Shoaib Azhar

Extraction of hemicelluloses from fiberized spruce wood

A novel mechanical pre-treatment method was used to separate the wood chips into fiber bundles in order to extract high molecular weight wood polymers. The mechanical pre-treatment involved chip compression in a conical plug-screw followed by defibration in a fiberizer. The fiberized wood was treated with hot water at various combinations of time and temperature in order to analyze the extraction yield of hemicelluloses at different conditions. Nearly 6 mg/g wood of galactoglucomannan was obtained at 90 °C/120 min which was about three times more than what could be extracted from wood chips. The extracted carbohydrates had molecular weight ranging up to 60 kDa. About 10% of each of the extracted material had a molecular weight above 30 kDa. The extraction liquor could also be reused for consecutive extractions with successive increase in the extraction yield of hemicelluloses.

Stabilization of Polysaccharides During Alkaline Pre-Treatment of Wood Combined with Enzyme-Supported Extractions in a Biorefinery

Specific enzymes have demonstrated an ability to increase the possibilities for extracting wood polymers. Enzymatic treatment requires an open wood structure, which was achieved by extended impregnation of the wood. However, lignin and some of the hemicelluloses, primarily glucomannan, were lost during the impregnation. To improve the carbohydrate yield, three glucomannan modification agents—sodium borohydride, polysulphide, and anthraquinone—were used, which increased the yields of the impregnated materials from 76.6% to 89.6%, 80.0%, and 81.3%, respectively. Through the use of additives, most of the glucomannan could be retained in the wood while still allowing the enzymes to penetrate the wood and attack the polymers. The additives also increased the extraction yield from 9 to 12% w/w wood. Gamanase treatment prior to the extraction increased the extraction yield to 14%. Of the three stabilizing agents, sodium borohydride was the most efficient, providing the highest extraction yields.

Biochemical characterization of the novel endo-β-mannanase AtMan5-2 from Arabidopsis thaliana

Plant mannanases are enzymes that carry out fundamentally important functions in cell wall metabolism during plant growth and development by digesting manno-polysaccharides. In this work, the Arabidopsis mannanase 5-2 (AtMan5-2) from a previously uncharacterized subclade of glycoside hydrolase family 5 subfamily 7 (GH5_7) has been heterologously produced in Pichia pastoris. Purified recombinant AtMan5-2 is a glycosylated protein with an apparent molecular mass of 50kDa, a pH optimum of 5.5-6.0 and a temperature optimum of 25°C. The enzyme exhibits high substrate affinity and catalytic efficiency on mannan substrates with main chains containing both glucose and mannose units such as konjac glucomannan and spruce galactoglucomannan. Product analysis of manno-oligosaccharide hydrolysis shows that AtMan5-2 requires at least six substrate-binding subsites. No transglycosylation activity for the recombinant enzyme was detected in the present study. Our results demonstrate diversification of catalytic function among members in the Arabidopsis GH5_7 subfamily.