An unusual pectate lyase from a Bacillus sp. with high activity on pectin: cloning and characterization The EMBL accession number for the nucleotide sequence determined in this work is AJ237980.

Soriano, Montserrat; Blanco, Ana Marín; Dı́az, Pilar; Pastor, F. I. Javier

doi:10.1099/00221287-146-1-89

Cited by 63 publications

(37 citation statements)

References 37 publications

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“…These results are in agreement with those on other pectate lyases published by Soriano et al (2000) and Boland et al (2010). Soriano et al (2000) studied pelA, from the strain Paenibacillus sp. BP-23, actives on pectins of any degree of esterification.…”

Section: Enzyme Characterisationsupporting

confidence: 83%

“…The majority of studies are regarding pectinases from Erwinia and Bacillus within the bacteria, pectinases from Aspergillus within fungi, and pectinases from Saccharomyces cerevisiae and Kluyveromyces marxianus within yeasts. Many pectic enzymes are recombinantly expressed in Escherichia coli (Soriano et al 2000); Wang et al 2011;Damak et al 2013).…”

Section: Introductionmentioning

confidence: 99%

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Identification and Characterisation of a Pectinolytic Enzyme from Paenibacillus xylanolyticus

Giacobbe¹,

Pepe²,

Ventorino³

et al. 2014

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Pectinolytic enzymes play an important role in the processing of lignocellulosic materials because of their ability to improve the access of cellulases to their substrate by removing pectins. The strain Paenibacillus xylanolyticus 2-6L3 was isolated from mature compost obtained from agroindustrial wastes, and the enzyme pectate lyase from P. xylanolyticus 2-6L3, named PaenxylPel, was partially purified and subjected to structural and functional characterisation. The enzyme exhibited an optimum temperature between 60 and 70 °C and optimal pH value of 9.0 for its pectinase activity on pectin from citrus fruit. PaenxylPel showed a thermoresistance and pH resistance higher than those of other pectate lyases so far described, with half-lives of 48 and 24 h at 60 and 70 °C, respectively, a retention of around 80% of activity after 96 h at 40 and 50 °C, and a half-life of about 15 days at pH 8.0. PaenxylPel followed Michaelis-Menten kinetics toward pectin from citrus fruit, pectin from sugar beet pulp, high-ester pectin extracted from citrus peel (> 50% esterified), and polygalacturonic acid (PLA). The ability to act on both PLA and highly methylated pectins, together with a double peak in the graph of optimum pH at pH 5 and 9, suggest that pectate lyase from P. xylanolyticus shows an unusual activity, combining traits of pectate lyase and pectin lyase. This is the first manuscript on the pectinolytic activity of P. xylanolyticus.

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Section: Enzyme Characterisationsupporting

confidence: 83%

Section: Introductionmentioning

confidence: 99%

Identification and Characterisation of a Pectinolytic Enzyme from Paenibacillus xylanolyticus

Giacobbe¹,

Pepe²,

Ventorino³

et al. 2014

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“…Several pectate lyases from mesophilic bacteria, above all from members of the genus Erwinia but also from the gram-positive genus Bacillus, have been cloned and characterized (Pissavin et al 1996;Tardy et al 1997;Shevchik et al 1997;Bauer and Collmer 1997;Soriano et al 2000). Recently, a thermostable pectate lyase from a newly isolated thermophilic bacterium, Thermoanaerobacter italicus, has been isolated and characterized (Kozianowski et al 1997).…”

