catalysis/ rothenberg, isbn 978-3-527-31824-7. catalysis: concepts and green applications lecture...
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Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Catalysis: Concepts and Green Applications
Lecture slides for Chapter 5: Biocatalysis
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Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.1
Biocatalysisapplications
Detergents
Food
Fibres
Animal feed
Pulp and paper
Bulk chemicals
Fine chemicals
Pharmaceuticals
Nutraceuticals
Bioremediation
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.2
NO
HOH
OPHO
O
ONH2
O
N
NN
N
NH2
O
OHOH
OPO
OH
Nicotineamide adenine dinucleotide (reduced form, NADH)
O N
NN
N
NH2
O
OHOH
OPO
O
Adenonsine triphosphate (ATP)
O
PO
O
O
PO
O
O N
NN
N
NH2
O
OHO
OPO
O
Coenzyme A (free thiol form, CoASH)
O
PO
O
OPO
O
HSH2CH2CNH
NH
O O
OH
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.3
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.4
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.5
+
+ 25 oC
stable conformer
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.6
NH
HN
O
O
NH3+
NH2
Lys116
Lys115
NH
HN
O
O
NH3+
NH2
Lys116
Lys115
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.7colour fig
N
N
V
OH
OOO
NH
Lys353
His496
N
N
VO
OHOO
NH
Lys353
His496
H2O2
H2O
H2O
HOX
X
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.8
slow
fast
R1 NH
O
R2
Cys25 S
NN
His159
H
R1 NH
O
R2
CysS
NN
His
H
fast
enzyme-bound acyl intermediate
H2O
R1
O
CysS
N:N
His
H2N R2+R1
O
OHH2N R2+
Cys S NN
His
H
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.9
substrate substrate-H2
secondsubstrate
secondsubstrate-H2
NAD(P)H NAD(P)+
enzyme
substrate
secondsubstrate
substrate-H2
secondsubstrate-H2
NAD(P)H NAD(P)+
enzyme I
enzyme II
R
O
R
OH
OH O
R = CH2CH2CH=C(Me)2, n-hexyl, phenyl
OH O
alcohol dehydrogenase
pH = 7.5-8.0
O O
NADPH + H+ NADP+
glucose
gluconic acid
HOOH
OOH
OHOH
OH
enoate reductase
glucose dehydrogenase
pH = 7.5
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.10
O O
O O
+
NH2
NH2
NH
O
OH
HN
OHOenzyme
Eupergit resin
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.11
polymerisation
O NH
HN
O
CONH2
COO
OHN
NH
O=C
NH
O=C
NH
NH2
OHN
HN
O ON,N'-methylenebisacrylamide
+
acryl amide
+
enzyme
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.12
precipitation cross-linking
enzyme molecules~5 nm
aggregates ~1 m
CLEAs,1–100 m
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.13
isatin hydrolase
naphthalene dioxygenase
(NDO)
pathway blocked
tryptophanase
NH
COOH
NH2
NH
NH
OH
OH
cis-indole-2,3-dihydrodiol
NH
OH
indoxyl
[O2]
NH
HN
O
O
indigo
NH
O
indirubin (red pigment)
NHO
tryptophan
indole
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
ampicillin (R = H), amoxycillin (R = OH)
N
S
COOH
HN
O
H
O
NH2
RN
S
COOH
HN
O
H
O
NH2
cephalexine
penicillin acylase
CH3SiCl
PCl5
H2O
n-BuOH
N
S
COOH
HN
O
H
O
N
S
COOH
N
Cl
H
O
N
S
COOH
H2NH
O
6-aminopenicillanic acid (6-APA)
penicillin G
– 40°C
H2O, 37°C
Figure 5.14
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.15
8-aza-1-tetralone
N
O
N N
HN
N
NH2
enzyme
N
NH2
N
NH
EtOAc
+
O
R1 R2
OH
R1 R2
OH+
enzyme
acylatingagent
R1 R2
OAc
R1 R2
OH+
racemisation catalyst
