腈化合物生物转化反应的调控及在高选择性合成中的应用

The productivity of nitrile hydratase(NHase) and amidase from Rhodococcus sp. AJ270 was investigated. The influence of pH, temperature, genetic mutation (UV mutation), immobilization conditions on the activity of NHase and amidase was systematically studied. The NHase and amidase genes from AJ270 were amplified by PCR and the recombinant plasmid pBNH and pBAM were constructed and expressed in E. coli., and the NHase and amidase gene sequences of AJ270 were determined. In addition, amidase from AJ270 was purified through (NH4)2SO4, DEAE-Sphacel, Phenyl Sepharose and Sephadex G-200. This work provided a basis to obtain the microbial mutant and genetic engineering strains of high activity and selectivity via biotechnology and gene engineering in genetic modification of nitrilases gene.

项目旨在运用生物技术和基因工程方法对腈解酶类的基因进行遗传改造，研究获得具有活性和高选择性微生物突变体和基因工程菌的有效方法，为腈化合物的生物转化提供新的生物催化剂，以实现腈化合物的高选择性物转化反应的调控，专一性的合成一些重要羧酸及其衍生物。.........