scholarly journals Heterologous expression of a Glyoxalase I gene from sugarcane confers tolerance to several environmental stresses in bacteria

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e5873 ◽  
Author(s):  
Qibin Wu ◽  
Shiwu Gao ◽  
Yong-Bao Pan ◽  
Yachun Su ◽  
Michael P. Grisham ◽  
...  
Keyword(s):  
Abiotic Stresses ◽  
Plasmid Vector ◽  
Stress Conditions ◽  
Glyoxalase I ◽  
Glyoxalase System ◽  
Biotic And Abiotic Stresses ◽  
Expression Plasmid ◽  
Reading Frame ◽  
E Coli ◽  
I Gene

Glyoxalase I belongs to the glyoxalase system that detoxifies methylglyoxal (MG), a cytotoxic by-product produced mainly from triose phosphates. The concentration of MG increases rapidly under stress conditions. In this study, a novel glyoxalase I gene, designated as SoGloI was identified from sugarcane. SoGloI had a size of 1,091 bp with one open reading frame (ORF) of 885 bp encoding a protein of 294 amino acids. SoGloI was predicted as a Ni2+-dependent GLOI protein with two typical glyoxalase domains at positions 28–149 and 159–283, respectively. SoGloI was cloned into an expression plasmid vector, and the Trx-His-S-tag SoGloI protein produced in Escherichia coli was about 51 kDa. The recombinant E. coli cells expressing SoGloI compared to the control grew faster and tolerated higher concentrations of NaCl, CuCl2, CdCl2, or ZnSO4. SoGloI ubiquitously expressed in various sugarcane tissues. The expression was up-regulated under the treatments of NaCl, CuCl2, CdCl2, ZnSO4 and abscisic acid (ABA), or under simulated biotic stress conditions upon exposure to salicylic acid (SA) and methyl jasmonate (MeJA). SoGloI activity steadily increased when sugarcane was subjected to NaCl, CuCl2, CdCl2, or ZnSO4 treatments. Sub-cellular observations indicated that the SoGloI protein was located in both cytosol and nucleus. These results suggest that the SoGloI gene may play an important role in sugarcane’s response to various biotic and abiotic stresses.

Glyoxalase I – structure, function and a critical role in the enzymatic defence against glycation

2003 ◽  
Vol 31 (6) ◽  
pp. 1343-1348 ◽  
Author(s):  
P.J. Thornalley
Keyword(s):  
Metal Ion ◽  
Catalytic Mechanism ◽  
Critical Role ◽  
Glyoxalase I ◽  
Water Molecules ◽  
Glyoxalase System ◽  
E Coli ◽  
Antimalarial Agents

Glyoxalase I is part of the glyoxalase system present in the cytosol of cells. The glyoxalase system catalyses the conversion of reactive, acyclic α-oxoaldehydes into the corresponding α-hydroxyacids. Glyoxalase I catalyses the isomerization of the hemithioacetal, formed spontaneously from α-oxoaldehyde and GSH, to S-2-hydroxyacylglutathione derivatives [RCOCH(OH)-SG→RCH(OH)CO-SG], and in so doing decreases the steady-state concentrations of physiological α-oxoaldehydes and associated glycation reactions. Physiological substrates of glyoxalase I are methylglyoxal, glyoxal and other acyclic α-oxoaldehydes. Human glyoxalase I is a dimeric Zn2+ metalloenzyme of molecular mass 42 kDa. Glyoxalase I from Escherichia coli is a Ni2+ metalloenzyme. The crystal structures of human and E. coli glyoxalase I have been determined to 1.7 and 1.5 Å resolution. The Zn2+ site comprises two structurally equivalent residues from each domain – Gln-33A, Glu-99A, His-126B, Glu-172B and two water molecules. The Ni2+ binding site comprises His-5A, Glu-56A, His-74B, Glu-122B and two water molecules. The catalytic reaction involves base-catalysed shielded-proton transfer from C-1 to C-2 of the hemithioacetal to form an ene-diol intermediate and rapid ketonization to the thioester product. R- and S-enantiomers of the hemithioacetal are bound in the active site, displacing the water molecules in the metal ion primary co-ordination shell. It has been proposed that Glu-172 is the catalytic base for the S-substrate enantiomer and Glu-99 the catalytic base for the R-substrate enantiomer; Glu-172 then reprotonates the ene-diol stereospecifically to form the R-2-hydroxyacylglutathione product. By analogy with the human enzyme, Glu-56 and Glu-122 may be the bases involved in the catalytic mechanism of E. coli glyoxalase I. The suppression of α-oxoaldehyde-mediated glycation by glyoxalase I is particularly important in diabetes and uraemia, where α-oxoaldehyde concentrations are increased. Decreased glyoxalase I activity in situ due to the aging process and oxidative stress results in increased glycation and tissue damage. Inhibition of glyoxalase I pharmacologically with specific inhibitors leads to the accumulation of α-oxoaldehydes to cytotoxic levels; cell-permeable glyoxalase I inhibitors are antitumour and antimalarial agents. Glyoxalase I has a critical role in the prevention of glycation reactions mediated by methylglyoxal, glyoxal and other α-oxoaldehydes in vivo.

