Protein engineering of nitrilase for chemoenzymatic production of glycolic acid

Wu, Shijin; Fogiel, Arthur J.; Petrillo, Kelly L.; Jackson, Raymond J.; Parker, Kimberley N.; DiCosimo, Robert; Ben‐Bassat, Arie; O’Keefe, Daniel P.; Payne, Mark S.

doi:10.1002/bit.21643

Cited by 44 publications

(19 citation statements)

References 8 publications

(8 reference statements)

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“…Reacting formaldehyde with carbon monoxide in the presence of an acid catalyst, such as HF, also produces glycolic acid. It can be made using glycolonitrile in an enzymatic process [12,144]. Due to the extremely hazardous nature of reactants involved in these routes, other ways to produce glycolic acid have been investigated.…”

Section: Glycolic Acidmentioning

confidence: 99%

Catalytic Conversion of Lignocellulosic Biomass to Value-Added Organic Acids in Aqueous Media

Lin

Liu

et al. 2014

Green Chemistry and Sustainable Technology

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The transition from today's fossil-based economy to a sustainable economy based on renewable biomass is driven by the concern of climate change and anticipation of dwindling fossil resources. Although biofuels are the central theme of the transition, biomass resources cannot completely replace petroleum. It is projected that biofuels can only supply up to 30 % of today's transportation fuel market even if all available domestic biomass resources are used for the production of liquid fuels. Therefore, transformation of biomass into high-value-added chemicals is advantageous to secure optimal use of the abundant, but limited, biomass resources from the economical and ecological perspective. Industry is increasingly considering bio-based chemical production as an attractive area for investment. The potential for chemical and polymer production from biomass is substantial. The US Department of Energy recently issued a report which listed 12 chemical building blocks considered as potential building blocks for the future. Organic acids (e.g., succinic, lactic, levulinic acid, etc.) are among the widely spread ''platform-molecules,'' which may be further converted into possibly derivable high-value-added chemicals. The transition from a fossil chemical industry to a renewable chemical industry will likewise depend on our ability to focus research and development efforts on the most promising alternatives. In this chapter, we review the emerging technologies on catalytic conversion of biomass to value-added organic acids in aqueous media.

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Section: Glycolic Acidmentioning

confidence: 99%

Catalytic Conversion of Lignocellulosic Biomass to Value-Added Organic Acids in Aqueous Media

Lin

Liu

et al. 2014

Green Chemistry and Sustainable Technology

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“…The first synthetic applications for nitrilases have been established for aromatic nitrilases (especially by using the enzyme from Rhodococcus rhodochrous J1), and examples for the conversion of compounds such as 3-cyanopyridine to nicotinic acid or cyanopyrazine to pyrazinoic acid have been investigated (24,30). Subsequently, several biotransformation reactions which utilize aliphatic nitrilases or arylacetonitrilases were also described (35,52,53). Arylacetonitrilases are able to convert ␣-substituted phenylacetonitriles and are therefore interesting for the enantioselective synthesis of various ␣-substituted carboxylic acids (and amides).…”

Section: Discussionmentioning

confidence: 99%

Conversion of Sterically Demanding α,α-Disubstituted Phenylacetonitriles by the Arylacetonitrilase from Pseudomonas fluorescens EBC191

Baum

Williamson

Sewell

2012

Appl Environ Microbiol

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The nitrilase from Pseudomonas fluorescens EBC191 converted 2-methyl-2-phenylpropionitrile, which contains a quaternary carbon atom in the ␣-position toward the nitrile group, and also similar sterically demanding substrates, such as 2-hydroxy-2-phenylpropionitrile (acetophenone cyanohydrin) or 2-acetyloxy-2-methylphenylacetonitrile. 2-Methyl-2-phenylpropionitrile was hydrolyzed to almost stoichiometric amounts of the corresponding acid. Acetophenone cyanohydrin was transformed to the corresponding acid (atrolactate) and amide (atrolactamide) at a ratio of about 3.4:1. The (R)-acid and the (S)-amide were formed preferentially from acetophenone cyanohydrin. A homology model of the nitrilase suggested that steric hindrance with amino acid residue Tyr54 could impair the binding or conversion of sterically demanding substrates. Therefore, several enzyme variants that carried mutations in the respective residues were generated and subsequently analyzed for the substrate specificity and enantioselectivity of the reactions. Enzyme variants that demonstrated increased relative activities for the conversion of acetophenone cyanohydrin were identified. The chiral analysis of these reactions demonstrated peculiar reaction kinetics, which suggested that the enzyme variants converted the nonpreferred (S)-enantiomer of acetophenone cyanohydrin with a higher reaction rate than that of the (preferred) (R)-enantiomer. Recombinant whole-cell catalysts that simultaneously produced the nitrilase from P. fluorescens EBC191 and a plant-derived (S)-oxynitrilase from cassava (Manihot esculenta) converted acetophenone plus cyanide at pH 4.5 to (S)-atrolactate and (S)-atrolactamide. These recombinant cells are promising catalysts for the synthesis of stable chiral quaternary carbon centers from ketones.

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“…[11] Several other literature reports exist which show the successful application of directed evolution as a tool for protein engineering. [21][22][23][24][25] The general advantage of nitrilases to perform hydrolysis under mild reaction conditions such as room temperature and neutral pH [26] is well known. Therefore it was our goal to generate a new nitrilase variant with enhanced activity for a broad range of substrates.…”

Section: Improved Fitness Of Arabidopsis Thaliana Nitrilasementioning

confidence: 99%