Microplate bioassay for determining substrate selectivity of candida rugosa lipase

• Autores: Shi-zhen Wang, Baishan Fang
• Localización: Journal of chemical education, ISSN 0021-9584, Vol. 89, Nº 3, 2012, págs. 409-411
• Idioma: inglés
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• Resumen

  • Substrate selectivity of Candida rugosa lipase was tested using p-nitrophenyl esters of increasing chain length (C1, C7, C15) using the high-throughput screening method. A fast and easy 96-well microplate bioassay was developed to help students learn and practice biotechnological specificity screen. The kinetic constants Vmax and Km of each substrate were compared to indicate the lipase activity and selectivity to different substrates. In this experiment, students were exposed to the core concepts of Michaelis-Menten kinetic analysis and learned the principles and techniques of studying substrate selectivity, including the use of a microplate reader and the associated software, the layout of bioassays, and the multichannel-repeater pipetting.