Highly efficient conversion of 1-cyanocycloalkaneacetonitrile using a “super nitrilase mutant”
- 57 Downloads
Nitrilase is the member of carbon–nitrogen hydrogen hydrolase superfamily, which has been widely used for the hydrolysis of nitriles into corresponding carboxylic acids. But most nitrilases are plagued by product inhibition in the industrial application. In this study, a “super nitrilase mutant” of nitrilase with high activity, thermostability and improved product tolerance from Acidovorax facilis ZJB09122 was characterized. Then, an efficient process was developed by employing the whole cell of recombinant E. coli for the conversion of high concentration of 1-cyanocyclohexylacetonitrile-to-1-cyanocyclohexaneacetic acid, an important intermediate of gabapentin. Under the optimized conditions, the higher substrate concentrations such as 1.3 M, 1.5 M and 1.8 M could be hydrolyzed by 13.58 g DCW/L with outstanding productivity (> 740 g/L/day). This study developed a highly efficient bioprocess for the preparation of 1-cyanocyclohexaneacetic acid which has the great potential for industrial application.
KeywordsNitrilase Product tolerance Biocatalysis 1-Cyanocyclohexaneacetic acid Gabapentin
This work was funded by the National Natural Science Foundation of China (nos. 21476210 and 21706235).
- 6.Brady D, Dube N, Petersen R (2006) Green chemistry: highly selective biocatalytic hydrolysis of nitrile compounds. S Afr J Sci 102(7–8):339–344Google Scholar
- 15.Burns MP, Wong JW (2004) Biocatalytic preparation of 1-cyanocyclohexaneacetic acid. WO2004111256 A1. https://patents.google.com/patent/WO2004111256A1
- 20.Zheng RC, Zheng YG, Zhang Q, Huang YM, Li Y, Weng J, Liu T, Fan W (2015) Nitrilase from Arabis alpina, its encoding gene, vector, recombinant bacterial strain and uses thereof. US20170355976 A1. https://patents.google.com/patent/US20170355976A1/
- 21.Kobayashi M, Nagasawa T, Yamada H (1988) Regiospecific hydrolysis of dinitrile compounds by nitrilase from Rhodococcus rhodochrous J1. Appl Microbiol Biotechnol 29(2–3):231–233Google Scholar
- 29.Xue YP, Shu XR, Zou SP, Wang YJ, Zheng YG (2015) Efficient recovery of 1-cyanocyclohexaneacetic acid by ion-exchange process. Sep Sci Technol 50(17):2717–2725Google Scholar