Cloning, Expression, Purification and Initial Analysis of a Novel Pectate Lyase <i><scp>P</scp>cpel1</i> from <i><scp>P</scp>hytophthora capsici</i>

Fu, Li; Wang, Hui Z.; Feng, Bao Zhen; Zhang, Xiu G.

doi:10.1111/jph.12059

Cited by 4 publications

(3 citation statements)

References 56 publications

(99 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Alignment of P. parasitica full-length PL1 proteins showed that they have a number of conserved residues, with pairwise identity ranging from 21% to 100%. Two PL1 proteins from P. capsici, Pcpel1 and Pcpel2 [NCBI: FJ213434, FJ213435] have been identified as pectate lyases, having activity against unesterified polygalacturonic acid (PGA) [ 120 , 121 ]. Pcpel2 has 74-90% amino acid identity with three P. parasitica PL1s [PPTG_12901, 12902 and 20388] while Pcpel1 has 87% identity with one P. parasitica PL1 [PPTG_18908], suggesting that these P. parasitica proteins will act on unesterified HG.…”

Section: Resultsmentioning

confidence: 99%

Bioinformatic characterisation of genes encoding cell wall degrading enzymes in the Phytophthora parasitica genome

2014

View full text Add to dashboard Cite

BackgroundA critical aspect of plant infection by the majority of pathogens is penetration of the plant cell wall. This process requires the production and secretion of a broad spectrum of pathogen enzymes that target and degrade the many complex polysaccharides in the plant cell wall. As a necessary framework for a study of the expression of cell wall degrading enzymes (CWDEs) produced by the broad host range phytopathogen, Phytophthora parasitica, we have conducted an in-depth bioinformatics analysis of the entire complement of genes encoding CWDEs in this pathogen’s genome.ResultsOur bioinformatic analysis indicates that 431 (2%) of the 20,825 predicted proteins encoded by the P. parasitica genome, are carbohydrate-active enzymes (CAZymes) involved in the degradation of cell wall polysaccharides. Of the 431 proteins, 337 contain classical N-terminal secretion signals and 67 are predicted to be targeted to the non-classical secretion pathway. Identification of CAZyme catalytic activity based on primary protein sequence is difficult, nevertheless, detailed comparisons with previously characterized enzymes has allowed us to determine likely enzyme activities and targeted substrates for many of the P. parasitica CWDEs. Some proteins (12%) contain more than one CAZyme module but, in most cases, multiple modules are from the same CAZyme family. Only 12 P. parasitica CWDEs contain both catalytically-active (glycosyl hydrolase) and non-catalytic (carbohydrate binding) modules, a situation that contrasts with that in fungal phytopathogens. Other striking differences between the complements of CWDEs in P. parasitica and fungal phytopathogens are seen in the CAZyme families that target cellulose, pectins or β-1,3-glucans (e.g. callose). About 25% of P. parasitica CAZymes are solely directed towards pectin degradation, with the majority coming from pectin lyase or carbohydrate esterase families. Fungal phytopathogens typically contain less than half the numbers of these CAZymes. The P. parasitica genome, like that of other Oomycetes, is rich in CAZymes that target β-1,3-glucans.ConclusionsThis detailed analysis of the full complement of P. parasitica cell wall degrading enzymes provides a framework for an in-depth study of patterns of expression of these pathogen genes during plant infection and the induction or repression of expression by selected substrates.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-15-785) contains supplementary material, which is available to authorized users.

show abstract

Section: Resultsmentioning

confidence: 99%

Bioinformatic characterisation of genes encoding cell wall degrading enzymes in the Phytophthora parasitica genome

