Molecular Cloning and Expression in<i>Pichia pastoris</i>of a<i>Irpex lacteus</i>Exo-β-(1→3)-galactanase Gene

Kotake, Toshihisa; Kitazawa, Kiminari; Takata, Ryohei; Okabe, Kohei; Ichinose, Hitomi; Kaneko, Satoshi; Tsumuraya, Yoichi

doi:10.1271/bbb.90433

Cited by 33 publications

(41 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…With regard to the arabinogalactanase previously found in R. niveus, we could not find definitive information about its mode of action (24 -26). The enzyme has been found to hydrolyze coffee AG into ␤-1,6-Gal n ; however, these oligosaccharides could also be released by the action of exo-␤-1,3-galactanase (22,43). We could not find a sequence related to FvEn3GAL in the genomic database of an organism of the Rhizopus genus, Rhizopus oryzae (Rhizopus oryzae Database available from the Broad Institute).…”

Section: Discussionmentioning

confidence: 86%

“…These observations suggest that the ␤-1,6-galactosyl branches of AG moieties hinder the access of the enzyme to the ␤-1,3-galactan main chains. Supporting this suggestion is the fact that removal of ␤-1,6-galactosyl branches from gum arabic by Smith degradation made it more susceptible to the enzyme, showing that ␤-galactan II with fewer ␤-1,6-galactosyl branches is a better substrate than gum arabic and ␤-galactan I (Table 1) (22,43). It seems likely that FvEn3GAL acted only on the limited parts of ␤-1,3-galactan main chains devoid of branches in the AGPs.…”

Section: Purification Of the Enzyme From The Culture Medium Of F Velmentioning

confidence: 97%

See 1 more Smart Citation

Endo-β-1,3-galactanase from Winter Mushroom Flammulina velutipes

Kotake

Hirata

Degi

et al. 2011

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Arabinogalactan proteins are proteoglycans found on the cell surface and in the cell walls of higher plants. The carbohydrate moieties of most arabinogalactan proteins are composed of ␤-1,3-galactan main chains and ␤-1,6-galactan side chains, to which other auxiliary sugars are attached. For the present study, an endo-␤-1,3-galactanase, designated FvEn3GAL, was first purified and cloned from winter mushroom Flammulina velutipes. The enzyme specifically hydrolyzed ␤-1,3-galactan, but did not act on ␤-1,3-glucan, ␤-1,3:1,4-glucan, xyloglucan, and agarose. It released various ␤-1,3-galactooligosaccharides together with Gal from ␤-1,3-galactohexaose in the early phase of the reaction, demonstrating that it acts on ␤-1,3-galactan in an endo-fashion. Phylogenetic analysis revealed that FvEn3GAL is member of a novel subgroup distinct from known glycoside hydrolases such as endo-␤-1,3-glucanase and endo-␤-1,3:1,4-glucanase in glycoside hydrolase family 16. Point mutations replacing the putative catalytic Glu residues conserved for enzymes in this family with Asp abolished activity. These results indicate that FvEn3GAL is a highly specific glycoside hydrolase 16 endo-␤-1,3-galactanase.

show abstract

Section: Discussionmentioning

confidence: 86%

Section: Purification Of the Enzyme From The Culture Medium Of F Velmentioning

confidence: 97%

Endo-β-1,3-galactanase from Winter Mushroom Flammulina velutipes

Kotake

Hirata

Degi

et al. 2011

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…To date, all characterized exo-␤-1,3-galactanases have greater activity toward ␤-1,3-galactan than ␤-1,3/␤-1,6-galactan from Prototheca zopfii, LWAG, gum arabic, and radish roots (1,(3)(4)(5)(6)(7)21). BLLJ_1840 has low sequence identities (27% to 28%) with other exo-␤-1,3-galactanases.…”

Section: Discussionmentioning

confidence: 99%

“…In particular, exo-␤-1,3-galactanase hydrolyzes the ␤-1,3-galactan backbone, bypassing ␤-1,6-galactan side chains, and consequently releases galactose, ␤-1,6-galactooligosaccharides, and their derivatives (1,2). Exo-␤-1,3-galactanases, which belong to glycoside hydrolase family 43 (GH43), have been cloned and characterized from several sources, including bacteria and fungi (1,(3)(4)(5)(6)(7).…”

mentioning

confidence: 99%

Bifidobacterium longum subsp. longum Exo-β-1,3-Galactanase, an Enzyme for the Degradation of Type II Arabinogalactan