Characterization of the glyoxalase I gene from the vascular wilt fungus Verticillium dahliae

2006 ◽  
Vol 52 (9) ◽  
pp. 816-822 ◽  
Author(s):  
A Klimes ◽  
M J Neumann ◽  
S J Grant ◽  
K F Dobinson
Keyword(s):  
Verticillium Dahliae ◽  
Glyoxalase I ◽  
Vascular Wilt ◽  
Glyoxalase System ◽  
Enhanced Sensitivity ◽  
Acid Protein ◽  
Gene Homologue ◽  
I Gene ◽  
Amino Acid Protein

A glyoxalase I gene homologue (VdGLO1) was identified in the vascular wilt fungus Verticillium dahliae by sequence tag analysis of genes expressed during resting structure development. The results of the current study show that the gene encodes a putative 345 amino acid protein with high similarity to glyoxalase I, which produces S-D-lactoylglutathione from the toxic metabolic by-product methylglyoxal (MG). Disruption of the V. dahliae gene by Agrobacterium tumefaciens-mediated transformation resulted in enhanced sensitivity to MG. Mycelial growth of disruption mutants was severely reduced in the presence of 5 mmol/L MG. In contrast, spore production in liquid medium was abolished at 1 mmol/L MG, although not at physiologically relevant concentrations of ≤100 µmol/L. In this first report on the characterization of a glyoxalase I gene in a vascular wilt pathogen, we found that disruption of VdGLO1 had no discernable effect on the pathogenicity of V. dahliae. These data suggest that while the glyoxalase system is necessary for effectively dealing with catastrophic levels of MG, under normal conditions of growth and infection, other MG detoxification pathways in V. dahliae are able to compensate for the absence of the glyoxalase system.Key words: verticillium wilt, glycolytic methylglyoxal pathway, 2-oxoaldehydes.

scholarly journals Different Root Morphological Responses to Phosphorus Supplies in Grafted Pepper

2018 ◽  
Vol 75 (1) ◽  
pp. 59 ◽  
Author(s):  
Leandro PEREIRA-DIAS ◽  
Lidia LOPEZ-SERRANO ◽  
Vicente CASTELL-ZEISING ◽  
Salvador LOPEZ-GALARZA ◽  
Alberto SAN BAUTISTA ◽  
...  
Keyword(s):  
Nutrient Solution ◽  
Root Length ◽  
Abiotic Stresses ◽  
Stress Conditions ◽  
Biotic And Abiotic Stresses ◽  
P Concentration ◽  
Grafting Technique ◽  
Biomass Loss ◽  
Root Response ◽  
Low Input Agriculture

Grafting technique is increasing thanks to its potential to produce plants more efficient and tolerant to biotic and abiotic stresses. Likewise, there is a growing interest in reducing inputs of fertilizers. The development of rootstocks suitable for low input agriculture is conditioned to the understanding of the changes on the root when facing such stresses. Our aim was to evaluate the morphological root response to Phosphorus (P) starvation of a rootstock selected for its good performance under low P conditions. Adige was grafted onto the selected rootstock and grown hydroponically in two different P concentrations, the selft-graft was done as control. Plants were then collected and analysed. Results showed that despite the differences in terms of P concentration among treatment the stress was not enough to cause a great biomass loss. However, there is evidence that individuals showed different root adaptations, modifiying root length, mass and volume, etc, under stress conditions, having the selected rootstock higher root length and volume under low P nutrient solution

scholarly journals Pathogenesis-related protein 1 (PR-1) genes in soybean: genome-wide identification, structural analysis and expression profiling under multiple biotic and abiotic stresses

2021 ◽  
Author(s):  
Fabricio Almeida-Silva ◽  
Thiago M. Venancio
Keyword(s):  
Abiotic Stresses ◽  
Purifying Selection ◽  
Regulatory Elements ◽  
Stress Conditions ◽  
Gene Repertoire ◽  
Biotic And Abiotic Stresses ◽  
Promoter Regions ◽  
Phylogenetic Groups ◽  
Pathogenesis Related Protein ◽  
Pathogenesis Related

