3.5.5.5	A114F	inactive	710940	
3.5.5.5	A116C	the mutant shows activity similar to the wild type enzyme	710940	
3.5.5.5	A116F	the mutant shows about 50% of wild type activity	710940	
3.5.5.5	A136Y	mutant enzyme shows reversed selectivity and preferentially produces (R)-mandelic acid with an enantiomeric excess values of 66.7%	-, 755936	
3.5.5.5	A136Y/I168Y	mutant enzyme produces (R)-mandelic acid with an enantiomeric excess value of 89.7%, an R-enantioselectivity higher than that obtained with the single mutations A136Y and I168Y	755936	
3.5.5.5	A136Y/I168Y/M113G	mutant enzyme with R-selectivity, 90.9% enantiomeric excess	755936	
3.5.5.5	A165E	mutant with very low activities toward (R,S)-mandelonitrile and substrate (R,S)-2-phenylpropionitrile	710930	
3.5.5.5	A165F	decreased degree of amide formation compared to the wild type enzyme and forms about 3% phenylglycine amide from (R,S)-phenylglycinonitrile. In contrast to the wild-type enzyme, the variant almost exclusively forms (R)-phenylglycine. This point mutation results in an almost complete stereoinversion of the reaction	-, 755880	
3.5.5.5	A165F	the mutant enzyme converts racemic mandelonitrile and (R,S)-2-phenylpropionitrile to increased amounts of the R enantiomers of the corresponding acids	710930	
3.5.5.5	A165G	the mutant forms 4.3% amide and thus produces significantly more amide than the wild type enzyme with the substrate (R,S)-2-phenylpropionitrile	710930	
3.5.5.5	A165H	the mutant enzyme shows a significantly increased relative activity for mandelonitrile (compared to (R,S)-2-phenylpropionitrile)	710930	
3.5.5.5	A165R	mutant with very low activities toward (R,S)-mandelonitrile and substrate (R,S)-2-phenylpropionitrile	710930	
3.5.5.5	A165W	the mutant enzyme converts racemic mandelonitrile and (R,S)-2-phenylpropionitrile to increased amounts of the R enantiomers of the corresponding acids	710930	
3.5.5.5	A165Y	the mutant enzyme converts racemic mandelonitrile and (R,S)-2-phenylpropionitrile to increased amounts of the R enantiomers of the corresponding acids	710930	
3.5.5.5	C163A	the mutation results in significantly decreased amounts of amides formed using (R,S)-mandelonitrile and (R,S)-2-phenylpropionitrile as substrates. The mutant demonstrates no significant difference in the enzyme activity compared to the wild type, but shows an extremely low degree of enantioselectivity for the formation of (R)-mandelic acid	710940	
3.5.5.5	C163G	site-directed mutagenesis	-, 719629	
3.5.5.5	C163N	the mutation results in significantly increased amounts of amides formed using (R,S)-mandelonitrile and (R,S)-2-phenylpropionitrile as substrates	710940	
3.5.5.5	C163N/A165R	the mutant demonstrates increased amide formation capacity in comparison to the mutants carrying only single mutations	710940	
3.5.5.5	C163N/W110I	the mutant enzyme forms about 100fold more 2-phenylpropionamide (in relation to the total amount of (R,S)-2-phenylpropionitrile converted) than the wild type, although the relative activity of this mutant for the conversion (R,S)-2-phenylpropionitrile to 2-phenylpropionic acid is only 4% of that observed for the wild type	710940	
3.5.5.5	C163Q	the mutation results in significantly increased amounts of amides formed using (R,S)-mandelonitrile and (R,S)-2-phenylpropionitrile as substrates	710940	
3.5.5.5	C163S	the mutation results in significantly decreased amounts of amides formed using (R,S)-mandelonitrile and (R,S)-2-phenylpropionitrile as substrates. The mutant demonstrates no significant difference in the enzyme activity compared to the wild type, but shows an extremely low degree of enantioselectivity for the formation of (R)-mandelic acid	710940	
3.5.5.5	C164A	the mutant is devoid of nitrilase activity with the substrate (R,S)-2-phenylpropionitrile	710930	
3.5.5.5	E137A	the mutant demonstrates less than 1% of the wild type activity but is still enzymatically competent to convert mandelonitrile to mandelic acid and mandeloamide	-, 733193	
3.5.5.5	E47Q	site-directed mutagenesis	-, 719629	
3.5.5.5	G109F	inactive	710940	
3.5.5.5	H165N	the mutant exhibits a decreased activity for (R,S)-2-phenylpropionitrile compared to the wild type enzyme	-, 734496	
3.5.5.5	H170N	the mutant exhibits a decreased activity for (R,S)-2-phenylpropionitrile compared to the wild type enzyme	-, 734496	
3.5.5.5	I117F	the mutant shows about 50% of wild type activity	710940	