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.16
H2C=O + HCN H2C C N
OH
O-NH4+
O
HOOH
O
HO
NaOH(aq)
glycolonitrile> 99%
nitrilase
ammonium glycolate> 99%
ion-exchange resin
glycolic acid
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.17
P. putida+ O2 + 2H+
OH
OH
O
R O
O
R
O
nradical
polymerisation
O
R O
O
R
O
n
OH
OH
catecholcyclohexa-3,5-diene-1,2-diol
n
+ 2nRCO2H
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.18
amino acid sequence
mutated sequences
cloningselection
screening
select improved sequence
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.19
antigens (haptens)
heavy chain
light chain
antigen binding site
S-S bond hinge
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.20
+ H2OmAb15A10
HO O
+P
O
O
cocaine
ecgonine methyl ester benzoic acidtransition-state mimic
R = peptide anchor group
hydrolysis transition state(approximation)
OMe
O
NMe
OH
OMe
O
NMe
O O
OMe
O
NMe
O OHO
OCH2R
O
NMe
O
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.21
B:B:H
BaseO
OO
BaseO
OHO
PO
O
'5
O-
H:B
'3
BaseO
OO
BaseO
OHO
PO
O
'5
O-
H:B
'3
BaseO
OO
BaseO
OHO
PO
HO
'5
O-
H:B
'3
B:H B:H
B:
BaseO
OHO
PO
'5
O 3'
BaseO
OHO
HO
B:H
BaseO
OHO
PO
'5
O 3'
BaseO
OHOH
HO
B:H
O-O-
BaseO
OHOH
PO
'5
O 3'
BaseO
OHHO
HO
B:HO-
B:H
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.22
D-glucose
pretreatment
reactor series, 55–65 °C
filtration on charcoal
cooling
isoglucose(42% D-fructose)
concentration
HFCS(45% D-fructose)
O
H
HO
H
HO
H
OHOHH H
OHO
OH
OHOH
OH
OHxylose isomerase
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.23
CH2=CHCN + H2O + H2SO4 CH2=CHCONH2.H2SO4
2NH3
CH2=CONH2 + (NH4)2SO4
CH2=CHCN + H2ORaney Cu
80-120 oC96%
selectivity60-80%
conversion
CH2=CHCONH2
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.24
CH2=CHCN + H2O
nitrile hydratase
> 99.9% selectivity
20 oC
> 99.9% conversion
CH2=CHCONH2
CNRR OH
O+ NH3
nitrile hydratase
nitrilase
R NH2
Oamidase
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.25
10-diacetylbaccatin (10-DAB)O
O
O
O
OO
HO
O
O
O
OH
NH
O
OH
O
H
O
O
O
O
OO
HO
HO
HO
OH
H
paclitaxel
11 chemical steps13 reagents13 solvents
plant cell fermentationin aqueous solution
isolation and crystallising
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.26
HOOH
O NH
O O
CO2Me
NH2
benzyloxy-carbonyl(protecting group)
OHO NH
O O
HN
CO2Me
O
OCO2Me
NH2
+
+
CO2Me
NH3 Cl
OHO NH2
HN
CO2Me
O
aspartame (96% yield)
racemisation
DL-methoxyphenylanaline
N-benzyloxycarbonyl-L-aspartic acid
thermolysin
separationMeOH/HCl
hydrogenation
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.27
AlkeneD-glucose + O2
2-keto-D-glucose
Epoxide + H2O
WO
OO
pyranose-2-oxidase
Pd/C D-fructose
H2O2
H2O
H2
AlkeneD-glucose + O2
2-keto-D-glucose
Epoxide + H2O
WO
OO
pyranose-2-oxidase
Pd/C D-fructose
H2O2
H2O
H2
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.28
HO OH
O
pyrophosphatedihydroxyacetone propionaldehyde
OOPO3
2-2-O3P+ +
phosphatase+
aldolase
HO OPO3H2
OO+ OPO3H2
O
OH
OH
OH
O
OH
OH
phosphatase
53% overall yield> 99% ee
+ ...
Catalysis/ Rothenberg, ISBN 978-3-527-31824-7. www.catalysisbook.org
Figure 5.29
dehydration
OH NOCl NO
Cl
NH3
- HCl
NOH
NH2Beckmann
rearrangementNH
O
NH2
,-diaminocaprolactam
(S)-hydrolase
NH2
CO2H
NH2
(S)-lysine
H2O