Gene Cloning and Characterization of α-Glucuronidase of<i>Bacillus stearothermophilus</i>No. 236

Choi, In Keun; Kim, Hwa-Young; Choi, Yong-Jin

doi:10.1271/bbb.64.2530

Cited by 25 publications

(14 citation statements)

References 21 publications

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“…strain JDR-2 have been identified in several other bacteria (G. stearothermophilus, T. maritima, and Bacillus clausii). The enzymatic activity of AguA with MeGAX 3 as the substrate shares properties with the activities of other GH67 ␣-glucuronidases (4,7,21,27,29) and supports the unique role proposed for extracellular GH10 endoxylanases, in which MeGAX 3 is generated along with xylooligosaccharides, all of which are assimilated and metabolized (3,24).…”

Section: Discussionsupporting

confidence: 66%

Aldouronate Utilization in Paenibacillus sp. Strain JDR-2: Physiological and Enzymatic Evidence for Coupling of Extracellular Depolymerization and Intracellular Metabolism

Nong

Rice

Chow

et al. 2009

Appl Environ Microbiol

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Paenibacillus sp. strain JDR-2, an aggressively xylanolytic bacterium isolated from decaying sweet gum wood, secretes a multimodular glycohydrolase family GH10 endoxylanase (XynA1) anchored to the cell surface. The gene encoding XynA1 is part of a xylan utilization regulon that includes an aldouronate utilization gene cluster with genes encoding a GH67 ␣-glucuronidase (AguA), a GH10 endoxylanase (XynA2), and a GH43 arabinofuranosidase/␤-xylosidase (XynB). Here we show that this Paenibacillus sp. strain is able to utilize methylglucuronoxylose (MeGAX 1 ), an aldobiuronate product that accumulates during acid pretreatment of lignocellulosic biomass, and methylglucuronoxylotriose (MeGAX 3 ), the product of the extracellular XynA1 acting on methylglucuronoxylan (MeGAX n ). The average rates of utilization of MeGAX n , MeGAX 1 , and MeGAX 3 were 149.8, 59.4, and 54.3 g xylose equivalents ⅐ ml ؊1 ⅐ h ؊1 , respectively, and were proportional to the specific growth rates on the substrates. AguA was active with MeGAX 1 and MeGAX 3 , releasing 4-O-methyl-D-glucuronate ␣-1,2 linked to a nonreducing terminal xylose residue. XynA2 converted xylotriose, generated by the action of AguA on MeGAX 3 , to xylose and xylobiose. The ability to utilize MeGAX 1 provides a novel metabolic potential for bioconversion of acid hydrolysates of lignocellulosics. The 2.8-fold-greater rate of utilization of polymeric MeGAX n than that of MeGAX 3 indicates that there is coupling of extracellular depolymerization, assimilation, and intracellular metabolism, allowing utilization of lignocellulosics with minimal pretreatment. Along with adjacent genes encoding transcriptional regulators and ABC transporter proteins, the aguA and xynA2 genes in the cluster described above contribute to the efficient utilization of aldouronates derived from dilute acid and/or enzyme pretreatment protocols applied to the conversion of hemicellulose to biofuels and chemicals.

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Section: Discussionsupporting

confidence: 66%

Aldouronate Utilization in Paenibacillus sp. Strain JDR-2: Physiological and Enzymatic Evidence for Coupling of Extracellular Depolymerization and Intracellular Metabolism

Nong

Rice

Chow

et al. 2009

Appl Environ Microbiol

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“…5b). The pH optimum of TtAguA was found to be more similar to the -glucuronidases from A. niger (Kiryu et al 2005), G. stearothermophilus (Choi et al 2000), Phanerochaete chrysosporium (Castanares et al 1995), and Bacteroides J-37 (Kim et al 1997), which have an acidic pH optima of 6.5. Conversely, the -glucuronidase from T. maritima has an optimal pH of 7.8 (Suresh et al 2002).…”

Section: Biochemical Characterizationmentioning

confidence: 85%

Characterization of Thermotoga thermarum DSM 5069 α-Glucuronidase and Synergistic Degradation of Xylan

