Klatte Stephanie, Wendisch Volker F
Chair of Genetics of Prokaryotes, Faculty of Biology & CeBiTec, Bielefeld University, Universitaetsstr. 25, 33615 Bielefeld, Germany.
Chair of Genetics of Prokaryotes, Faculty of Biology & CeBiTec, Bielefeld University, Universitaetsstr. 25, 33615 Bielefeld, Germany.
Bioorg Med Chem. 2014 Oct 15;22(20):5578-85. doi: 10.1016/j.bmc.2014.05.012. Epub 2014 May 17.
Whole cell biotransformation is an upcoming tool to replace common chemical routes for functionalization and modification of desired molecules. In the approach presented here the production of various non-natural (di)amines was realized using the designed whole cell biocatalyst Escherichia coli W3110/pTrc99A-ald-adh-ta with plasmid-borne overexpression of genes for an l-alanine dehydrogenase, an alcohol dehydrogenase and a transaminase. Cascading alcohol oxidation with l-alanine dependent transamination and l-alanine dehydrogenase allowed for redox self-sufficient conversion of alcohols to the corresponding amines. The supplementation of the corresponding (di)alcohol precursors as well as amino group donor l-alanine and ammonium chloride were sufficient for amination and redox cofactor recycling in a resting buffer system. The addition of the transaminase cofactor pyridoxal-phosphate and the alcohol dehydrogenase cofactor NAD(+) was not necessary to obtain complete conversion. Secondary and cyclic alcohols, for example, 2-hexanol and cyclohexanol were not aminated. However, efficient redox self-sufficient amination of aliphatic and aromatic (di)alcohols in vivo was achieved with 1-hexanol, 1,10-decanediol and benzylalcohol being aminated best.
全细胞生物转化是一种即将出现的工具，可替代用于所需分子功能化和修饰的常见化学途径。在此提出的方法中，使用设计的全细胞生物催化剂大肠杆菌W3110/pTrc99A-ald-adh-ta实现了各种非天然（二）胺的生产，该催化剂通过质粒介导过表达用于L-丙氨酸脱氢酶、醇脱氢酶和转氨酶的基因。通过L-丙氨酸依赖性转氨作用和L-丙氨酸脱氢酶的级联醇氧化反应，实现了醇向相应胺的氧化还原自足转化。在静止缓冲系统中，添加相应的（二）醇前体以及氨基供体L-丙氨酸和氯化铵足以进行胺化反应和氧化还原辅因子循环。无需添加转氨酶辅因子磷酸吡哆醛和醇脱氢酶辅因子NAD(+)即可实现完全转化。仲醇和环醇，例如2-己醇和环己醇未发生胺化反应。然而，通过体内对脂肪族和芳香族（二）醇进行有效的氧化还原自足胺化反应，实现了对1-己醇、1,10-癸二醇和苯甲醇的最佳胺化。