Plant pathogenesis-related (PR) proteins are a large group of proteins, classified in 17 families, that are induced by pathological conditions. Here, we characterized the soybean PR-1 (GmPR-1) gene repertoire at the sequence, structural and expression levels. We found 24 GmPR-1 genes, clustered in two phylogenetic groups. GmPR-1 genes are under strong purifying selection, particularly those that emerged by tandem duplications. GmPR-1 promoter regions are abundant in cis-regulatory elements associated with major stress-related transcription factor families, namely WRKY, ERF, HD-Zip, C2H2, NAC, and GATA. We observed that 23 GmPR-1 genes are induced by stress conditions or exclusively expressed upon stress. We explored 1972 transcriptome samples, including 26 stress conditions, revealing that most GmPR-1 genes are differentially expressed in a plethora of biotic and abiotic stresses. Our findings highlight stress-responsive GmPR-1 genes with potential biotechnological applications, such as the development of transgenic lines with increased resistance to biotic and abiotic stresses.

scholarly journals The Molecular Cloning and Expression Analysis of a CYP71 Gene in Ginkgo biloba L.

2016 ◽  
Vol 44 (1) ◽  
pp. 77-84 ◽  
Author(s):  
Xinliang LIU ◽  
Fuliang CAO ◽  
Jinfeng CAI ◽  
Huanli WANG
Keyword(s):  
Molecular Weight ◽  
Expression Analysis ◽  
Ginkgo Biloba ◽  
Abiotic Stresses ◽  
Coli Strain ◽  
Cloning And Expression ◽  
Cytochrome P450 Monooxygenases ◽  
Biotic And Abiotic Stresses ◽  
Reading Frame ◽  
P450 Monooxygenases

Cytochrome P450 monooxygenases (CYPs) are a group of redox proteins that catalyze various oxidative reactions in plant secondary metabolism. To explore the function of the CYP71 gene in Ginkgo biloba under biotic and abiotic stresses, a full-length CYP gene, designated GbCYP71, was first isolated and characterized from leaves of G. biloba. It contained a 1512-bp open reading frame (ORF) encoding 503 amino-acid-deduced polypeptide whose theoretical molecular weight was 56.9 kDa. The genomic DNA sequence of GbCYP71 contained two exons and one intron. The cDNA of GbCYP71 was subcloned in a pET-32a vector and then transformed into E. coli strain BL21 (DE3). A protein with a molecular weight of 76.4 kDa was subsequently identified and found to be consistent with the above theoretical value. Transient expression analysis revealed that the GbCYP71 protein may be located in the G. biloba cell cytoplasm. GbCYP71 was expressed in almost all ginkgo tissues, including leaves, stamens, gynoecia, stems and, preferentially, roots. Expression-profiling analyses revealed that GbCYP71 can be induced by salinity stress and phytohormone signals, including salicylic acid, abscisic acid, methyl jasmonate and ethephon, but is repressed by heat and cold stresses. These results indicate that GbCYP71 mainly functions in responding to biotic and abiotic stresses.

2-Oxoaldehyde metabolism in microorganisms

1989 ◽  
Vol 35 (4) ◽  
pp. 423-431 ◽  
Author(s):  
Kousaku Murata ◽  
Yoshiharu Inoue ◽  
Hae-ik Rhee ◽  
Akira Kimura
Keyword(s):  
Yeast Cells ◽  
Glyoxalase I ◽  
Glyoxalase System ◽  
Metabolizing Enzymes ◽  
Phenotypic Characters ◽  
Metabolism Regulation ◽  
Eukaryotic Microorganisms ◽  
Reductase Gene ◽  
Dividing Cells ◽  
I Gene

The properties of methylglyoxal-metabolizing enzymes in prokaryotic and eukaryotic microorganisms were studied systematically and compared with those of mammalian enzymes. The enzymes constitute a glycolytic bypass and convert methylglyoxal into pyruvate via lactate. The first step in this conversion is catalyzed by glyoxalase I, methylglyoxal reductase, or methylglyoxal dehydrogenase. The regulation of the yeast glyoxalase system was analyzed. The system was closely related to the proliferative states of yeast cells, the activity of the system being high in dividing cells and low in nondividing ones. The gene for the glyoxalase I of Pseudomonas putida and the genes responsible for the activity of glyoxalase I and methylglyoxal reductase in Saccharomyces cerevisiae were cloned and their structural and phenotypic characters studied.Key words: 2-oxoaldehydes metabolism, regulation of glyoxalase system, cloning, glyoxalase I gene, methylglyoxal reductase gene, methylglyoxal metabolism.

