Abstract
Microbial xylanases that are thermostable, active at alkaline pH and cellulase-free are generally preferred for biobleaching of paper pulp. We screened obligate and facultative marine fungi for xylanase activity with these desirable traits. Several fungal isolates obtained from marine habitats showed alkaline xylanase activity. The crude enzyme from NIOCC isolate 3 (Aspergillus niger), with high xylanase activity, cellulase-free and unique properties containing 580 U l−1 xylanase, could bring about bleaching of sugarcane bagasse pulp by a 60 min treatment at 55°C, resulting in a decrease of ten kappa numbers and a 30% reduction in consumption of chlorine during bleaching. The culture filtrate showed peaks of xylanase activity at pH 3.5 and pH 8.5. When assayed at pH 3.5, optimum activity was detected at 50°C, with a second peak of activity at 90°C. When assayed at pH 8.5, optimum activity was seen at 80°C. The crude enzyme was thermostable at 55°C for at least 4 h and retained about 60% activity. Gel filtration of the 50–80% ammonium sulphate-precipitated fraction of the crude culture filtrate separated into two peaks of xylanase with specific activities of 393 and 2,457 U (mg protein)−1. The two peaks showing xylanase activity had molecular masses of 13 and 18 kDa. Zymogram analysis of xylanase of crude culture filtrate as well as the 50–80% ammonium sulphate-precipitated fraction showed two distinct xylanase activity bands on native PAGE. The crude culture filtrate also showed moderate activities of β-xylosidase and α-l-arabinofuranosidase, which could act synergistically with xylanase in attacking xylan. This is the first report showing the potential application of crude culture filtrate of a marine fungal isolate possessing thermostable, cellulase-free alkaline xylanase activity in biobleaching of paper pulp.
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Acknowledgements
We thank Ms. Gauri Rivonkar and Ms. Shilpa Kamat for the excellent laboratory assistance rendered at N.I.O. Our grateful thanks to Pudumjee Paper Mills Ltd., Pune, India and Seshasayee Paper Mills, Chennai for conducting bleaching trials with our enzyme. This is NIO’s contribution No. 3927.
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Raghukumar, C., Muraleedharan, U., Gaud, V.R. et al. Xylanases of marine fungi of potential use for biobleaching of paper pulp. J IND MICROBIOL BIOTECHNOL 31, 433–441 (2004). https://doi.org/10.1007/s10295-004-0165-2
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DOI: https://doi.org/10.1007/s10295-004-0165-2