Costa L, Brissos V, Lemos F, Ribeiro F Ramôa, Cabral J M S
IBB-Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Instituto Superior Técnico, Lisbon, Portugal.
Bioprocess Biosyst Eng. 2009 Jan;32(1):53-61. doi: 10.1007/s00449-008-0220-x. Epub 2008 Apr 29.
The hydrolysis reaction of p-nitrophenyl butyrate catalyzed by lipases was followed with in situ UV/vis diode array spectrophotometry. Five enzymes - Candida antarctica lipase B and Fusarium solani pisi cutinase wild-type and three single-mutation variants - were tested as catalysts in homogeneous conditions and immobilized on zeolite NaY, on a polyacrylate support and as cross-linked aggregates. Using deconvolution techniques and kinetic modeling, the thermal stability of the different biocatalysts was compared in operational conditions and the results were supported by steady-state enzyme fluorescence measurements. We concluded that both the mutagenesis and the immobilization on zeolite NaY had a positive effect on the thermal stability of F. solani pisi cutinase.
采用原位紫外/可见二极管阵列分光光度法跟踪脂肪酶催化对硝基苯基丁酸酯的水解反应。测试了五种酶——南极假丝酵母脂肪酶B、茄病镰刀菌角质酶野生型及其三个单突变变体——作为均相条件下的催化剂，并将其固定在NaY沸石、聚丙烯酸酯载体上以及制成交联聚集体。运用去卷积技术和动力学模型，在操作条件下比较了不同生物催化剂的热稳定性，稳态酶荧光测量结果支持了这些结果。我们得出结论，诱变和固定在NaY沸石上均对茄病镰刀菌角质酶的热稳定性有积极影响。
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