Section: Discussionmentioning

confidence: 99%

Cloning of two pectate lyase genes from the marine Antarctic bacterium Pseudoalteromonas haloplanktis strain ANT/505 and characterization of the enzymes

et al. 2001

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A marine Antarctic psychrotolerant bacterium (strain ANT/505), isolated from sea ice-covered surface water from the Southern Ocean, showed pectinolytic activity on citrus pectin agar. The sequencing of the 16S rRNA of isolate ANT/505 indicates a taxonomic affiliation to Pseudoalteromonas haloplanktis . The supernatant of this strain showed three different pectinolytic activities after growth on citrus pectin. By activity screening of a genomic DNA library of isolate ANT/505 in Escherichia coli , two different pectinolytic clones could be isolated. Subcloning and sequencing revealed two open reading frames (ORF) of 1,671 and 1,968 nt, corresponding to proteins of 68 and 75 kDa, respectively. The deduced amino acid sequence of the two ORFs showed homology to pectate lyases from Erwinia chrysanthemi and Aspergillus nidulans . The pectate lyases contain signal peptides of 17 and 26 amino acids that were correctly processed after overexpression in E. coli BL21. Both enzymes were purified by anionic exchange chromatography. Maximal enzymatic activities for both pectate lyases were observed at 30 ° C and a pH range of 9 to 10. The K m values of both lyases for pectate and citrus pectin were 1 g l -1 and 5 g l -1, respectively. Calcium was required for activity on pectic substrates, whereas the addition of 1 mM ethylenediaminetetraacetic acid (EDTA) resulted in complete inhibition of the enzymes. These two enzymes represent the first pectate lyases isolated and characterized from a cold-adapted marine bacterium.

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“…BP-23 (formerly Bacillus sp. BP-23) [19] shows a multienzymatic glycanase system, including several cellulases [20,21], xylanases [22,23], or pectinases [24]. In this study we report the cloning, purification and characterization of Paenibacillus sp.…”

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confidence: 99%

Exo‐mode of action of cellobiohydrolase Cel48C from Paenibacillus sp. BP‐23

Sánchez

Pastor

Dı́az

2003

European Journal of Biochemistry

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Sequence analysis of a Paenibacillus sp. BP-23 recombinant clone coding for a previously described endoglucanase revealed the presence of an additional truncated ORF with homology to family 48 glycosyl hydrolases. The corresponding 3509-bp DNA fragment was isolated after gene walking and cloned in Escherichia coli Xl1-Blue for expression and purification. The encoded enzyme, a cellulase of 1091 amino acids with a deduced molecular mass of 118 kDa and a pI of 4.85, displayed a multidomain organization bearing a canonical family 48 catalytic domain, a bacterial type 3a cellulose-binding module, and a putative fibronectin-III domain. The cloned cellulase, unique among Bacillales and designated Cel48C, was purified through affinity chromatography using its ability to bind Avicel. Maximum activity was achieved at 45°C and pH 6.0 on acid-swollen cellulose, bacterial microcrystalline cellulose, Avicel and cellodextrins, whereas no activity was found on carboxy methyl cellulose, cellobiose, cellotriose, pNPglycosides or 4-methylumbeliferyl a-D-glucoside. Cellobiose was the major product of cellulose hydrolysis, identifying Cel48C as a processive cellobiohydrolase. Although no chromogenic activity was detected from pNP-glycosides, TLC analysis revealed the release of p-nitrophenyl-glycosides and cellodextrins from these substrates, suggesting that Cel48C acts from the reducing ends of the sugar chain. Presence of such a cellobiohydrolase in Paenibacillus sp. BP-23 would contribute to widen up its range of action on natural cellulosic substrates.

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An unusual pectate lyase from a Bacillus sp. with high activity on pectin: cloning and characterization The EMBL accession number for the nucleotide sequence determined in this work is AJ237980.

Cited by 63 publications

References 37 publications

Identification and Characterisation of a Pectinolytic Enzyme from Paenibacillus xylanolyticus

Identification and Characterisation of a Pectinolytic Enzyme from Paenibacillus xylanolyticus

Cloning of two pectate lyase genes from the marine Antarctic bacterium Pseudoalteromonas haloplanktis strain ANT/505 and characterization of the enzymes

Exo‐mode of action of cellobiohydrolase Cel48C from Paenibacillus sp. BP‐23

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