Fujita

Sakaguchi

Sakamoto

et al. 2014

Appl Environ Microbiol

View full text Add to dashboard Cite

Type II arabinogalactan (AG-II) is a suitable carbohydrate source for Bifidobacterium longum subsp. longum, but the degradative enzymes have never been characterized. In this study, we characterized an exo-␤-1,3-galactanase, BLLJ_1840, belonging to glycoside hydrolase family 43 from B. longum subsp. longum JCM1217. The recombinant BLLJ_1840 expressed in Escherichia coli hydrolyzed ␤-1,3-linked galactooligosaccharides but not ␤-1,4-and ␤-1,6-linked galactooligosaccharides. The enzyme also hydrolyzed larch wood arabinogalactan (LWAG), which comprises a ␤-1,3-linked galactan backbone with ␤-1,6-linked galactan side chains. The k cat /K m ratio of dearabinosylated LWAG was 24-fold higher than that of ␤-1,3-galactan. BLLJ_1840 is a novel type of exo-␤-1,3-galactanase with a higher affinity for the ␤-1,6-substituted ␤-1,3-galactan than for nonsubstituted ␤-1,3-galactan. BLLJ_1840 has 27% to 28% identities with other characterized exo-␤-1,3-galactanases from bacteria and fungi. The homologous genes are conserved in several strains of B. longum subsp. longum and B. longum subsp. infantis but not in other bifidobacteria. Transcriptional analysis revealed that BLLJ_1840 is intensively induced with BLLJ_1841, an endo-␤-1,6-galactanase candidate, in the presence of LWAG. This is the first report of exo-␤-1,3-galactanase in bifidobacteria, which is an enzyme used for the acquisition of AG-II in B. longum subsp. longum.

show abstract

“…The N-terminally and C-terminally located modules appear to derive from cluster A and cluster C of our phylogenetic tree, respectively. It is (46), BAD89094 (Penicillium chrysogenum 31B), BAD15018 (Penicillium chrysogenum 31B) (47), BAE55732 (Aspergillus oryzae RIB40) (56), AAC67554 (Cochliobolus carbonum) (71), XP_391644 (Gibberella zeae PH-1), BAC75546 (Penicillium herquei IFO 4674) (24), XP_391670 (Gibberella zeae PH-1), CAL81199 (Humicola insolens DSM 18000) (54), ACP50519 (Penicillium purpurogenum MYA-38), BAH29957 (Irpex lacteus NBRC5367) (32), and BAD98241 (Phanerochaete chrysosporium) (22). The identifications of the proteins from F. succinogenes S85 were as denoted in the FibRumba database (http://www.jcvi.org/rumenomics).…”

Section: Discussionmentioning

confidence: 99%

Domain Analysis of a Modular α- l -Arabinofuranosidase with a Unique Carbohydrate Binding Strategy from the Fiber-Degrading Bacterium Fibrobacter succinogenes S85

et al. 2010

View full text Add to dashboard Cite

Family 43 glycoside hydrolases (GH43s) are known to exhibit various activities involved in hemicellulose hydrolysis. Thus, these enzymes contribute to efficient plant cell wall degradation, a topic of much interest for biofuel production. In this study, we characterized a unique GH43 protein from Fibrobacter succinogenes S85. The recombinant protein showed ␣-L-arabinofuranosidase activity, specifically with arabinoxylan. The enzyme is, therefore, an arabinoxylan arabinofuranohydrolase (AXH). The F. succinogenes AXH (FSUAXH1) is a modular protein that is composed of a signal peptide, a GH43 catalytic module, a unique ␤-sandwich module (XX domain), a family 6 carbohydrate-binding module (CBM6), and F. succinogenes-specific paralogous module 1 (FPm-1). Truncational analysis and site-directed mutagenesis of the protein revealed that the GH43 domain/XX domain constitute a new form of carbohydrate-binding module and that residue Y484 in the XX domain is essential for binding to arabinoxylan, although protein structural analyses may be required to confirm some of the observations. Kinetic studies demonstrated that the Y484A mutation leads to a higher k cat for a truncated derivative of FSUAXH1 composed of only the GH43 catalytic module and the XX domain. However, an increase in the K m for arabinoxylan led to a 3-fold decrease in catalytic efficiency. Based on the knowledge that most XX domains are found only in GH43 proteins, the evolutionary relationships within the GH43 family were investigated. These analyses showed that in GH43 members with a XX domain, the two modules have coevolved and that the length of a loop within the XX domain may serve as an important determinant of substrate specificity.

show abstract

Molecular Cloning and Expression inPichia pastorisof aIrpex lacteusExo-β-(1→3)-galactanase Gene

Cited by 33 publications

References 28 publications

Endo-β-1,3-galactanase from Winter Mushroom Flammulina velutipes

Endo-β-1,3-galactanase from Winter Mushroom Flammulina velutipes

Bifidobacterium longum subsp. longum Exo-β-1,3-Galactanase, an Enzyme for the Degradation of Type II Arabinogalactan

Domain Analysis of a Modular α- l -Arabinofuranosidase with a Unique Carbohydrate Binding Strategy from the Fiber-Degrading Bacterium Fibrobacter succinogenes S85

Contact Info

Product

Resources

About