Purification and Properties of a Low-molecular-weight, High-alkaline Pectate Lyase from an Alkaliphilic Strain of<i>Bacillus</i>

Kobayashi, Tohru; Koike, Kenichi; Yoshimatsu, Tadashi; Higaki, Norihiko; Suzumatsu, Atsushi; Ozawa, Tadahiro; Hatada, Yuji; Ito, Susumu

doi:10.1271/bbb.63.65

Cited by 66 publications

(53 citation statements)

References 41 publications

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“…P-2850 have not been reported, pectate lyase from Bacillus sp. KSM-P15 shows a clear preference for polygalacturonic acid and very low activity on methylated pectins (Kobayashi et al, 1999). Despite their similarity, PelC and PelA show some differences regarding substrate specificity.…”

Section: Discussionmentioning

confidence: 99%

Pectate lyase C from Bacillus subtilis: a novel endo-cleaving enzyme with activity on highly methylated pectin

2006

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The gene yvpA from Bacillus subtilis was cloned and expressed in Escherichia coli. It encoded a pectate lyase of 221 amino acids that was denominated PelC. The heterologously expressed enzyme was purified by His-tag affinity chromatography and characterized. PelC depolymerized polygalacturonate and pectins of methyl esterification degree from 22 % to 89 %, exhibiting maximum activity on 22 % esterified citrus pectin. It showed an absolute Ca 2+ requirement and the optimum temperature and pH were 65 6C and pH 10, respectively. The deduced amino acid sequence of PelC showed 53 % identity to pectate lyase PelA from Paenibacillus barcinonensis, which was also characterized. Similarly to PelC, purified PelA showed activity on polygalacturonate and pectins with a high degree of methyl esterification. The two enzymes cleaved pectic polymers to a mixture of oligogalacturonates, indicating an endo mode of action. Analysis of activity on trigalacturonate showed that PelC cleaved it to galacturonic acid and unsaturated digalacturonate, whereas PelA did not show activity on this substrate. PelC and PelA showed high homology to a few recently identified pectate lyases of family 3 and form with them a cluster of small-sized pectate lyases from non-pathogenic micro-organisms.

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Section: Discussionmentioning

confidence: 99%

Pectate lyase C from Bacillus subtilis: a novel endo-cleaving enzyme with activity on highly methylated pectin

2006

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“…Different optimum activities have been reported for different pectinases from different sources. In the presence of Ca 2+ , the enzyme from strain of Bacillus spp KSM-P15 degraded polygalacturonic acid with an optimal activity around pH 10.5 and 50-55°C (Kobayashi et al, 1999). Namasivayam et al (2011) reported an optimum temperature for maximum activity of pectinase from B. cereus to be 37°C.…”

Section: Discussionmentioning

confidence: 99%

A comparative study of pectinolytic enzyme production by Bacillus species

Torimiro¹,

Okonji²

2013

Afr. J. Biotechnol.

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The economic and ecological function of pectinase enzymes in industries is gaining much attention with the need of highly productive strains of microorganisms to reduce production cost. The present investigation is a comparative evaluation of Bacillus sp. pectinases. The three isolates isolated from agro-waste, Bacillus stearothermophilus, Bacillus cereus and Bacillus subtilis, showed very high pectinase activities with pectin as the substrate. The three isolates also showed varying degree of preference for banana peels and wheat bran but their activity towards orange peel was very low compared to other substrate. The effect of temperature on the pectinase of the three isolates showed that B. stearothermophilus had optimum temperature at 60°C while B. cereus and B. subtilis both showed optimum activity at 50°C. On the effect of pH, B. stearothermophilus, B. cereus and B. subtilis showed optimum pectinase activities at pH 7.5, 8.0 and 9.0, respectively. Metal ions enhanced the activity of pectinase produced by the three isolates. The study exhibited that B. stearothermophilus gave the most pectinase activity, optimum temperature and a moderate alkaline pH, possibly making it a better source for industrial purpose.

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“…However, the sequence of the cloned enzyme shows high homology (54 % identity) to the protein deduced from the yvpA gene from B. subtilis (Kunst et al, 1997). Recently, a pectate lyase from an alkaliphilic strain of Bacillus has been characterized (Kobayashi et al, 1999). The enzyme shows similar molecular mass and pI (20 kDa and 10n3, respectively) to PelA, although it exhibits little activity on pectins.…”

Section: Discussionmentioning

confidence: 99%

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.

et al. 2000

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The gene pelA encoding a pectate lyase from the strain Bacillus sp. BP-23 was cloned and expressed in Escherichia coli. The nucleotide sequence of a 1214 bp DNA fragment containing pelA gene was determined, revealing an ORF of 666 nucleotides that encoded a protein of 23 233 Da. The deduced amino acid sequence of the encoded enzyme showed homology to pectate lyases A, B, C and D from Fusarium solani, Pel-3 and PelB from Erwinia carotovora and PelI from Erwinia chrysanthemi. Homology was also found to the protein deduced from the Bacillus subtilis yvpA gene, the function of which is unknown. The heterologous expressed enzyme depolymerized polygalacturonate and pectins of methyl esterification degree from 22 to 89 %, and exhibited similar activity on polygalacturonate and on 89 % esterified citrus pectin. Optimum temperature and pH for enzymic activity were 50 mC and pH 10, respectively. Ca 2M was required for activity on pectic substrates, while the enzyme was strongly inhibited by Ba 2M .

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Purification and Properties of a Low-molecular-weight, High-alkaline Pectate Lyase from an Alkaliphilic Strain ofBacillus

Cited by 66 publications

References 41 publications

Pectate lyase C from Bacillus subtilis: a novel endo-cleaving enzyme with activity on highly methylated pectin

Pectate lyase C from Bacillus subtilis: a novel endo-cleaving enzyme with activity on highly methylated pectin

A comparative study of pectinolytic enzyme production by Bacillus species

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.

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