supplemented with different carbon sources. Conclusions: Pgx2 was present as single copy in the haploid genome of several Fusarium species. PGX2 showed the conserved amino acid motifs typical of PGs and those reported for fungal EXOPGs. Pgx2 was regulated at transcriptional level showing similar expression pattern to other EXOPG encoding gene (pgx1) when the fungus was cultured on different carbon sources suggesting a coordinate expression of both genes. This similarity would be supported by the presence of common putative regulatory motifs in the upstream regions of both genes. Significance and Impact of the Study: This study reports the analysis of a novel EXOPG gene of the tomato pathogen F. oxysporum f.sp. radicis lycopersici, a contribution to the understanding of the role of cell‐wall‐degrading enzymes produced by fungi during pathogenesis.

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Section: Resultssupporting

confidence: 93%

Section: Organisms and Culture Conditionsmentioning

confidence: 99%

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Characterization and in vitro expression patterns of an exopolygalacturonase encoding gene from Fusarium oxysporum f.sp. radicis lycopersici

et al. 2003

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“…• C (Foda et al 1984;Vazquez et al 1993;Martin et al 2004). The purified PG from R. oryzae MTCC 1987 does not loose its activity appreciably up to 30…”

Section: )mentioning

confidence: 99%

Purification and characterization of an exo-polygalacturonase secreted by Rhizopus oryzae MTCC 1987 and its role in retting of Crotalaria juncea fibre

et al. 2012

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An acidic polygalacturonase (PG) secreted by Rhizopus oryzae MTCC-1987 in submerged fermentation condition has been purified to electrophoretic homogeneity using ammonium sulphate fractionation and anion exchange chromatography on diethylaminoethyl cellulose. The purified enzyme gave a single protein band in sodium dodecyl sulphatepolyacrylamide gel electrophoresis analysis with a molecular mass corresponding to 75.5 kDa. The Km and kcat values of the PG were 2.7 mg/mL and 2.23 × 10 3 s −1 , respectively, using citrus polygalacturonic acid as the substrate. The optimum pH of the purified PG was 5.0 and it does not loose activity appreciably if left for 24 hours in the pH range from 5.0 to 12.0. The optimum temperature of purified enzyme was 50• C and the enzyme does not loose activity below 30• C if exposed for two hours. The purified enzyme showed complete inhibition with 1 mM Ag + , Hg 2+ and KMnO4, while it was stimulated to some extent by Co 2+ . The purified PG exhibited retting of Crotalaria juncea fibre in absence of ethylenediaminetetraacetic acid.

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“…ciceri [6] and Sclerotinia sclerotiorum exoPG [14], and the M, (74000) was comparable to that of the exoPGs from F. oxysporum f.sp. radicis-lycopersici [8], F oxysporum fsp. ciceri [6], A. rabiei [7], S. sclerotiorum [14], and Colletotrichum lindemuthianum [4].…”

Section: Biochemical Characterization Of Pg2mentioning

confidence: 99%

Purification and characterization of an exo-polygalacturonase from the tomato vascular wilt pathogen Fusarium oxysporum f.sp. lycopersici

Pietro

1996

FEMS Microbiology Letters

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An exo-polygalacturonase (EC 3.2.1.15) was purified to apparent homogeneity from cultures of Fusarium oxysporum fsp.lycopersici on synthetic medium supplemented with citrus pectin, using preparative isoelectric focusing. The enzyme, denominated PG2, had an apparent A4, of 74 000 Da upon SDS-PAGE. The pI of the main PG2 isoform was 4.5, and pH and temperature optima were 5.0 and WC, respectively. PG2 hydrolyzed polygalacturonic acid in an exo-manner, as demonstrated by anaysis of degradation products. The enzyme was N-glycosylated. The N-terminal amino acid sequence, L-A-F-N-V-P-S-K-P-P, has no identity to other known polygalacturonases.

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Purification and characterization of an exopolygalacturonase produced by Fusarium oxysporum f. sp. radicis lycopersici

Cited by 3 publications

References 19 publications

Characterization and in vitro expression patterns of an exopolygalacturonase encoding gene from Fusarium oxysporum f.sp. radicis lycopersici

Characterization and in vitro expression patterns of an exopolygalacturonase encoding gene from Fusarium oxysporum f.sp. radicis lycopersici

Purification and characterization of an exo-polygalacturonase secreted by Rhizopus oryzae MTCC 1987 and its role in retting of Crotalaria juncea fibre

Purification and characterization of an exo-polygalacturonase from the tomato vascular wilt pathogen Fusarium oxysporum f.sp. lycopersici

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