Purification and characterization of biologically active 1,4-linked α-d-oligogalacturonides after partial digestion of polygalacturonic acid with endopolygalacturonase

Spiro, Mark D.; Kates, Keith; Koller, Alan L.; O’Neill, Malcolm A.; Albersheim, Peter; Darvill, Alan G.

doi:10.1016/0008-6215(93)84237-z

Cited by 73 publications

(42 citation statements)

References 16 publications

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“…The solution was incubated with agitation for 16 h at 4°C and then centrifuged at 7,500 ϫ g for 15 min. The supernatant was diluted, and the absorbance at 235 nm was measured (28). The absorbance of the fermentation supernatant preparation of E. coli strain KO11 at 72 h was used as the baseline.…”

Section: Methodsmentioning

confidence: 99%

Addition of Genes for Cellobiase and Pectinolytic Activity in Escherichia coli for Fuel Ethanol Production from Pectin-Rich Lignocellulosic Biomass

Edwards

Henriksen

Yomano

et al. 2011

Appl Environ Microbiol

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Ethanologenic Escherichia coli strain KO11 was sequentially engineered to contain the Klebsiella oxytoca cellobiose phosphotransferase genes (casAB) as well as a pectate lyase (pelE) from Erwinia chrysanthemi, yielding strains LY40A (casAB) and JP07 (casAB pelE), respectively. To obtain an effective secretion of PelE, the Sec-dependent pathway out genes from E. chrysanthemi were provided on a cosmid to strain JP07 to construct strain JP07C. Finally, oligogalacturonide lyase (ogl) from E. chrysanthemi was added to produce strain JP08C. E. coli strains LY40A, JP07, JP07C, and JP08C possessed significant cellobiase activity in cell lysates, while only strains JP07C and JP08C demonstrated extracellular pectate lyase activity. Fermentations conducted by using a mixture of pure sugars representative of the composition of sugar beet pulp (SBP) showed that strains LY40A, JP07, JP07C, and JP08C were able to ferment cellobiose, resulting in increased ethanol production from 15 to 45% in comparison to that of KO11. Fermentations with SBP at very low fungal enzyme loads during saccharification revealed significantly higher levels of ethanol production for LY40A, JP07C, and JP08C than for KO11. JP07C ethanol yields were not considerably higher than those of LY40A; however, oligogalacturonide polymerization studies showed an increased breakdown of biomass to small-chain (degree of polymerization, <6) oligogalacturonides. JP08C achieved a further breakdown of polygalacturonate to monomeric sugars, resulting in a 164% increase in ethanol yields compared to those of KO11. The addition of commercial pectin methylesterase (PME) further increased JP08C ethanol production compared to that of LY40A by demethylating the pectin for enzymatic attack by pectin-degrading enzymes.

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Section: Methodsmentioning

confidence: 99%

Addition of Genes for Cellobiase and Pectinolytic Activity in Escherichia coli for Fuel Ethanol Production from Pectin-Rich Lignocellulosic Biomass

Edwards

Henriksen

Yomano

et al. 2011

Appl Environ Microbiol

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“…Neither the chemical reduction nor the enzymatic oxidation of G 13 proceeded to completion; therefore, G 13 -R and G 13 -O were individually purified to homogeneity by semipreparative HPAEC-PAD (Spiro et al, 1993). Electrospray MS analysis confirmed that the expected product had been synthesized.…”

Section: Preparation Of Homogeneous Oligogalacturonides and Their Redmentioning

confidence: 99%

“…In this paper the term oligogalacturonide refers to native or unmodified, as well as reducing-end-derivatized oligogalacturonides, such as G 13 -T, G 13 -B, G 13 -R, and G 13 -O. Oligogalacturonides, generated by partial endopolygalacturonase digestion of polygalacturonic acid, were purified to homogeneity by a three-step protocol (Spiro et al, 1993) consisting of size-selective precipitation with ethanol, followed by Q-Sepharose anion-exchange chromatography, and then semipreparative HPAEC-PAD. All oligogalacturonides used in this study were homogeneous and chemically stable at Ϫ20°C, as determined by analytical HPAEC-PAD and by determination of their M r by electrospray MS.…”