2014

View full text Add to dashboard Cite

show abstract

“…They cleave the glycosidic bonds of two saccharide units via a β-elimination mechanism ( Collmer and Keen, 1986 ; Herron et al, 2000 ). Studies have shown that pectate lyases contribute to virulence of bacteria, fungi, nematodes, and oomycetes ( Wegener, 2002 ; Fu et al, 2013 ; Cho et al, 2015 ; Chen et al, 2021 ). A study of pectate lyase genes PL1 , PL15 , PL16 , and PL20 from P. capsici showed that overexpression of these genes in a mildly virulent strain transformed it to a highly aggressive strain ( Fu et al, 2015 ).…”

Section: Discussionmentioning

confidence: 99%

Genome analysis of Phytophthora cactorum strains associated with crown- and leather-rot in strawberry

et al. 2023

View full text Add to dashboard Cite

Phytophthora cactorum has two distinct pathotypes that cause crown rot and leather rot in strawberry (Fragaria × ananassa). Strains of the crown rot pathotype can infect both the rhizome (crown) and fruit tissues, while strains of the leather rot pathotype can only infect the fruits of strawberry. The genome of a highly virulent crown rot strain, a low virulent crown rot strain, and three leather rot strains were sequenced using PacBio high fidelity (HiFi) long read sequencing. The reads were de novo assembled to 66.4–67.6 megabases genomes in 178–204 contigs, with N50 values ranging from 892 to 1,036 kilobases. The total number of predicted complete genes in the five P. cactorum genomes ranged from 17,286 to 17,398. Orthology analysis identified a core secretome of 8,238 genes. Comparative genomic analysis revealed differences in the composition of potential virulence effectors, such as putative RxLR and Crinklers, between the crown rot and the leather rot pathotypes. Insertions, deletions, and amino acid substitutions were detected in genes encoding putative elicitors such as beta elicitin and cellulose-binding domain proteins from the leather rot strains compared to the highly virulent crown rot strain, suggesting a potential mechanism for the crown rot strain to escape host recognition during compatible interaction with strawberry. The results presented here highlight several effectors that may facilitate the tissue-specific colonization of P. cactorum in strawberry.

show abstract

“…Pel A gene from Aspergillus nidulans has been successfully expressed in Bacillus subtilis [99] and E. coli [100]. Pectate lyase gene designated as Pcpel2 and Pcpel1from Phytophthora capsicin have been cloned and expressed in E. coli and Pichia pastoris respectively [101,102] revealing its significant role in pathogenesis. The open reading frame of Pcpel1I gene is 1233 bp and encodes 410 amino acid polypeptide, including 21 residues long amino terminal signal sequence.…”

Section: Fungal Pectate Lyasesmentioning

confidence: 99%

Molecular Biology of Microbial Pectate Lyase: A Review

Dubey

Yadav

Kumar

et al. 2016

BBJ

View full text Add to dashboard Cite

Pectate lyase represents an important member of pectinase group of enzymes responsible for the pathogenesis and softening of plant tissues. It also has role in fruit juice clarification and in retting of natural fibers. The biochemical characterization of pectate lyases from diverse microbial sources and plants along with an insight to the protein structure has been dealt earlier but there is a lack of exclusive review on the molecular biology of pectate lyases. This review tries to fill the gap by highlighting the various aspects of molecular biology of microbial pectate lyases especially the cloning and expression of pectate lyase genes from diverse sources attempted so far. The topics covered in this review are a brief description about enzymes associated with degradation of pectin, its classification, applications, updated information about the biochemical characterization of microbial pectate lyases and cloning and expression of microbial pectate lyase genes.

show abstract

Cloning, Expression, Purification and Initial Analysis of a Novel Pectate Lyase Pcpel1 from Phytophthora capsici

Cited by 4 publications

References 56 publications

Bioinformatic characterisation of genes encoding cell wall degrading enzymes in the Phytophthora parasitica genome

Bioinformatic characterisation of genes encoding cell wall degrading enzymes in the Phytophthora parasitica genome

Genome analysis of Phytophthora cactorum strains associated with crown- and leather-rot in strawberry

Molecular Biology of Microbial Pectate Lyase: A Review

Contact Info

Product

Resources

About