Alcaligenes faecalis nitrilase

Hydrolysis of aliphatic dinitriles into cyano carboxylic acids by nitrilases (nitrilase 1 from Alcaligenes faecalis, nitrilase 2 from Bradyrhizobium Japonicum) [50, 51],... 

Schreiner U, Hecher B, Obrowsky S, Waich K, Klempier N, SteinkeUner G, Gruber K, RozzeU JD, GUeder A, Winkler M. Directed evolution of Alcaligenes faecalis nitrilase [J]. Enzyme Microb Technol 2010,47 (4) 140-146. 

Li S, Yang X. Catalytic properties improvements of Alcaligenes faecalis nitrilase by self-assembly induced aggregation [J]. 2014,31 (Supplement) S44. 

Table 8.8 Relative activity (%) of Alcaligenes faecalis JM3 and Arthrobacter sp. F-73 nitrilases in water-acetone mixtures... 

Figure 8.10 Relative specific activity of Y296 mutations of Alcaligenes faecalis Nitl650 nitrilase for conversion of racemic 2-chloromandelonitrile Y296A (A), Y296F (F), Y296C (C), Y296G (G), Y296N (N), Y296T (T), and Y296S (S)...

Figure 8.11 Relative specific activity of Alcaligenes faecalis Nitl650 nitrilase ( ) and Y296C mutant nitrilase ( ) for conversion of unsubstituted and substituted mandelonitrile mandelonitrile (MN), 2-chloromandelonitrile (2-CMN), 3-chloromandelonitrile (3-CMN), 4-chlormandelonitrile (4-CMN), 4-bromomandelonitrile (4-BMN), and 4-methylmandelonitrile (4-MeMN)...

All nitrilases belonging to aromatic nitrilase and arylacetonitrilase are susceptible to SH-reagents, except the R. rhodochrous K22 aliphatic nitrilase [81]. Analysis by site-directed mutagenesis for our three nitrilases from R. rhodochrous J1 [83], R. rhodochrous K22 [84] and Alcaligenes faecalis JM3 [85] revealed that a unique cysteine that is conserved at the corresponding position in each nitrilase is essential for the catalytic activity (Fig. 7). All nitrilases whose structural genes are cloned have a similar amino acid sequence nitrilase forms a superfamily. 

Nagasawa, T., Manger, 1., and Yamada, H. 1990. A novel nitrilase, arylacetonitrilase, of Alcaligenes faecalis 1M3. Purification and characterization. European Journal of Biochemistry, 194 765-72. 

Yamamoto, K., Fujimatsu, L, and Komatsu, K. 1992. Purification and characterization of the nitrilase from Alcaligenes faecalis ATCC8750 responsible for the enantioselective hydrolysis of mandelonitrile. Journal of Fermentation and Bioengineering, 73 425-30. 

An enantioselective nitrilase has also been shown to be applicable in the dynamic kinetic resolution of mandelonitrile. Using the nitrilase produced by Alcaligenes faecalis ATCC 8750 Yamamoto et al. showed that they could derive (Rj-(-)-mandelic acid from mandelonitrile in 91% yield with an ee of 100%. Under the reaction conditions used non-reacting (S) -mandelonitrile undergoes spontaneous racemiza-tion leading to the high yield (see Scheme 12.1-8)[48]. Currently (R)-mandelic acid and (R)-chloromandelic acid are produced using nitrilases on an industrial scale by the Mitsubishi Rayon Corp. 

Singh R, Banerjee A, Kaul P et al. (2005) Release of an entmtioselective nitrilase from Alcaligenes faecalis MTCC 126 a comparative study. Bioproc Biosys Eng 27(6) 415 24 Singh RS, Rajesh D, Munish P (2007) Partial purification and characterization of exoinulinase from Kluyveromyces marxianus YS-1 or preparation of high-fructose syrup. J Microbiol Biotechnol 178(5) 733-738... 

Fusarium solani [114], Arthrobacter sp. [115], Escherichia coli, transformed with a KLehsiella ozaenae plasmid DNA [116], Rhodococcus rhodochrous J1 [117,118], and Alcaligenes faecalis JM3 [119, 120]. The enzyme of Rhodococcus rhodochrous J1 was employed for the production of p-aminobenzoic acid from p-aminobenzonitrile [121], and nicotinic acid from 3-cyanopyridine [122]. The conversion of 3 anopyridine to nicotinic add, by a nitrilase of Nocardia rhodochrous IjL100-2, was also reported by Vaughan et al. [123]. These nitrilases were usually inactive on aliphatic nitriles. More recently, a new nitrilase, that acts preferentially on aliphatic nitriles, was purified and characterized in Rhodococcus rhodochrous K22 [124]. 

A different method consisting of directed evolution was used with the nitrilases from P. fluorescens [26] and Alcaligenes faecalis [27]. The amide production by the former and the operational stability of the latter at a low pH were improved in this... 

Detzel, C., Maas, R., and Jose, J. (2011). Autodisplay of nitrilase from Alcaligenes faecalis in E. coli yields a whole cell biocatalyst for the S5mthesis of enantiomerically pure (R)-mandelic acid. ChemCatChem, 3, 719-725. 

Synthesis of (8)-mandelic acid via dynamic kinetic resolution of mandelonitrile with a wild-type whole-cell catalyst from Alcaligenes faecalis containing a nitrilase. 

Zhang J-E, Liu Z-Q, Zheng Y-G. Improvement of nitrilase production from a newly isolated Alcaligenes faecalis mutant for biotransformation of iminodiacefonitrile to iminodiacetic acid [J]. 7Taiwan Inst Chem Eng 2012,44 (2) 169-176. 

Figure 6 Synthesis of (J )-mandelic acid from (J ,S)-mandelonitrile by an (R)-specific nitrilase from Alcaligenes faecalis ATCC 8750.

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