10.542 – Biochemica l Engineerin g
Spring 2005
Surve y o f Biocatalyti c Reactions
Glutam at e Dehydrogenase
Natural: -ketoglu t arate → glutam ate
O
H O OH
H 2 N
H O OH
O O O O
Unnatural: 2-keto-6-hydroxyhexanoic acid → L-6-hydroxynorleucine
O H 2 N
OH OH
H O H O
O O
Leucin e Dehydrogenase
Natural: 2-keto-4-m ethyl-pentanoate → L-leucine
O
H 2 N
OH OH
O O
Unnatural: trim ethylpyruvate → L- ter t -leucine
O
O
O H 2 N
OH OH
Phenylalanin e dehydrogenase
Natural: phenylpyruvate → phenylalanine
O
H 2 N
OH OH
O O
Unnatural: 5-(1,3-dioxola n-2-yl)-2-oxopentanoic acid → 2-am ino-5-(1,3-dioxolan-2- yl)pentanoic acid
O O O H 2 N
OH OH
O O
O O
Lactat e dehydrogenase Natural: pyruvate → lactate
O H O
OH OH
O O
Unnatural: 2-oxo- 4-phenylbutanoic acid → 2-hydroxy-4-phenylbutanoic acid
O
O
OH OH
O OH
Napthalen e dioxygenase
Natural: naphthalene → naphthalene-1,2-diol
OH
OH
Unnatural: 1 H -indole → 1 H -indole-2,3-diol
OH
OH
N N
H H
Lipases
Natural: lipid ↔ glycerol + fatty acids (Note that these enzym e s are also often used in the rev e rse d i rec tion )
O
O R 2
O O
R 1
O
R 3
O
H O OH
H O
O
Ri
3 OH
Unnatural:
(1) 2-oxo-4-phenyla zetidin-3-y l ace tate → 3-hydroxy-4-phenylazetidin-2-one
H N
O
O
H N
OH
O O
(2) 1-pheny lethan am ine + ethyl m e thoxyacetate → ethyl m e thoxyacetate
O
O
NH 2 H N
O
O
O
Subtilisi n (p roteas e)
NH
Natural: peptide bond hydrolysis
NH
R 1
O R 2
O R 3
O O
NH
NH
OH
R 1
R 2 O
H 2 N
NH
O R 3
Unnatural:
F F F
F
O
Me O
N
N
F
O
H
N
N
F
O
Me O
H
N
N
(1)
O
OMe
HO
OM e
+
O O
OM e
D H P M e thy l Es ter
R- D H P Ac i d
S -D HP Me thyl Est e r
(2)
O
O
S
O
O
O
N
O
O
O
S
OH
O
O
N
O
Nitrilase
Natural: acrylonitrile → acrylic acid (acry l ate)
N
OH
O
Unnatural: pyrazine-2-carbonitrile → pyrazine-2-carboxylic acid
N
O
N
N
OH
N N