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Faculty of Applied Sciences

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    Evaluation of chitosan–coated magnetic nanoparticle-immobilized thermostable hemicellulases for enhanced saccharification and production of bioethanol
    (2022-09) Mdlaka, Sibongile Patience; Singh, Suren; Puri, Adarsh Kumar
    Enhancing the efficiency of saccharification of pentose and hexose sugars present in lignocellulosic biomass is a major bottleneck for industrial bioethanol production. This problem can be addressed by a concerted effort combining nanotechnology, enzymology and fermentation technology. Functionalized chitosan-coated magnetic nanoparticles (CCMNPs) were prepared and used for co-immobilization of purified xylan hydrolysing xylanase and xylosidase from the thermophilic fungus Thermomyces lanuginosus SSBP for the release of xylose. Stability studies revealed that immobilized enzymes were more stable than free enzymes over a wide range of pH (4.0 – 7.0) and temperature (40 – 90 °C) for xylanase and 30 – 80 °C for xylosidase. The optimum activity of the co-immobilized enzymes shifted slightly as compared to the free enzymes, with coimmobilized xylanase and xylosidase showing optimum activity at pH 6.5 and 6.0, respectively. The study showed sustained production of xylose as the major fermentable sugar under repeated batch and fed-batch saccharification of lignocellulosic biomass. Statistical optimization of saccharification of 1% xylan using response surface methodology indicated the enhanced release of xylose at 50 °C, pH 7.0 and enzyme dose of 60 U/mL xylanase and 30 U/mL xylosidase. Finally, liberated xylose was fermented with Scheffersomyces stipitis to yield bioethanol.
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    Display of phytase on the cell surface of Saccharomyces cerevisiae to degrade phytate phosphorus and improve bioethanol production
    (Springer Verlag, 2016) Xiao, Yan; Shen, Wei; Govender, Algasan; Zhang, Liang; Xianzhong, Chen
    Currently, development of biofuels as an alternative fuel has gained much attention due to resource and environ-mental challenges. Bioethanol is one of most important and dominant biofuels, and production using corn or cassava as raw materials has become a prominent technology. However, phytate contained in the raw material not only decreases the efficiency of ethanol production, but also leads to an increase in the discharge of phosphorus, thus impacting on the environment. In this study, to decrease phytate and its phos-phorus content in an ethanol fermentation process, Saccharomyces cerevisiae was engineered through a surface-displaying system utilizing the C-terminal half of the yeast α-agglutinin protein. The recombinant yeast strain, PHY, was constructed by successfully displaying phytase on the surface of cells, and enzyme activity reached 6.4 U/g wet biomass weight. Ethanol productions using various strains were com-pared, and the results demonstrated that the specific growth rate and average fermentation rate of the PHY strain were higher 20 and 18 %, respectively, compared to the control strain S. cerevisiae CICIMY0086, in a 5-L bioreactor process by simultaneous saccharification and fermentation. More im-portantly, the phytate phosphorus concentration decreased by 89.8 % and free phosphorus concentration increased by 142.9 % in dry vinasse compared to the control in a 5-L bio-reactor. In summary, we constructed a recombinant S. cerevisiae strain displaying phytase on the cell surface, which could improve ethanol production performance and effectively reduce the discharge of phosphorus. The strain reported here represents a useful novel engineering platform for developing an environment-friendly system for bioethanol production from a corn substrate.