Section: Preparation Of Homogeneous Oligogalacturonides and Their Redmentioning

confidence: 99%

Biological Activity of Reducing-End-Derivatized Oligogalacturonides in Tobacco Tissue Cultures1

et al. 1998

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The biological activity of reducing-end-modified oligogalacturonides was quantified in four tobacco (Nicotiana tabacum) tissue culture bioassays. The derivatives used were oligogalacturonides with the C-1 of their reducing end (a) covalently linked to a biotin hydrazide, (b) covalently linked to tyramine, (c) chemically reduced to a primary alcohol, or (d) enzymatically oxidized to a carboxylic acid. These derivatives were tested for their ability to (a) alter morphogenesis of N. tabacum cv Samsun thin cell-layer explants, (b) elicit extracellular alkalinization by suspension-cultured cv Samsun cells, (c) elicit extracellular alkalinization by suspensioncultured N. tabacum cv Xanthi cells, and (d) elicit H 2 O 2 accumulation in the cv Xanthi cells. In all four bioassays, each of the derivatives had reduced biological activity compared with the corresponding underivatized oligogalacturonides, demonstrating that the reducing end is a key element for the recognition of oligogalacturonides in these systems. However, the degree of reduction in biological activity depends on the tissue culture system used and on the nature of the specific reducing-end modification. These results suggest that oligogalacturonides are perceived differently in each tissue culture system.Carbohydrates that act as signal molecules in plants (oligosaccharins) have been isolated from plant and fungal cell wall polysaccharides, from the cell walls of bacterial symbionts of plants, and from fungal glycopeptides (for review, see Ryan and Farmer, 1991;Darvill et al., 1992;Cô té and Hahn, 1994). We are interested in determining the mechanism by which one type of the plant cell wallderived oligosaccharins, the oligogalacturonides, elicit biological responses in plants.The biological effects elicited in plants by oligosaccharins are diverse (for review, see Ryan and Farmer, 1991;Darvill et al., 1992;Cô té and Hahn, 1994) but can generally be placed in two groups: delayed responses and rapid responses. Delayed responses are usually observed hours or days after oligosaccharin treatment and are often directly involved in adaptation to environmental conditions, whereas rapid responses generally occur at the plant cell surface and are observed within minutes after addition of oligosaccharins. The delayed responses elicited by oligogalacturonides can be broadly divided into those in which defense responses are induced and those in which growth and development are modified. The defense-related responses, depending on the plant species, include phytoalexin accumulation (Hahn et al., 1981), lignification of cell walls (Robertsen, 1986), and proteinase inhibitor accumulation (Bishop et al., 1984). The responses involving growth and development include induction of ethylene in tomato fruit (Brecht and Huber, 1988), inhibition of auxin-induced pea stem elongation (Branca et al., 1988), and regulation of tobacco (Nicotiana tabacum) TCL explant morphogenesis (Eberhard et al., 1989). The rapid responses induced by oligogalacturonides include enhanced protein phosp...

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“…Sizehomogeneous OGs were prepared by high-performance anion-exchange chromatography (Dionex, Sunnyvale, CA) on a semipreparative CarboPac PA1 column (9 ϫ 250 mm) and analyzed by high-performance anion-exchange chromatography-pulsed-amperometric detection, as described by Spiro et al (1993). The DPs of the individual OGs were determined by comparison of their retention time with those of the standard OGs whose DPs had been determined by fast-atom-bombardment mass spectrometry (Marfà et al, 1991).…”

Section: Elicitor Preparationmentioning

confidence: 99%

Extracellular H2O2 Induced by Oligogalacturonides Is Not Involved in the Inhibition of the Auxin-Regulated rolB Gene Expression in Tobacco Leaf Explants

et al. 2000

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Purification and characterization of biologically active 1,4-linked α-d-oligogalacturonides after partial digestion of polygalacturonic acid with endopolygalacturonase

Cited by 73 publications

References 16 publications

Addition of Genes for Cellobiase and Pectinolytic Activity in Escherichia coli for Fuel Ethanol Production from Pectin-Rich Lignocellulosic Biomass

Addition of Genes for Cellobiase and Pectinolytic Activity in Escherichia coli for Fuel Ethanol Production from Pectin-Rich Lignocellulosic Biomass

Biological Activity of Reducing-End-Derivatized Oligogalacturonides in Tobacco Tissue Cultures1

Extracellular H2O2 Induced by Oligogalacturonides Is Not Involved in the Inhibition of the Auxin-Regulated rolB Gene Expression in Tobacco Leaf Explants

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