1.4.3.12: cyclohexylamine oxidase

kcat/Km for 1-(4-methoxybenzyl)-1,2,3,4,5,6,7,8-octahydroisoquinoline is 30% of the wild-type value

F327A

kcat/Km for 1-(4-methoxybenzyl)-1,2,3,4,5,6,7,8-octahydroisoquinoline is 179% of the wild-type value

G202A

kcat/Km for 1-(4-methoxybenzyl)-1,2,3,4,5,6,7,8-octahydroisoquinoline is 298% of the wild-type value

I173A

kcat/Km for 1-(4-methoxybenzyl)-1,2,3,4,5,6,7,8-octahydroisoquinoline is 67% of the wild-type value

I201A

kcat/Km for 1-(4-methoxybenzyl)-1,2,3,4,5,6,7,8-octahydroisoquinoline is 24% of the wild-type value

L200A

kcat/Km for 1-(4-methoxybenzyl)-1,2,3,4,5,6,7,8-octahydroisoquinoline is 150% of the wild-type value

L295A

kcat/Km for 1-(4-methoxybenzyl)-1,2,3,4,5,6,7,8-octahydroisoquinoline is 30% of the wild-type value

L295I

kcat/Km for 1-(4-methoxybenzyl)-1,2,3,4,5,6,7,8-octahydroisoquinoline is 73% of the wild-type value

P396A

kcat/Km for 1-(4-methoxybenzyl)-1,2,3,4,5,6,7,8-octahydroisoquinoline is 1177% of the wild-type value

T203A

kcat/Km for 1-(4-methoxybenzyl)-1,2,3,4,5,6,7,8-octahydroisoquinoline is 63% of the wild-type value

F317A

kcat/Km for 1-(4-methoxybenzyl)-1,2,3,4,5,6,7,8-octahydroisoquinoline is 30% of the wild-type value

G202A

kcat/Km for 1-(4-methoxybenzyl)-1,2,3,4,5,6,7,8-octahydroisoquinoline is 298% of the wild-type value

I201A

kcat/Km for 1-(4-methoxybenzyl)-1,2,3,4,5,6,7,8-octahydroisoquinoline is 24% of the wild-type value

L200A

kcat/Km for 1-(4-methoxybenzyl)-1,2,3,4,5,6,7,8-octahydroisoquinoline is 150% of the wild-type value

T203A

kcat/Km for 1-(4-methoxybenzyl)-1,2,3,4,5,6,7,8-octahydroisoquinoline is 63% of the wild-type value

L199A

the mutant shows generally lower activity (decrease of 15-97%) towards most substrates compared to wild type enzyme with the exception of the larger substrates, such as cyclooctanamine and bicyclic (S)-1-aminotetraline

724095

L199F

L199I

the mutant is more active than the wild type enzyme toward the primary amines

L199T

L353M

the mutant shows 7-445% higher activity towards primary aliphatic amines with cycloalkane or aromatic moieties

724095

M226A

M226F

the mutant shows reduced catalytic efficiency with (S)-1-phenylethanamine and increased catalytic efficiency with 2-methyl-1,2,3,4-tetrahydroquinoline compared to the wild type enzyme

741481

M226I

the mutant is more active than the wild type enzyme toward the primary amines

M226T

T198A

the mutant exhibits enhanced activity relative to the wild type enzyme for most (S)-enantiomers of primary amines and some secondary amines

T198F

T198F/L199S

the mutant exhibits 240times higher catalytic efficiency with 2-methyl-1,2,3,4-tetrahydroquinoline than the wild type enzyme. The mutant shows reduced catalytic efficiency with (S)-1-phenylethanamine compared to the wild type enzyme

T198F/L199S/M226F

T198I

Y321A

Y321F

Y321I

Y321I/ M226T

29.8fold increase of kcat/Km for the substrate L-valine ethyl ester as compared to wild-type enzyme. Chemoenzymatic deracemization is applied to prepare D-valine from racemic valine ethyl ester or L-valine ethyl ester in high yield (up to 95%) with excellent optical purity (more than 99% enantiomeric excess) by employing cyclohexylamine oxidase (CHAO) variant Y321I/M226T exhibiting catalytic efficiency that is 30 times higher than that of the wild type enzyme

Y321I/ M226T/L199F

mutant failed to be expressed

Y321I/L199F

mutant failed to be expressed

Y321I/M226T

the double mutant acts on (S)-N-(prop-2-yn-1-yl)-2,3-dihydro-1H-inden-1-amine

Y321I/M226T/T198I

15.1fold increase of kcat/Km for the substrate L-valine ethyl ester as compared to wild-type enzyme

Y321I/M226T/T198I/F199F

7.5fold increase of kcat/Km for the substrate L-valine ethyl ester as compared to wild-type enzyme

Y321I/T198I

14.2fold increase of kcat/Km for the substrate L-valine ethyl ester as compared to wild-type enzyme

Y321T

the mutation enhances the enzyme activity toward the secondary amines

Y459T

the mutant shows reduced catalytic efficiency with (S)-1-phenylethanamine and increased catalytic efficiency with 2-methyl-1,2,3,4-tetrahydroquinoline compared to the wild type enzyme

741481

L199A

L199F

2.1fold increase of kcat/Km for the substrate L-valine ethyl ester as compared to wild-type enzyme

L199T

the mutant shows reduced catalytic efficiency with (S)-1-phenylethanamine and increased catalytic efficiency with 2-methyl-1,2,3,4-tetrahydroquinoline compared to the wild type enzyme

L353M

the mutant shows 7-445% higher activity towards primary aliphatic amines with cycloalkane or aromatic moieties

M226A

the mutant displays an enhanced activity (5-400%) towards most substrates

M226F

the mutant shows reduced catalytic efficiency with (S)-1-phenylethanamine and increased catalytic efficiency with 2-methyl-1,2,3,4-tetrahydroquinoline compared to the wild type enzyme

M226T

1.3fold increase of kcat/Km for the substrate L-valine ethyl ester as compared to wild-type enzyme

T198F

the mutant shows reduced catalytic efficiency with (S)-1-phenylethanamine and increased catalytic efficiency with 2-methyl-1,2,3,4-tetrahydroquinoline compared to the wild type enzyme

T198F/L199S

T198F/L199S/M226F

the mutant exhibits 406times higher catalytic efficiency with 2-methyl-1,2,3,4-tetrahydroquinoline than the wild type enzyme. The mutant shows reduced catalytic efficiency with (S)-1-phenylethanamine compared to the wild type enzyme

T198I

2.55fold increase of kcat/Km for the substrate L-valine ethyl ester as compared to wild-type enzyme

Y321A

the mutant shows higher catalytic efficiency towards cyclooctanamine compared to the wild type enzyme

Y321F

the mutant shows higher catalytic efficiency towards cyclooctanamine compared to the wild type enzyme

Y321I

13fold increase of kcat/Km for the substrate L-valine ethyl ester as compared to wild-type enzyme

Y321I/T198I