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High level expression of a recombinant xylanase by Pichia pastoris NC38 in a 5 L fermenter and its efficiency in biobleaching of bagasse pulp
Authors:Birijlall Natasha  Manimaran Ayyachamy  Kumar Kuttanpillai Santhosh  Permaul Kugen  Singh Suren
Affiliation:Department of Biotechnology and Food Technology, Faculty of Applied Sciences, Durban University of Technology, P.O. Box 1334, Durban 4000, South Africa.
Abstract:A genetically modified XynA gene from Thermomyces lanuginosus was expressed in Pichia pastoris under the control of GAP promoter. P. pastoris expressed greater levels of xylanase (160 IU ml(-1)) on BMGY medium without zeocin after 56 h. The xylanase production by recombinant P. pastoris was scaled up in a 5L fermenter containing 1% glycerol and the highest xylanase production of 139 IU ml(-1) was observed after 72 h. Further studies carried out in fermenter under controlled pH (5.5) yielded a maximum xylanase production of 177 IU ml(-1) after 72 h. The biobleaching efficacy of crude xylanase was also evaluated on bagasse pulp and a brightness of 47.4% was observed with 50 IU of crude xylanase used per gram of pulp, which was 2.1 points higher in brightness than the untreated samples. Reducing sugars (24.8 mg g(-1)) and UV absorbing lignin-derived compounds values were considerably higher with xylanase treated samples.
Keywords:Pichia pastoris   Xylanase   Glycerol aldehyde 3-phosphate promoter (GAP)   Bagasse pulp   Biobleaching
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