Purification and partial characterisation of pectin methylesterase produced by Fusarium asiaticum

Glinka, E. M.; Liao, Yu-Cai

doi:10.1016/j.funbio.2011.07.005

Cited by 15 publications

(6 citation statements)

References 34 publications

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“…Although most fruits are acidic and hence a PME operating at acidic pH, such as Ani-PME2, is optimal for pectin hydrolysis, seeds and bulbs generally have a pH value close to neutral. A 31-kDa PME (one of three) from the fungus Fusarium asiaticum has been found to have a pH optimum of 6.5 (104), and there is a third putative PME for A. niger that awaits characterization.…”

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confidence: 99%

Structure and Properties of a Non-processive, Salt-requiring, and Acidophilic Pectin Methylesterase from Aspergillus niger Provide Insights into the Key Determinants of Processivity Control

Kent

Loo

Melton

et al. 2016

Journal of Biological Chemistry

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Many pectin methylesterases (PMEs) are expressed in plants to modify plant cell-wall pectins for various physiological roles. These pectins are also attacked by PMEs from phytopathogens and phytophagous insects. The de-methylesterification by PMEs of the O6-methyl ester groups of the homogalacturonan component of pectin, exposing galacturonic acids, can occur processively or non-processively, respectively, describing sequential versus single de-methylesterification events occurring before enzyme-substrate dissociation. The high resolution x-ray structures of a PME from Aspergillus niger in deglycosylated and Asn-linked N-acetylglucosamine-stub forms reveal a 10 2 ⁄ 3-turn parallel ␤-helix (similar to but with less extensive loops than bacterial, plant, and insect PMEs). Capillary electrophoresis shows that this PME is non-processive, halophilic, and acidophilic. Molecular dynamics simulations and electrostatic potential calculations reveal very different behavior and properties compared with processive PMEs. Specifically, uncorrelated rotations are observed about the glycosidic bonds of a partially de-methyl-esterified decasaccharide model substrate, in sharp contrast to the correlated rotations of processive PMEs, and the substrate-binding groove is negatively not positively charged.The heterogeneous polysaccharide pectin (1-4) is a key component of the plant cell wall (5). A host of enzymes, including pectin methylesterase (PME) 3 as well as endo-and exopolygalacturonases, modifies pectin fine-structure locally in space and time for purposes of plant-cell differentiation, cell adhesion/separation, growth, and development from roots to meristem, morphogenesis, seed and fruit development, and defense against pathogens (6 -16). PMEs hydrolyze the O6-methyl ester groups of the homogalacturonan (HG) chains that form the backbone of pectin (Fig. 1). As well as being ubiquitous in plants, and frequently manifesting in a staggering number of isoforms (more than 66 for Arabidopsis thaliana) (8, 10), PMEs are found in phytopathogenic bacteria and fungi (17-26) and in Archaea (especially Halobacteriaceae (27)), and they have also been observed in the genomes of several phytophagous insects (28 -30) and in human gut microflora (31). Recently, plant pollen PMEs have been identified as a key human allergen (32, 33). Bacterial and plant PMEs that have been functionally characterized are generally observed to be processive (34 -55), i.e. the enzyme binds to the HG substrate and then successively hydrolyzes a block of methyl ester groups before dissociating. In all cases characterized to date, the direction of processivity is from the non-reducing end of the HG chain toward the reducing end, as shown in Fig. 1. The degree of processivity (also called the action pattern) has been reported to be dependent on ionic strength and pH (56), and various classifications of PMEs exist (48,49,(57)(58)(59)(60). In general, processive PMEs have their isoelectric points, pI, and pH for optimum activity at near-neutral to basic pH, although ...

