Available online at www.sciencedirect.com
Biocatalysis — key to sustainable industrial chemistryRoland Wohlgemuth
The ongoing trends to process improvements, cost reductions
and increasing quality, safety, health and environment
requirements of industrial chemical transformations have
strengthened the translation of global biocatalysis research
work into industrial applications. One focus has been on
biocatalytic single-step reactions with one or two substrates,
the identification of bottlenecks and molecular as well as
engineering approaches to overcome these bottlenecks.
Robust industrial procedures have been established along
classes of biocatalytic single-step reactions. Multi-step
reactions and multi-component reactions (MCRs) enable a
bottom-up approach with biocatalytic reactions working
together in one compartment and recations hindering each
other within different compartments or steps. The
understanding of the catalytic functions of known and new
enzymes is key for the development of new sustainable
chemical transformations.
Address
Sigma–Aldrich, Industriestrasse 25, CH-9470 Buchs, Switzerland
Corresponding author: Wohlgemuth, Roland
Current Opinion in Biotechnology 2010, 21:713–724
This review comes from a themed issue on
Chemical biotechnology
Edited by Phil Holliger and Karl Erich Jaeger
Available online 26th October 2010
0958-1669/$ – see front matter
# 2010 Elsevier Ltd. All rights reserved.
DOI 10.1016/j.copbio.2010.09.016
IntroductionThe creation of value-added products by chemical trans-
formations has contributed significantly to the quality of
life over the centuries and has reached a high level, but it
has been suggested that many of the stoichiometric
reactions in current use should be replaced by catalytic
processes [1]. Although catalytic tools are not only a
cornerstone of our present economy and society, but also
a key feature of basic life processes, most of the catalysts
used in the automotive, fuel refining, and chemical
industries consist either of inorganic, organometallic or
of organic catalysts in heterogeneous form, as for
example, catalysts involved in pollutant removal from
the exhaust leaving the car engines. The use of biocata-
lysts in chemical transformations has really taken off with
the focus on safe, healthy, resource efficient and econ-
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omical, energy saving, and environment-friendly pro-
duction procedures. The global needs for clean
manufacturing technologies, nonrenewable raw materials,
management of hazardous chemicals and waste present
new research challenges to both chemistry and biotech-
nology. These sciences are taking up these challenges
and the initiatives in Green/sustainable chemistry [2,3]
and white/industrial biotechnology [4] have emerged in
their disciplines independently. It is therefore of crucial
importance for the success of implementation and trans-
lation of science and technology into standard industrial
practice to develop a common chemistry–biotechnology
interface. One common opportunity for improvement and
invention is the current use of protecting groups for
overcoming nonselective and incompatible reactivities
in synthesis and biomimetic as well as enzyme-catalyzed
synthesis can provide the selectivities needed to over-
come barriers [5]. The manufacturing of molecular com-
plexity from simple starting materials with a minimum
number of steps, avoiding protection–deprotection loops
and orientation towards function of the product attract
much interest and biocatalytic process steps are well
positioned for contributing to the solutions of the
above-mentioned challenges [6].
The creation of sustainable value by viable industrial
processes and synthetic pathways requires not only
research progress in chemistry and biotechnology, but
in addition the integration of research from molecular and
engineering sciences, thereby enabling a large range of
industrial biotransformations [7–10]. As reaction devel-
opment serves different practical needs, progress in the
working areas single-step reactions, multi-step reactions,
and multi-component reactions (MCRs) will be discussed
in the following sections. Despite the enormous achieve-
ments in the chemical synthesis of organic compounds,
once believed to be accessible only by biological pro-
cesses and ‘vital forces’, over the past two centuries, many
present state-of-the-art processes are highly inefficient
[3]. This and additional boundary conditions like safety,
health and environment issues in industrial processes
have revitalized the interest in the discovery/invention
of novel biocatalytic reactions and reaction method-
ologies, which have been evolved by nature to achieve
highly efficient and selective transformations. Therefore
the section on the development of new biocatalytic
reaction methodology addresses this important industrial
innovation area.