3.5.5.5	I168Y	mutant enzyme shows reversed selectivity and preferentially produces (R)-mandelic acid with an enantiomeric excess values of 74.3%	-, 755936	
3.5.5.5	K129I	site-directed mutagenesis	-, 719629	
3.5.5.5	L111F	the mutant shows about 50% of wild type activity	710940	
3.5.5.5	L113F	the mutant shows activity similar to the wild type enzyme	710940	
3.5.5.5	M113C	mutation enhances the S-enantioselectivity of the enzyme toward mandelonitrile	755936	
3.5.5.5	M113G/A136Y/I168Y	R-selectivity toward mandelonitrile with 90.1% enantiomeric excess	755936	
3.5.5.5	M113L/R128H	S-selectivity toward mandelonitrile with 91.1% enantiomeric excess	755936	
3.5.5.5	M113Q	mutation enhances the S-enantioselectivity of the enzyme toward mandelonitrile	-, 755936	
3.5.5.5	additional information	the C-terminally truncated nitrilase variants demonstrate that deletions up to about 30 amino acids do not significantly change the relevant characteristics of the enzyme, in contrast, longer C-terminal deletions of 47-67 amino acids result in significant decreases in enzyme activities and only about 10% of the activity is found compared with the wild type enzyme, deletion mutants Nit(del-C-20) and Nit(del-C-32) strongly resemble the wild type enzyme, mutants Nit(del-C-47) to Nit(del-C-67) demonstrate reduced nitrilase activities against 2-phenylpropionitrile and show a significant decreased stability of the nitrilase activity compared with the wild type enzyme, deletion mutants also show an increased formation of amide in relation to the acid formation (up to almost 10% compared to 0.2% with the wild type enzyme)	-, 689852	
3.5.5.5	R128E	mutant displays no activity toward mandelonitrile	755936	
3.5.5.5	R128H	mutation enhances the S-enantioselectivity of the enzyme toward mandelonitrile	-, 755936	
3.5.5.5	R128K	mutant demonstrates low enantioselectivity (44.9% enantiomeric excess) as compared with the wild-type enzyme (52.7% enantiomeric excess)	-, 755936	
3.5.5.5	Tyr54C	the mutant shows wild type activity but forms significantly decreased relative amounts of atrolactamide from 2-hydroxy-2-phenylpropionitrile	718648	
3.5.5.5	Tyr54P	the mutant shows wild type activity but forms significantly decreased relative amounts of atrolactamide from 2-hydroxy-2-phenylpropionitrile	718648	
3.5.5.5	Tyr54V	the mutant shows wild type activity but forms significantly decreased relative amounts of atrolactamide from 2-hydroxy-2-phenylpropionitrile	718648	
3.5.5.5	W110F	the mutant shows activity similar to the wild type enzyme	710940	
3.5.5.5	W163A	the mutant exhibits a decreased activity for (R,S)-2-phenylpropionitrile compared to the wild type enzyme	-, 734496	
3.5.5.5	W168A	the mutant forms significantly increases amounts of mandelamide and 2-phenylpropionamide and demonstrates an almost complete inversion of enantioselectivity for the conversion of (R,S)-2-phenylpropionitrile (from R- to S-selectivity)	-, 734496	
3.5.5.5	W188K	the mutant enzyme produces almost exclusively phenylglycine amide from (R,S)-phenylglycinonitrile. In comparison to the wild type, there is an increase in the ee-value of the formed (S)-phenylglycine amide	-, 755880	
3.5.5.5	Y141A	the mutant enzyme is still active and converted the aliphatic (valeronitrile) and the aromatic substrate (2-phenylpropionitrile) with similar relative activities as the wild-type enzyme	-, 758540	
3.5.5.5	Y141W	the mutant enzyme converts valeronitrile with a much higher relative activity than 2-phenylpropionitrile	-, 758540	
3.5.5.5	Y54A	the mutant shows wild type activity but forms significantly decreased relative amounts of atrolactamide from 2-hydroxy-2-phenylpropionitrile	-, 718648	
3.5.5.5	Y54F	the mutant shows wild type activity but forms significantly decreased relative amounts of atrolactamide from 2-hydroxy-2-phenylpropionitrile	-, 718648	
3.5.5.5	Y54H	the mutant shows wild type activity but forms significantly decreased relative amounts of atrolactamide from 2-hydroxy-2-phenylpropionitrile	-, 718648	
3.5.5.5	Y54I	inactive	-, 718648	
3.5.5.5	Y54K	inactive	-, 718648	
3.5.5.5	Y54L	mutant with significantly reduced activity compared to the wild type enzyme	718648	
3.5.5.5	Y54M	the mutant shows wild type activity but forms significantly decreased relative amounts of atrolactamide from 2-hydroxy-2-phenylpropionitrile	718648	
3.5.5.5	Y54R	inactive	718648