Xylanase production by Trichoderma harzianum rifai by solid state fermentation on sugarcane bagasse

Abstract

Sugarcane bagasse was used as substrate for xylanase production by means of a strain of Trichoderma harzianum Rifai isolated from decaying Aspidosperma sp. (peroba) wood. The bagasse was washed, dried, milled and wetted with minimal salts medium and the cultures grown at 28 ± 2ºC for 7 days. Two extraction methods were tested for enzyme recovery: (A) Tween 80, 0.1% (v/v), in physiological saline, and (B) 50mM sodium acetate buffer, pH 5.0, under agitation (180rpm) for 15, 30 and 60min. After a single extraction, both extraction methods recovered an average of 15U/ml of xylanase activity, independent on the time of shaking. A second and third extraction recovered 10.4 and 6.6U/ml xylanase, respectively. The effect of volume size for extraction, and sugarcane bagasse concentration, on xylanase production were also investigated. The growth profile of Trichoderma harzianum was followed over 20 days on 14% (w/v) bagasse, and highest xylanase activity (288U/ml) appeared on the seventh day. The enzymatic extract after precipitation with ammonium sulphate was submitted to electrophoresis on polyacrylamide gels and showed 4 protein-staining bands, one of which exhibited xylanase activity.

Obtaining New Cultures Of Microorganisms That Produces Cellulases And Xylanases From The Sugarcane Bagasse

“…Both zymograms using CMC and xylan as substrate showed a signal around 20 kDa (Figure 2, lanes 3 and 5). The weaker signal in the xylan zymogram indicates the presence of a~20 kDa xylanolytic activity which is compatible with other studies of this species that describe xylanase activities around this molecular mass with the lack of cellulase activity (Rezende et al 2002;Lee et al 2009;do Vale et al 2012). The L04 cellulase activity around 20 kDa could correspond to the endoglucanase (EGIII) from T. harzianum IOC3844 characterized by Generoso et al (2012) described with a low molecular mass, lack the cellulose binding domain (CBD) and able to degrade amorphous cellulose such as CMC.…”

Xylanolytic enzymes production by Aspergillus niger GS1 from solid-state fermentation on corn stover and their effect on ruminal digestibility

“…These results agree with those reported by Rezende et al (2002), and Okafor et al (2007) where xylanase activity varied from 3.4 to 13 U (mg protein) -1 . These authors used supplemented agricultural by-products such as sawdust, sugarcane pulp, wheat bran, and oat spelt xylan as substrates using either T. harzianum or A. niger ANL 301.…”

On-Site Enzyme Production by Trichoderma asperellum for the Degradation of Duckweed

“…Other factors that improve cellulase production by T. reesei include relative humidity and temperature [28,29], continuous light exposure [27] and aeration and higher substrate concentration (Pandey, 2003) [30]. In a study performed on T. asperellum, a marginally higher endoglucanase and β-xylosidase activity was measured when it was incubated in darkness under a controlled relative humidity of 90%, compared with the standard culture without relative humidity control [10].…”