RESUMO
Xylanase activity was isolated from crude extracts of Trichoderma harzianum strains C and 4 grown at 28 degree C in a solid medium containing wheat bran as the carbon source. Enzyme activity was demonstrable in the permeate after ultrafiltration of the crude extracts using an Amicon system. The hydrolysis patterns of different xylans and paper pulps by xylanase activity ranged from xylose, xylobiose and xylotriose to higher xylooligosaccharides. A purified ss-xylosidase from the Trichoderma harzianum strain released xylose, xylobiose and xylotriose from seaweed, deacetylated, oat spelt and birchwood xylans. The purified enzyme was not active against acetylated xylan and catalyzed the hydrolysis of xylooligosaccharides, including xylotriose, xylotetraose and xylopentaose. However, the enzyme was not able to degrade xylohexaose. Xylanase pretreatment was effective for hardwood kraft pulp bleaching. Hardwood kraft pulp bleached in the XEOP sequence had its kappa number reduced from 13.2 to 8.9 and a viscosity of 20. 45 cp. The efficiency of delignification was 33%.
Assuntos
Técnicas de Cultura de Células/métodos , Trichoderma/enzimologia , Xilanos/metabolismo , Xilosidases/metabolismo , Xilosidases/isolamento & purificaçãoRESUMO
Xylanase activity was isolated from crude extracts of Trichoderma harzianum strains C and 4 grown at 28oC in a solid medium containing wheat bran as the carbon source. Enzyme activity was demonstrable in the permeate after ultrafiltration of the crude extracts using an Amicon system. The hydrolysis patterns of different xylans and paper pulps by xylanase activity ranged from xylose, xylobiose and xylotriose to higher xylooligosaccharides. A purified ß-xylosidase from the Trichoderma harzianum strain released xylose, xylobiose and xylotriose from seaweed, deacetylated, oat spelt and birchwood xylans. The purified enzyme was not active against acetylated xylan and catalyzed the hydrolysis of xylooligosaccharides, including xylotriose, xylotetraose and xylopentaose. However, the enzyme was not able to degrade xylohexaose. Xylanase pretreatment was effective for hardwood kraft pulp bleaching. Hardwood kraft pulp bleached in the XEOP sequence had its kappa number reduced from 13.2 to 8.9 and a viscosity of 20.45 cp. The efficiency of delignification was 33
Assuntos
Técnicas de Cultura de Células/métodos , Trichoderma/enzimologia , Xilanos/metabolismo , Xilosidases/metabolismo , Xilosidases/isolamento & purificaçãoRESUMO
A low-molecular-weight xylanase activity (XynI) was isolated from the fungus Acrophialophora nainiana after growth in a solid medium containing wheat bran. XynI was purified to apparent homogeneity by ultrafiltration and gel filtration chromatography. The purified enzyme had a molecular weight value of approx. 17 kDa, as determined by SDS-PAGE. This enzyme was most active at 50 degreesC and pH 6.0. At 50 degreesC the half-life was 150 min. The apparent Km value for birchwood xylan was much lower than the Km value for oat spelt xylan. XynI was activated by L-cysteine, DTE, beta-mercaptoethanol, and L-tryptophan. XynI did not show significant sequence homology with other xylanases. The analysis of hydrolysis products of xylans and wood pulps showed that XynI was able to release xylooligomers ranging from X2 to X3 and X2 to X6, respectively. The enzyme was not active against acetylated xylan. A small amount of xylose was released from deacetylated, birchwood, and oat spelt xylans. The results obtained with enzymatic treatment of Kraft pulp indicated a reduction in the amount of chlorine compounds required for the process and enhanced brightness gain.
RESUMO
Two arabinofuranosidases, termed Ara I and Ara II, from solid-state cultures of Penicillium capsulatum were purified to apparent homogeneity as judged by electrophoresis and isoelectric focusing. Each enzyme is a single subunit glycoprotein, and they have M(r)s and pIs of 64,500 and 4.15 (Ara I) and 62,700 and 4.54 (Ara II), respectively. Ara I is most active at pH 4.0 and 60 degrees C, while Ara II exhibits optimal activity at pH 4.0 and 55 degrees C. Ara I is the more thermostable, with its half-life at 70 degrees C and pH 4.0 being 17.5 min. By contrast, the half-life of Ara II is only 9 min at 60 degrees C and pH 4.0. Ara I has the lower Km and higher catalytic constant values with p-nitrophenyl-alpha-L-arabinofuranoside being used as the substrate. Arabinose, a competitive inhibitor (Ki, 16.4 mM) of Ara II, has no effect on Ara I activity at concentrations of up to 40 mM. Each enzyme catalyzes the release of arabinose from pectin, araban, and certain arabinose-containing xylans. The last activity is enhanced by pretreatment of the relevant substrates with xylanase, ferulic acid esterase, or combinations of these enzymes. Thus, arabinoxylooligosaccharides in which arabinose is the sole side chain substituent appear to be the preferred substrates. On the basis of the evidence cited above, each enzyme has been classified as an alpha-L-arabinofuranoside arabinofuranohydrolase (EC 3.2.1.79).