Wang

Shi

et al. 2016

BioResources

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* α-Glucuronidases are capable of breaking down the α-1,2-glycosidic bonds of 4-O-methyl-D-glucuronic acid residues. As an accessory enzyme, α-glucuronidase plays a vital role in xylan degradation. The recombinant α-glucuronidase from Thermotoga thermarum DSM 5069 was heterologously expressed in the Escherichia coli system, purified, and characterized. The purified enzyme exhibited optimal activity toward aldouronic acids at pH 6.5 and 80 °C. It was fairly thermostable and maintained 98% residual activity after incubation at 65 °C for 2.0 h. The kinetic parameters Km, Vmax, and kcat were 3.02 ± 0.16 mM, 88 ± 2 µmol min -1 mg

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“…It has already been proved that most bacterial α-glucuronidases, including C. polysaccharolyticus (158.0 kDa) [16], Bacillus stearothermophilus No. 236 (161.0 kDa) [17] and B.stearothermophilus T-6 (150.0 kDa) [18], consist of two subunits with Mw of around 75.0 kDa per subunit. In contrast, many fungal α-glucuronidases function as monomeric proteins with a higher Mw of 100.0 kDa per subunit due to the glycosylation [19].…”

Section: Resultsmentioning

confidence: 99%

“…Nevertheless, a majority of the α-glucuronidases only has the capacity to act on small model xylan or MeGlcA branched XOs [17], [20]. Thus, the more reducing sugar detected in enzyme-cocktail treatment was related to the synergistic action of Xyn10A and Agu67A on the different parts of xylan polymer or XOs.…”

Section: Resultsmentioning

confidence: 99%

Insight into Glycoside Hydrolases for Debranched Xylan Degradation from Extremely Thermophilic Bacterium Caldicellulosiruptor lactoaceticus

Jia

Wang

et al. 2014

PLoS ONE

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Caldicellulosiruptor lactoaceticus 6A, an anaerobic and extremely thermophilic bacterium, uses natural xylan as carbon source. The encoded genes of C. lactoaceticus 6A for glycoside hydrolase (GH) provide a platform for xylan degradation. The GH family 10 xylanase (Xyn10A) and GH67 α-glucuronidase (Agu67A) from C. lactoaceticus 6A were heterologously expressed, purified and characterized. Both Xyn10A and Agu67A are predicted as intracellular enzymes as no signal peptides identified. Xyn10A and Agu67A had molecular weight of 47.0 kDa and 80.0 kDa respectively as determined by SDS-PAGE, while both appeared as homodimer when analyzed by gel filtration. Xyn10A displayed the highest activity at 80°C and pH 6.5, as 75°C and pH 6.5 for Agu67A. Xyn10A had good stability at 75°C, 80°C, and pH 4.5–8.5, respectively, and was sensitive to various metal ions and reagents. Xyn10A possessed hydrolytic activity towards xylo-oligosaccharides (XOs) and beechwood xylan. At optimum conditions, the specific activity of Xyn10A was 44.6 IU/mg with beechwood xylan as substrate, and liberated branched XOs, xylobiose, and xylose. Agu67A was active on branched XOs with methyl-glucuronic acids (MeGlcA) sub-chains, and primarily generated XOs equivalents and MeGlcA. The specific activity of Agu67A was 1.3 IU/mg with aldobiouronic acid as substrate. The synergistic action of Xyn10A and Agu67A was observed with MeGlcA branched XOs and xylan as substrates, both backbone and branched chain of substrates were degraded, and liberated xylose, xylobiose, and MeGlcA. The synergism of Xyn10A and Agu67A provided not only a thermophilic method for natural xylan degradation, but also insight into the mechanisms for xylan utilization of C. lactoaceticus.

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Gene Cloning and Characterization of α-Glucuronidase ofBacillus stearothermophilusNo. 236

Cited by 25 publications

References 21 publications

Aldouronate Utilization in Paenibacillus sp. Strain JDR-2: Physiological and Enzymatic Evidence for Coupling of Extracellular Depolymerization and Intracellular Metabolism

Aldouronate Utilization in Paenibacillus sp. Strain JDR-2: Physiological and Enzymatic Evidence for Coupling of Extracellular Depolymerization and Intracellular Metabolism

Characterization of Thermotoga thermarum DSM 5069 α-Glucuronidase and Synergistic Degradation of Xylan

Insight into Glycoside Hydrolases for Debranched Xylan Degradation from Extremely Thermophilic Bacterium Caldicellulosiruptor lactoaceticus

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