Thermostable Xylanase from Marasmius sp.: Purification and Characterization

Ratanachomsri, Ukrit; Sriprang, Rutchadaporn; Sornlek, Warasirin; Buaban, Benchaporn; Champreda, Verawat; Tanapongpipat, Sutipa; Eurwilaichitr, Lily

doi:10.5483/bmbrep.2006.39.1.105

Cited by 18 publications

(14 citation statements)

References 16 publications

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“…1). The molecular masses (24 kDa) of the enzymes are similar to the values of 23.6 and 25.6 kDa obtained for Scopulariopsis xylanase (Afzal et al 2005) but different from those of others (Ratanachomsri et al 2006).…”

Section: Purification Of Xylanasesmentioning

confidence: 71%

Purification and characterization of xylanases from Thermomyces lanuginosus and its mutant derivative possessing novel kinetic and thermodynamic properties

Bokhari

Latif

Rajoka

2008

World J Microbiol Biotechnol

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Xylanases produced from a locally isolated strain of Thermomyces lanuginosus and its mutant derivative were purified to a yield of 39.1 and 42.83% with specific activities of 15,501 and 17,778 IU mg -1 protein, respectively. The purification consisted of two steps i.e., ammonium sulphate precipitation, and gel filtration chromatography. The mutant enzyme showed high affinity for substrate, with a K m of 0.098 mg ml -1 as compared to wild type enzyme showing K m of not less than 0.112 mg ml -1 . It was found that pH values of 8.1 and 7.3 were best for activity of the mutant and wild-type-derived enzymes, respectively. The values of pK a of the acidic limbs of both enzymes were the same (5.0 and 4.9, respectively) but the pK a value of the basic limb was slightly increased, indicating the participation of a carboxyl group present in a non-polar environment. Temperatures of 70 and 65°C were found optimal for mutant and wild-derived xylanase, respectively. Enzymes displayed a high thermostability showing a half life of 31.79 and 6.0 min (5.3-fold improvement), enthalpy of denaturation (DH*) of 146.06 and 166.95 kJ mol -1 , entropy of denaturation (DS*) of 101.44 and 174.67, and free energy of denaturation (DG*) of 110.25 and 105.29 kJ mol -1 for mutant-and wildorganism derived enzyme, respectively at 80°C. Studies on the folding and stability of cellulase-less xylanases are important, since their biotechnological employments require them to function under extreme conditions of pH and temperature. The kinetic and thermodynamic properties suggested that the xylanase from the mutant organism is better as compared to xylanase produced from the wild type and previously reported strains of same species, and may have a potential usage in various industrial fields.

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Section: Purification Of Xylanasesmentioning

confidence: 71%

Purification and characterization of xylanases from Thermomyces lanuginosus and its mutant derivative possessing novel kinetic and thermodynamic properties

Bokhari

Latif

Rajoka

2008

World J Microbiol Biotechnol

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“…Relatively few xylanases described to date have temperature optima of Ն90°C; most of these were isolated from hyperthermophiles (growth T opt Ͼ 85°C) such as Thermotoga (27,37) or from high-temperature environments where hyperthermophiles are found (32). Another exception is the hightemperature xylanase (T opt ϭ 90°C) from a fungus (24).…”

mentioning

confidence: 99%

“…It should be noted that Xyn10A is actively degrading xylan during the incubation at 90°C, and loss of activity after longer incubation periods may be due to the depletion of the stabilizing substrate. Certain other xylanases are known to be stabilized by substrate or by immobilization on glass beads at high temperature (24,27), and in some cases, carbohydrate-binding domains were shown to contribute to thermostability (2,11,33,36). Xyn10A lacks predicted carbohydrate-binding domains, and therefore how Xyn10A interacts with xylans for stabilization is unknown.…”

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confidence: 99%

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Xyn10A, a Thermostable Endoxylanase from Acidothermus cellulolyticus 11B

Barabote¹,

Parales

Guo³

et al. 2010

Appl Environ Microbiol

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We cloned and purified the major family 10 xylanase (Xyn10A) from Acidothermus cellulolyticus 11B. Xyn10A was active on oat spelt and birchwood xylans between 60°C and 100°C and between pH 4 and pH 8. The optimal activity was at 90°C and pH 6; specific activity and K m for oat spelt xylan were 350 mol xylose produced min ؊1 mg of protein ؊1 and 0.53 mg ml ؊1 , respectively. Based on xylan cleavage patterns, Xyn10A is an endoxylanase, and its half-life at 90°C was approximately 1.5 h in the presence of xylan.

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“…The natural diversity of enzymes provides these industries with candidates having bifunctional activity, such as the xylanases from Aspergillus niger (12) and Marasmius sp. (26) that have xylanase and cellulase activities and the bifunctional xylanase-lichenase from Ruminococcus flavefaciens 17 (7). On the other hand, many artificial bifunctional xylanases have been synthesized for more efficient biodegradation of plant fiber (19).…”

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confidence: 99%

Paenibacillus sp. Strain E18 Bifunctional Xylanase-Glucanase with a Single Catalytic Domain

Shi

Tian

Yuan

et al. 2010

Appl Environ Microbiol

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Xylanases are utilized in a variety of industries for the breakdown of plant materials. Most native and engineered bifunctional/multifunctional xylanases have separate catalytic domains within the same polypeptide chain. Here we report a new bifunctional xylanase (XynBE18) produced by Paenibacillus sp. E18 with xylanase and ␤-1,3-1,4-glucanase activities derived from the same active center by substrate competition assays and site-directed mutagenesis of xylanase catalytic Glu residues (E129A and E236A). The gene consists of 981 bp, encodes 327 amino acids, and comprises only one catalytic domain that is highly homologous to the glycoside hydrolase family 10 xylanase catalytic domain. Recombinant XynBE18 purified from Escherichia coli BL21(DE3) showed specificity toward oat spelt xylan and birchwood xylan and ␤-1,3-1,4-glucan (barley ␤-glucan and lichenin). Homology modeling and molecular dynamic simulation were used to explore structure differences between XynBE18 and the monofunctional xylanase XynE2, which has enzymatic properties similar to those of XynBE18 but does not hydrolyze ␤-1,3-1,4-glucan. The cleft containing the active site of XynBE18 is larger than that of XynE2, suggesting that XynBE18 is able to bind larger substrates such as barley ␤-glucan and lichenin. Further molecular docking studies revealed that XynBE18 can accommodate xylan and ␤-1,3-1,4-glucan, but XynE2 is only accessible to xylan. These results indicate a previously unidentified structurefunction relationship for substrate specificities among family 10 xylanases.

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Thermostable Xylanase from Marasmius sp.: Purification and Characterization

Cited by 18 publications

References 16 publications

Purification and characterization of xylanases from Thermomyces lanuginosus and its mutant derivative possessing novel kinetic and thermodynamic properties

Purification and characterization of xylanases from Thermomyces lanuginosus and its mutant derivative possessing novel kinetic and thermodynamic properties

Xyn10A, a Thermostable Endoxylanase from Acidothermus cellulolyticus 11B

Paenibacillus sp. Strain E18 Bifunctional Xylanase-Glucanase with a Single Catalytic Domain

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