Sugar beet M14 glyoxalase I gene can enhance plant tolerance to abiotic stresses

2012 ◽  
Vol 126 (3) ◽  
pp. 415-425 ◽  
Author(s):  
Chuan Wu ◽  
Chunquan Ma ◽  
Yu Pan ◽  
Shilong Gong ◽  
Chenxi Zhao ◽  
...  
Keyword(s):  
Sugar Beet ◽  
Abiotic Stresses ◽  
Glyoxalase I ◽  
Plant Tolerance ◽  
I Gene

scholarly journals Salt-Tolerant Phenomena, Sequencing and Characterization of a Glyoxalase I (Jojo-Gly I) Gene from Jojoba in Comparison with Other Glyoxalase I Genes

Plants ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 1285
Author(s):  
Heba Allah A. Mohasseb ◽  
Mohei El-Din Solliman ◽  
Ibrahim S. Al-Mssallem ◽  
Mohammed M. Ba Abdullah ◽  
Ahmed Saud Alsaqufi ◽  
...  
Keyword(s):  
Amino Acid ◽  
Constitutive Expression ◽  
Pcr Primers ◽  
Glyoxalase I ◽  
Amino Acid Residues ◽  
Salt Tolerant ◽  
Coding Region ◽  
Reading Frame ◽  
Biotic Stresses ◽  
I Gene

Plant response to salt stress and the mechanism of salt tolerance have received major focus by plant biology researchers. Biotic stresses cause extensive losses in agricultural production globally, but abiotic stress causes significant increase in the methylglyoxal (MG) level of GlyoxalaseI (Gly I). Identification of salt-tolerant genes when characterizing their phenotypes will help to identify novel genes using polymerase chain reaction (PCR) to amplify the DNA coding region for glyoxalase I. This method is specific, requiring only genomic DNA and two pairs of PCR primers, and involving two successive PCR reactions. This method was used rapidly and easily identified glyoxalase I sequences as salt-tolerant genes from Jojoba (Simmondsia chinensis (Link) Schneider). In the present study, the glyoxalase I gene was isolated, amplified by PCR using gene-specific primers and sequenced from the jojoba plant, then compared with other glyoxalase I sequences in other plants and glyoxalase I genes like in Brassica napus, ID: KT720495.1; Brassica juncea ID: Y13239.1, Arachis hypogaea; ID: DQ989209.2; and Arabidopsis thaliana L, ID: AAL84986. The structural gene of glyoxalase I, when sequenced and analyzed, revealed that the uninterrupted open reading frame (ORF) of jojoba Gly I (Jojo-Gly I) spans 775 bp, corresponding to 185 amino acid residues, and shares 45.2% amino acid sequence identity to jojoba (Jojo-Gly I). The cloned ORF, in a multicopy constitutive expression plasmid, complemented the Jojo-Gly I, confirming that the encoded Jojo-Gly I in jojoba showed some homology with other known glyoxalase I sequences of plants.

Enhanced Biocatalytic Activity of Recombinant Lipase Immobilized on Gold Nanoparticles

2019 ◽  
Vol 20 (6) ◽  
pp. 497-505 ◽  
Author(s):  
Abeer M. Abd El-Aziz ◽  
Mohamed A. Shaker ◽  
Mona I. Shaaban
Keyword(s):  
Gold Nanoparticles ◽  
Lipolytic Activity ◽  
Market Demand ◽  
Biotechnological Applications ◽  
Lipase Gene ◽  
Expression Plasmid ◽  
E Coli ◽  
Nickel Affinity Chromatography ◽  
Recombinant Production ◽  
Sds Page

Background: Bacterial lipases especially Pseudomonas lipases are extensively used for different biotechnological applications. Objectives: With the better understanding and progressive needs for improving its activity in accordance with the growing market demand, we aimed in this study to improve the recombinant production and biocatalytic activity of lipases via surface conjugation on gold nanoparticles. Methods: The full length coding sequences of lipase gene (lipA), lipase specific foldase gene (lipf) and dual cassette (lipAf) gene were amplified from the genomic DNA of Pseudomonas aeruginosa PA14 and cloned into the bacterial expression vector pRSET-B. Recombinant lipases were expressed in E. coli BL-21 (DE3) pLysS then purified using nickel affinity chromatography and the protein identity was confirmed using SDS-PAGE and Western blot analysis. The purified recombinant lipases were immobilized through surface conjugation with gold nanoparticles and enzymatic activity was colorimetrically quantified. Results: Here, two single expression plasmid systems pRSET-B-lipA and pRSET-B-lipf and one dual cassette expression plasmid system pRSET-B-lipAf were successfully constructed. The lipolytic activities of recombinant lipases LipA, Lipf and LipAf were 4870, 426 and 6740 IUmg-1, respectively. However, upon immobilization of these recombinant lipases on prepared gold nanoparticles (GNPs), the activities were 7417, 822 and 13035 IUmg-1, for LipA-GNPs, Lipf-GNPs and LipAf-GNPs, respectively. The activities after immobilization have been increased 1.52 and 1.93 -fold for LipA and LipAf, respectively. Conclusion: The lipolytic activity of recombinant lipases in the bioconjugate was significantly increased relative to the free recombinant enzyme where immobilization had made the enzyme attain its optimum performance.

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