Purification and characterization of pectinesterase produced by a strain ofAspergillus niger

Maldonado, María Cristina; Saad, A. M. Strasser De; Callieri, D. A. S.

doi:10.1007/bf01575960

Cited by 24 publications

(24 citation statements)

References 10 publications

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“…At pH 7 and above in the presence of 100 mM NaCl, Ani-PME2 is nearly inactive. A similar activity profile has been established for Ani-PME1 (79,100).…”

Section: Resultsmentioning

confidence: 59%

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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“…At pH 7 and above in the presence of 100 mM NaCl, Ani-PME2 is nearly inactive. A similar activity profile has been established for Ani-PME1 (79,100).…”

Section: Resultsmentioning

confidence: 59%

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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“…Table 4 showed that PE production slightly increased at about 74 h of fermentation. At pH higher than 5.0, PE production increased with pectin since the synthesis of this enzyme was induced by this substrate (Maldonado and Callieri, 1989;Maldonado et al, 1994).…”

Section: Resultsmentioning

confidence: 96%

Production of pectinases by A. niger: influence of fermentation conditions

Martos

Vázquez

Benassi

et al. 2009

Braz. arch. biol. technol.

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Response surface methodology was used for optimization of polygalacturonase (PG) and pectinesterase (PE) production in submerged fermentation by A.niger. A Central Composite Experimental Design was applied, consisting of 22 experiments, including eight central points. Variables studied were: fermentation time (24 to 120 h), pH (3.5 to 6.5) and initial concentration of pectin (5 to 20 g/l). Maximum PE production was 220 U/l, after 74 h of culture, in a medium containing 20 g/l of pectin (pH 6.5). The optimal conditions for PG production were pH: 4.1, 20 g/l of pectin and 94 h of fermentation with a maximum value of 1032 U/l. Under these conditions, the PE production was low (15 U/l). A liquid extract with high PG activity and low PE activity could be suitable to be used in food processing in order to reduce the production of methanol.

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“…Previous studies revealed that PMEs was a highly specific enzyme for the D-galacturonan structure and its activity was affected by several factors such as pH, ionic strength and temperature [39]. For example, Aspergillus niger PMEs had an optimal pH of 5 for enzymatic activity [40], whereas the pH optimum for P. capsici PMEs was 6.5 [10]. In addition, pectin and pectic acid in plant hosts induced microorganisms to produce PME.…”

Section: Discussionmentioning

confidence: 98%

Isolation of nine Phytophthora capsici pectin methylesterase genes which are differentially expressed in various plant species

Li¹,

Feng²,

Wang³

et al. 2011

J. Basic Microbiol.

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Phytophthora capsici causes damage on many plants species, and secretes various pectin methylesterases during all stages of infection. We identified nine Pme genes (Pcpme 1-9) from a genomic library of highly virulent P. capsici strain SD33 and further analyzed the expression pattern of nine genes on three hosts: pepper, tomato, and cucumber using qRT-PCR during all stages of infection. All nine genes were found to be differentially expressed in three host species in the course of P. capsici interaction. The expression levels of the respective genes increased from 1 to 7 dpi in pepper, while most genes presented a decreasing trend of expression from 1 to 5 dpi in tomato fruits. However, in both fruits peaks were reached at 7 dpi. In cucumber fruits, each gene showed minor expression levels from 1 to 3 dpi, exhibited definite peaks at 5 dpi, and then decreased from 5 to 7 dpi. Thus, evidence from our studies of Pcpme gene expression in different plants at a rang of time points suggests that the late stages of infection may represent the most critical time for P. capsici to successfully express or/and secret PMEs.

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Purification and characterization of pectinesterase produced by a strain ofAspergillus niger

Cited by 24 publications

References 10 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

Production of pectinases by A. niger: influence of fermentation conditions

Isolation of nine Phytophthora capsici pectin methylesterase genes which are differentially expressed in various plant species

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