Novel xylanases from Simplicillium obclavatum MTCC 9604: comparative analysis of production, purification and characterization of enzyme from submerged and solid state fermentation

Abstract

The production of extracellular xylanase by a newly isolated fungus Simplicillium obclavatum MTCC 9604 was studied in solid-state and submerged fermentation. Multiple xylanases and endoglucanases were produced by the strain during growth on wheat bran in solid state fermentation (SSF). A single xylanase isoform was found to be produced by the same fungus under submerged fermentation (SF) using wheat bran as sole carbon source. Enzyme activity, stability and the protein yield were much higher in SSF than SF. The two dimensional zymogram of the crude enzyme indicated the presence of six isoforms with different pI values starting from pH 3–10. The optimum temperature and pH for the partially purified xylanase activity were 50°C and pH 5.0 respectively; xylanase enzymes exhibited remarkable stability over a broad pH range and the temperature range of 30-60°C which has great potential to be used in biofuels, animal feed and food industry applications.

Isolation, Production, and Characterization of Thermotolerant Xylanase from Solvent TolerantBacillus vallismortisRSPP-15

“…Xylanase was strongly inhibited in the presence of Cu 2+ and Hg 2+ . Similar results were observed in case of Bacillus subtilis [34], Bacillus halodurans PPKS-2 [35], and Simplicillium obclavatum [36]. It has been reported that the xylanase activity was inhibited by Hg 2+ ion, which might be due to its interaction with sulfhydryl groups of cysteine residue in or close to the active site of the enzyme [37].…”

Development of novel economical methodologies for zymographic analysis of purified xylano–pectinolytic enzymes using agrowaste‐based substrates

“…Zymography is used for the detection of enzymatically active protein bands in polyacrylamide gels. It is one of the most effective methodology for studying hydrolytic isozymes …”

“…BCC7928 showed a well‐defined activity band on a gel containing 0.2% wt/vol, Birchwood xylan . Roy et al used Birchwood xylan (2% wt/vol) for zymographic analysis, whereas Cano‐Ramírez et al and Sinma et al reported zymographic analysis of xylanase enzyme using Beachwood xylan substrate (1% wt/vol). Kumar et al supplemented gel with Birchwood xylan for zymographic analysis of xylanase enzyme.…”

“…It is one of the most effective methodology for studying hydrolytic isozymes. [13][14][15] For determining the activity of xylanase and pectinase enzymes, cost-intensive xylan and pectin substrates have been employed by various researchers during zymographic analysis till date. In this study, seven prominent zymographic activity bands (Figure 3a-d Several researchers have used zymographic technique for identifying isozymic forms of xylanase enzymes.…”

Production and applications of xylanases – an overview

“…Studies involving the production of fungal xylanases in solid-state fermentation have shown to obtain very high yields of xylanases as compared with that in the submerged fermentation method. This could be as a result of a poor oxygen supply and shear disruption of fragile fungal biomass in the stirred-tank reactor used for submerged fermentation (Roy et al, 2013;Jain et al, 2013;Gasper et al, 1997).…”