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Section: )mentioning

confidence: 99%

Structure and Properties of a Non-processive, Salt-requiring, and Acidophilic Pectin Methylesterase from Aspergillus niger Provide Insights into the Key Determinants of Processivity Control

Kent

Loo

Melton

et al. 2016

Journal of Biological Chemistry

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“…Though different forms of other pectinases have been reported from Fusarium moniliformae , Fusarium oxysporum f sp. lycoperisi , Fusarium asiaticum , and Fusarium solani in submerged cultures, the reports of PNLs from Fusarium genera are very few . Keeping the above points in view, authors have reported purification and characterization of an alkaline PNL from a new fungal strain Fusarium decemcellulare MTCC 2079 under solid state fermentation culture conditions and have demonstrated the use of this enzyme in the retting of Crotolaria juncea fibers.…”

Section: Introductionmentioning

confidence: 99%

Purification and biochemical characterization of an alkaline pectin lyase from Fusarium decemcellulareMTCC 2079 suitable for Crotolaria juncea fiber retting

Yadav

Dubey

Anand

et al. 2013

J. Basic Microbiol.

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An extracellular pectin lyase secreted by Fusarium decemcellulare MTCC 2079 under solid state fermentation condition has been purified to electrophoretic homogeniety by using ammonium sulfate fractionation, carboxymethyl cellulose and gel filtration (Sephadex G-100) column chromatographies. The purified enzyme showed single protein band corresponding to molecular mass 45 ± 01 kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme had maximum activity at pH 9.0 and showed maximum stability in the pH range of 9.0-12.0. The optimum temperature of the purified enzyme was 50 °C and it showed maximum stability upto 40 °C. The energy of activation for the thermal denaturation (Ea ) was 59.06 kJ mol(-1) K(-1). The Km and kcat values using citrus pectin as the substrate were 0.125 mg ml(-1) and 72.9 s(-1) in 100 mM sodium carbonate buffer pH 9.0 at 50 °C. The biophysical studies on pectin lyase showed that its secondary structure belongs to α + β class of protein with comparatively less of β-sheets. Purified pectin lyase showed efficient retting of Crotolaria juncea fibers.

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“…Robl et al (2013) analyzed the enzymatic production of hemicelullases and related enzymes, with use of agar aesculin and agar with specific substrate evaluating the halo degradation for the initial selection. Glinka and Liao (2011) analyzed the production of pectin metilesterase (PME), polygalacturonase (PG) and pectin lyase (PL) through activity tests in the agar diffusion assays. Downie et al (1998) also examined the quantification of pectin metilesterase activity by agar diffusion method.…”

Section: Resultsmentioning

confidence: 99%

Rapid microbiological tests for prospecting new fungal strains with high potentiality for the pectinolytic enzymes production

Cangussu¹,

Felix²,

Araujo³

et al. 2016

Afr. J. Microbiol. Res.

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Prospecting of new isolates producers of pectinolitic enzymes were performed with cultivation of substrates of rice, soybean and corn bran. Pectinolitic enzymes were observed in cultures of the isolates NFC 1, NFC 2, NFC 4, NFC 5 and NFC 6 identified as belonging to the genus Aspergillus sp. Isolates NFR 1, NFR 2 and NFS1 were identified as belonging to the genus Rhizopus and NFC 3 belonging to the genus Cladosporium; however, pectinolitic enzymes production was not observed. The polygalacturonase (PG) production in submerged fermentation of Aspergillus NFC 2 in culture medium proposed (SM) at 28 and 37°C was quantified by agar diffusion and spectrophotometric methods. The highest PG production was obtained at 28°C by greater formation of halo degradation pectin and the activity measured by spectrophotometric methods. The activity of PG obtained was 7 U.mL -1 with volumetric productivity greater in the first 24 h of fermentation both at 28 and 37°C. A decrease in PG synthesis occurred after 48 to 120 h. The study allowed, by means of rapid microbiological tests, the selection of new isolates fungi for PG production and the definition of crucial stages for cultivation in liquid medium.

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Purification and partial characterisation of pectin methylesterase produced by Fusarium asiaticum

Cited by 15 publications

References 34 publications

Structure and Properties of a Non-processive, Salt-requiring, and Acidophilic Pectin Methylesterase from Aspergillus niger Provide Insights into the Key Determinants of Processivity Control

Structure and Properties of a Non-processive, Salt-requiring, and Acidophilic Pectin Methylesterase from Aspergillus niger Provide Insights into the Key Determinants of Processivity Control

Purification and biochemical characterization of an alkaline pectin lyase from Fusarium decemcellulareMTCC 2079 suitable for Crotolaria juncea fiber retting

Rapid microbiological tests for prospecting new fungal strains with high potentiality for the pectinolytic enzymes production

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