Preparative scale Baeyer-Villiger biooxidation at high concentration using recombinant Escherichia coli and in situ substrate feeding and product removal process
Preparative scale Baeyer-Villiger biooxidation at high concentration using recombinant Escherichia coli and in situ substrate feeding and product removal process
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DOI：
10.1038/nprot.2007.532
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发表时间：
2008-01-01
期刊：
影响因子：
14.8
通讯作者：
Alphand, Veronique
中科院分区：
文献类型：
--
作者：
Hilker, Iris;Gutierrez, Maria C.;Alphand, Veronique
基于使用吸附树脂（原位底物进料和产物去除）的高效生物催化过程克服了底物和产物抑制造成的限制，使得高底物浓度下的实验成为可能。该工艺已成功应用于 (-)-(1S, 5R)-双环[3.2.0]hept-2-en-6-one (25 g) 的制备规模Baeyer-Villiger 生物氧化。使用过表达环己酮单加氧酶的重组大肠杆菌（1升）的全细胞作为生物催化剂，并将底物预加载到吸附树脂上。得到相应的内酯，产率为75-80%。细胞生长和生物转化的时间各约为24小时，可以通过使用定制的气泡柱来改善氧气供应。
An efficient biocatalytic process based on the use of adsorbent resin (in situ substrate feeding and product removal) makes experiments at high substrate concentration possible by overcoming limitations due to substrate and product inhibition. This process was successfully applied to the preparative scale Baeyer-Villiger biooxidation of (-)-(1S, 5R)-bicyclo[3.2.0]hept-2-en-6-one (25 g). Whole cells of recombinant E. coli (1 liter) overexpressing cyclohexanone monooxygenase were used as a biocatalyst and the substrate was preloaded onto the adsorbent resin. The corresponding lactone was obtained in 75-80% yield. Time for cell growth and biotransformation is about 24 h each and oxygen supply can be improved by using a tailor-made bubble column.