The application of nitrilases as important biocatalysts in nitrile degradation has attracted considerable attention. With its advanced protein engineering platform established over the years, Creative BioMart has successfully developed engineered nitrilases with catalytic activity, substrate tolerance, and thermal stability. Our scientists are committed to providing fully customized engineered nitrilases production solutions for industry to customers around the world.

Introduction of Nitrilases Production

Nitrilases can hydrolyze nitriles into corresponding carboxylic acids and ammonia, and are ideal for replacing traditional chemical catalysts in fine chemical production. In recent years, nitrilase-mediated biocatalysis has attracted extensive attention from scholars and entrepreneurs. Nitrilases are derived from bacteria, filamentous fungi, yeast and plants. Among them, the industrial production of niacin and (R)-(-)-mandelic acid using nitrilase has demonstrated the great economic potential of nitrilase. To meet the development needs of green industry, there is an urgent need to improve the catalytic performance of existing nitrilase enzymes or to discover the potential of new enzymes. With the advent of new technologies and protocols in biology and computer science, an increasing number of novel nitrilase enzymes have been discovered, purified, and characterized. Protein engineering techniques have been successfully applied to improve the activity, stability and selectivity of nitrilases. Studies have shown that these nitrilases have shown biodegradability and are effective methods for environmental bioremediation for degrading nitrile contaminants generated during fine chemical and pharmaceutical production.

Fig 1. Putative nitrilase active site. (Gong J S, et al., 2012)

Solutions

Due to their excellent selectivity and wide application, nitrilases play an important role in promoting the green development of industry. Creative BioMart has successfully utilized a variety of protein engineering techniques to produce engineered nitrilases with catalytic activity, substrate tolerance, and thermostability for industry. Our customers have widely used nitrilases for carboxylic acid production, bioremediation and surface modification of polymers. In order to provide customized high-quality nitrilases to global customers, we take the following solutions:

• Screening and discovering new nitrilase-producing strains or new nitrile invertases with greater potential.
• Broadening the substrate spectrum of nitrilases, and degrade a variety of nitrile substrates into nitrilases of various carboxylic acids.
• Improving the operational stability of nitrilase. Stable wild-type strains are isolated by certain screening methods, and then the target strains are regenerated by genetic modification to make them more stable.
• Improving the catalytic efficiency of nitrilase. Catalytic efficiency is improved by increasing the solubility of nitrilase substrates in organic solvents.

Applications of Protein Engineering in Nitrilases Production

Protein engineering allows researchers to modify the substrate spectrum of nitrilases, broaden the pH and temperature ranges, and increase the activity and selectivity of nitrilases to meet standards for industrial applications. We have achieved protein engineering modification of nitrilase, significantly improved the catalytic performance of nitrilase, and broadened the scope of industrial application of nitrilase.

• Screening methods for nitrilases

Based on nitrile lyase activity, substrate specificity, and the biochemical properties of the substrate or product, our protein engineers have successfully designed high-throughput screening methods to discover and identify nitrile lyases from natural sources and metagenomic libraries.

• Directed evolution

Our protein engineers use error-prone PCR to incorporate beneficial mutations of recombinantly-generated nitrile lyase variants, increasing their activity and their adaptability at low pH.

• Site-directed mutagenesis

Our protein engineers have modified and improved nitrile lyases using site-directed mutagenesis, resulting in significantly improved biochemical properties.

Creative BioMart has in-depth knowledge and experience of the tools and processes involved in engineered nitrilases development, and provides professional nitrilases production solutions for industry. Whether your nitrilases are in the discovery and screening stages, or are planning engineered nitrilases production, please contact us to discuss further details to ensure your next success.

Reference

1. Gong J S, Lu Z M, Li H, et al.. (2012) Nitrilases in nitrile biocatalysis: recent progress and forthcoming research[J]. Microbial cell factories. 11(1): 1-18.

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