Reduction of quinones and phenoxy radicals by extracellular glucose dehydrogenase from Glomerella cingulata suggests a role in plant pathogenicity

Sygmund, Christoph; Klausberger, Miriam; Felice, Alfons K. G.; Ludwig, Roland

doi:10.1099/mic.0.051904-0

Cited by 46 publications

(61 citation statements)

References 33 publications

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“…For GDH expressed by G. cingulata (produced according to [10]) the temperature optimum was 48°C. In a more detailed investigation using the ThermoFAD technique to derive thermal unfolding transition values (Tm) for different pH values and buffer substances (Table 2), recGc GDH showed a pH-dependent thermal stability with the highest Tm values in the acidic range of 4.5 to 6.4.…”

Section: Resultsmentioning

confidence: 99%

“…Two big producers of glucose biosensors, Abbott and Bayer, already implemented FAD-dependent GDHs in some of their products. A novel member of the small family of FAD-dependent GDHs was recently discovered in the plant pathogenic fungus Glomerella cingulata (anamorph Colletotrichum gloeosporoides ) and characterized [10]. It is an extracellular, glycosylated enzyme showing a narrow substrate specificity with β-D-glucose and D-xylose as substrates, which are oxidized at the anomeric carbon atom.…”

Section: Introductionmentioning

confidence: 99%

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Heterologous overexpression of Glomerella cingulata FAD-dependent glucose dehydrogenase in Escherichia coli and Pichia pastoris

et al. 2011

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BackgroundFAD dependent glucose dehydrogenase (GDH) currently raises enormous interest in the field of glucose biosensors. Due to its superior properties such as high turnover rate, substrate specificity and oxygen independence, GDH makes its way into glucose biosensing. The recently discovered GDH from the ascomycete Glomerella cingulata is a novel candidate for such an electrochemical application, but also of interest to study the plant-pathogen interaction of a family of wide-spread, crop destroying fungi. Heterologous expression is a necessity to facilitate the production of GDH for biotechnological applications and to study its physiological role in the outbreak of anthracnose caused by Glomerella (anamorph Colletotrichum) spp.ResultsHeterologous expression of active G. cingulata GDH has been achieved in both Escherichia coli and Pichia pastoris, however, the expressed volumetric activity was about 4800-fold higher in P. pastoris. Expression in E. coli resulted mainly in the formation of inclusion bodies and only after co-expression with molecular chaperones enzymatic activity was detected. The fed-batch cultivation of a P. pastoris transformant resulted in an expression of 48,000 U L-1 of GDH activity (57 mg L-1). Recombinant GDH was purified by a two-step purification procedure with a yield of 71%. Comparative characterization of molecular and catalytic properties shows identical features for the GDH expressed in P. pastoris and the wild-type enzyme from its natural fungal source.ConclusionsThe heterologous expression of active GDH was greatly favoured in the eukaryotic host. The efficient expression in P. pastoris facilitates the production of genetically engineered GDH variants for electrochemical-, physiological- and structural studies.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Heterologous overexpression of Glomerella cingulata FAD-dependent glucose dehydrogenase in Escherichia coli and Pichia pastoris

et al. 2011

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“…The role of peroxide generated by plant pathogens is diverse [44]. For example, it may interfere with plant defense compounds by direct reduction, as was recently proposed for a FAD-dependent glucose dehydrogenase from the Pezizomycotina Glomerella cingulata that causes anthracnose on various fruits [45]. …”

Section: Discussionmentioning

confidence: 99%

Compositions of fungal secretomes indicate a greater impact of phylogenetic history than lifestyle adaptation

et al. 2014

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BackgroundSince the first fungal genome sequences became available, investigators have been employing comparative genomics to understand how fungi have evolved to occupy diverse ecological niches. The secretome, i.e. the entirety of all proteins secreted by an organism, is of particular importance, as by these proteins fungi acquire nutrients and communicate with their surroundings.ResultsIt is generally assumed that fungi with similar nutritional lifestyles have similar secretome compositions. In this study, we test this hypothesis by annotating and comparing the soluble secretomes, defined as the sets of proteins containing classical signal peptides but lacking transmembrane domains of fungi representing a broad diversity of nutritional lifestyles. Secretome size correlates with phylogeny and to a lesser extent with lifestyle. Plant pathogens and saprophytes have larger secretomes than animal pathogens. Small secreted cysteine-rich proteins (SSCPs), which may comprise many effectors important for the interaction of plant pathogens with their hosts, are defined here to have a mature length of ≤ 300 aa residues, at least four cysteines, and a total cysteine content of ≥5%. SSCPs are found enriched in the secretomes of the Pezizomycotina and Basidiomycota in comparison to Saccharomycotina. Relative SSCP content is noticeably higher in plant pathogens than in animal pathogens, while saprophytes were in between and closer to plant pathogens. Expansions and contractions of gene families and in the number of occurrences of functional domains are largely lineage specific, e.g. contraction of glycoside hydrolases in Saccharomycotina, and are only weakly correlated with lifestyle. However, within a given lifestyle a few general trends exist, such as the expansion of secreted family M14 metallopeptidases and chitin-binding proteins in plant pathogenic Pezizomycotina.ConclusionsWhile the secretomes of fungi with similar lifestyles share certain characteristics, the expansion and contraction of gene families is largely lineage specific, and not shared among all fungi of a given lifestyle.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-15-722) contains supplementary material, which is available to authorized users.

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“…Most FAD-GDH enzymes are extracted from fungi [16][17][18], and there are few reports on heterologous expression in E. coli, which usually results in no expression or formation of inclusion bodies [14]. The highest reported productivity of FAD-GDH was 13,000 U/L and a specific activity of 52 U/mg with 40 mM glucose as substrate [14].…”

Section: Introductionmentioning

confidence: 99%

Expression, characterization and mutagenesis of an FAD-dependent glucose dehydrogenase from Aspergillus terreus

Yang

Huang

Wang

et al. 2015

Enzyme and Microbial Technology

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Reduction of quinones and phenoxy radicals by extracellular glucose dehydrogenase from Glomerella cingulata suggests a role in plant pathogenicity

Cited by 46 publications

References 33 publications

Heterologous overexpression of Glomerella cingulata FAD-dependent glucose dehydrogenase in Escherichia coli and Pichia pastoris

Heterologous overexpression of Glomerella cingulata FAD-dependent glucose dehydrogenase in Escherichia coli and Pichia pastoris

Compositions of fungal secretomes indicate a greater impact of phylogenetic history than lifestyle adaptation

Expression, characterization and mutagenesis of an FAD-dependent glucose dehydrogenase from Aspergillus terreus

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