Screening and Improving the Recombinant Nitrilases and Application in Biotransformation of Iminodiacetonitrile to Iminodiacetic Acid
作者： Zhi-Qiang Liu , Peter James Baker , Feng Cheng , Ya-Ping Xue , Yu-Guo Zheng
DOI: 10.1371/JOURNAL.PONE.0067197
关键词:
摘要: In this study, several nitrilase genes from phylogenetically distinct organisms were expressed and purified in E. coli order to study their ability mediate the biotransformation of nitriles. We identified three nitrilases: Acidovorax facilis (AcN); Alcaligenes fecalis (AkN); Rhodococcus rhodochrous (RkN), which catalyzed iminodiacetonitrile (IDAN) iminodiacetic acid (IDA). AcN demonstrated 8.8-fold higher activity for IDAN degradation as compared AkN RkN. Based on homology modeling previously described ‘hot spot’ mutations, mutants screened improved activity. One mutant M3 (F168V/L201N/S192F) was identified, demonstrates a 41% enhancement conversion well 2.4-fold catalytic efficiency towards wild-type AcN.
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