Nitrilase and Its Application as a ‘Green’ Catalyst

Savan, Ram; Sharma, Rakesh Kumar; Tewari, Neetu; Geetanjali, .; Rawat, Diwan S.

doi:10.1002/cbdv.200690131

Cited by 91 publications

(27 citation statements)

References 87 publications

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“…Nitrilases (EC 3.5.5.1) have attracted substantial interest as a valuable alternative in organic chemical processes, which can catalyze hydrolysis of nitriles to the corresponding carboxylic acids in a single step, and do not produce large amounts of by-products [14]. Especially advantageous is that the enzyme-catalyzed hydrolysis of a variety of nitriles can be highly stereoselective [16].…”

Section: Discussionmentioning

confidence: 99%

Enantioselective biocatalytic hydrolysis of (R,S)-mandelonitrile for production of (R)-(−)-mandelic acid by a newly isolated mutant strain

Xue

Liu

et al. 2010

J Ind Microbiol Biotechnol

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(R)-(-)-Mandelic acid (R-MA) is an important intermediate with broad uses. Recently, R-MA production using nitrilase has been gaining more and more attention due to its higher productivity and enantioselectivity. In this work, a new bacterium WT10, which exhibited favorable nitrilase activity and excellent enantioselectivity for production of R-MA by enantioselective biocatalytic hydrolysis of (R,S)-mandelonitrile, was isolated and identified as a strain of Alcaligenes faecalis. In order to improve its nitrilase activity for industrial application, the wild-type strain WT10 was further subjected to mutagenesis using a combined LiCl-ultraviolet irradiation and low energy N(+) ion beams implantation technique. A valuable mutant strain A. faecalis ZJUTB10 was obtained. The nitrilase specific activity of the mutant strain was greatly improved up to 350.8 U g(-1), in comparison with wild-type strain WT10 of 53.09 U g(-1). The reaction conditions for R-MA production by mutant strain A. faecalis ZJUTB10 were also optimized. Nitrilase activity in mutant strain showed a broad pH optimum at pH 7.7-8.5. The optimal temperature was 35°C. The highest production rate reached 9.3 mmol h(-1) g(-1). The results showed that mutant strain A. faecalis ZJUTB10 was a new candidate for efficient R-MA production from (R,S)-mandelonitrile and could potentially be used in industrial production.

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Section: Discussionmentioning

confidence: 99%

Enantioselective biocatalytic hydrolysis of (R,S)-mandelonitrile for production of (R)-(−)-mandelic acid by a newly isolated mutant strain

Xue

Liu

et al. 2010

J Ind Microbiol Biotechnol

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“…Chemical hydrolysis of nitriles typically requires strongly acidic or basic conditions and higher reaction temperatures, and usually produces unwanted byproducts. In contrast, nitrilasemediated hydrolysis of nitriles to carboxylic acids is of great industrial interest owing to the desirability of conducting such conversions under mild conditions that would not alter other labile reactive groups combined with high yield and selectivity [5,6]. In this regard, nitrilases (EC 3.5.5.1) from a variety of microorganisms and their applications in the production of nicotinic acid, ibuprofen, acrylic acid and the detoxification of cyanide wastes have been reported [7,8].…”

Section: Introductionmentioning

confidence: 93%

Cloning and biochemical properties of a highly thermostable and enantioselective nitrilase from Alcaligenes sp. ECU0401 and its potential for (R)-(−)-mandelic acid production

Zhang

et al. 2010

Bioprocess Biosyst Eng

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A nitrilase gene from Alcaligenes sp. ECU0401 was cloned and overexpressed in Escherichia coli BL21 (DE3) in a soluble form. The encoded protein with a His₆-tag was purified to nearly homogeneity as revealed by SDS-PAGE with a molecular weight of approximately 38.5 kDa, and the holoenzyme was estimated to be composed of 10 subunits of identical size by size exclusion chromatography. The V(max) and K(m) parameters were determined to be 27.9 μmol min⁻¹ mg⁻¹ protein and 21.8 mM, respectively, with mandelonitrile as the substrate. The purified enzyme was highly thermostable with a half life of 155 h at 30 °C and 94 h at 40 °C. Racemic mandelonitrile (50 mM) could be enantioselectively hydrolyzed to (R)-(-)-mandelic acid by the purified nitrilase with an enantiomeric excess of 97%. The extreme stability, high activity and enantioselectivity of this nitrilase provide a solid base for its practical application in the production of (R)-(-)-mandelic acid.

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“…The extensive investigations for the selective transformation of nitriles by biocatalysts over the past decade has revealed an impressive number of enzymes, which possess the desired broad substrate spectrum by retaining their exquisite natural specificities with respect to regioand stereo-selectivity. The ability of enzymes to hydrolyze nitriles, in particular, employed as whole microorganisms, is well established, and a range of both aromatic and aliphatic nitriles as well as dinitriles has been successfully hydrolyzed [9][10][11].…”

Section: Introductionmentioning

confidence: 99%

Optimization of nitrilase production from Alcaligenes faecalis MTCC 10757 (IICT-A3): effect of inducers on substrate specificity

Nageshwar

Sheelu

Shambhu

et al. 2010

Bioprocess Biosyst Eng

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Microbial nitrilases are biocatalysts of interest and the enzyme produced using various inducers exhibits altered substrate specificity, which is of great interest in bioprocess development. The aim of the present study is to investigate the nitrilase-producing Alcaligenes faecalis MTCC 10757 (IICT-A3) for its ability to transform various nitriles in the presence of different inducers after optimization of various parameters for maximum enzyme production and activity. The production of A. faecalis MTCC 10757 (IICT-A3) nitrilase was optimum with glucose (1.0%), acrylonitrile (0.1%) at pH 7.0. The nitrilase activity of A. faecalis MTCC 10757 (IICT-A3) was optimum at 35 °C, pH 8.0 and the enzyme was stable up to 6 h at 50 °C. The nitrilase enzyme produced using different inducers was investigated for substrate specificity. The enzyme hydrolyzed aliphatic, heterocyclic and aromatic nitriles with different substitutions. Acrylonitrile was the most preferred substrate (~40 U) as well as inducer. Benzonitrile was hydrolyzed with almost twofold higher relative activity than acrylonitrile when it was used as an inducer. The versatile nitrilase-producing A. faecalis MTCC 10757 (IICT-A3) exhibits efficient conversion of both aliphatic and aromatic nitriles. The aromatic nitriles, which show not much or no affinity towards nitrilase from A. faecalis, are hydrolyzed effectively with this nitrilase-producing organism. Studies are in progress to exploit this organism for synthesis of industrially important compounds.

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Nitrilase and Its Application as a ‘Green’ Catalyst

Cited by 91 publications

References 87 publications

Enantioselective biocatalytic hydrolysis of (R,S)-mandelonitrile for production of (R)-(−)-mandelic acid by a newly isolated mutant strain

Enantioselective biocatalytic hydrolysis of (R,S)-mandelonitrile for production of (R)-(−)-mandelic acid by a newly isolated mutant strain

Cloning and biochemical properties of a highly thermostable and enantioselective nitrilase from Alcaligenes sp. ECU0401 and its potential for (R)-(−)-mandelic acid production

Optimization of nitrilase production from Alcaligenes faecalis MTCC 10757 (IICT-A3): effect of inducers on substrate specificity

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