Industrial biocatalytic single-step reactionsThe early success of single biocatalytic reaction steps
in classical organic synthesis schemes has led to an
Current Opinion in Biotechnology 2010, 21:713–724
714 Chemical biotechnology
increasing number of established industrial processes and
continues to be a useful approach for the introduction of
biocatalysis into industrial practice. The discovery and
development of novel biocatalytic reaction steps can
thereby focus on overcoming synthetic bottleneck reac-
tions and improving the performance of existing chemical
reactions according to industrial requirements. Biocata-
lytic versions of reactions which are impossible or imprac-
tical by existing chemistry tools generate high interest
and stimulate further process research and development
work in industry.
Oxidation and reduction reactions
Oxidations and reductions catalyzed by oxidoreductases
have progressed towards the tools of choice (Figure 1) due
to their improved performance with respect to reaction
selectivity, safety, health, and environment aspects. Se-
lective introduction of one or two oxygen atoms by
biocatalysts has continued to attract a lot of industrial
interest. Among the reactions introducing one oxygen
atom, selective asymmetric hydroxylations, epoxidations,
and Baeyer–Villiger oxidations [11–13,14�] have made
significant progress and are of interest for the oxyfunc-
tionalization of inexpensive organic building blocks. Se-
lective biocatalytic oxidations of one out of several
hydroxygroups, as for example, in alcohols and sugars,
continue to be of industrial interest since the thirties of
the last century and have additional sustainability
benefits compared to the classical chemical oxidations
[7]. Since classical chemical oxidations often use stoichio-
metric oxidants in excess, the selective removal of
remaining oxidants is decisive for the product quality
and enzymatic methods have become standard practice in
production.
Depending on the enzyme properties and the cofactor
recycling system, both the oxidative and the reductive
directions of an oxidoreductase application are of interest
[15,16]. Sustainable enzymatic reductions of aldehydes
and ketones are reliable, scalable and inexpensive routes
to optically active alcohols and have been extensively
employed in organic synthesis despite the vast number of
asymmetric reductions [17]. Even in the area of the
reduction of carbon–carbon double bonds, where catalytic
hydrogenation with hydrogen gas in autoclaves is per-
formed routinely, new asymmetric biocatalytic reductions
of activated alkenes bearing an electron-withdrawing
group have become interesting methods for preparing
the corresponding saturated products in up to >99% ee
and for side stepping the use of hydrogen gas [18]. High
enantioselectivity was also observed for the asymmetric
reduction of activated a,b-unsaturated enones catalyzed
by pentaerythritol tetranitrate reductase for reaction pro-
duct stereogenic centers at the beta-carbon atom [19].
Enoate reductases have also been used for the conversion
of a series of a,b-unsaturated nitriles to the optically
Current Opinion in Biotechnology 2010, 21:713–724
active nitrile products in high yields and excellent enan-
tioselectivities [20].
Amination reactions
As in nature, industrial biocatalytic aminations have been
performed by the two different routes of transamination
and reductive amination. The use of amino acid dehydro-
genases in reductive amination of prochiral precursors
continues to play an important role in the enzymatic
production of D-enantiomers and L-enantiomers of both
natural and non-natural amino acids. Transaminases have
obtained increased interest for the asymmetric synthesis
of amines from prochiral ketones [21–24] and amination is
becoming a key reaction (Figure 2) in industrial biotrans-
formations [9]. New routes to nonchiral amines are also of
interest and a new biocatalytic transamination of pyri-
doxal-50-phosphate has been achieved with complete
conversion [25]. The efficiency of the manufacturing
process for the antidiabetic compound sitagliptin has
been greatly improved by replacing the Rh(Jobiphos)-
catalyzed asymmetric hydrogenation of an enamine at
high pressure with a direct transaminase-catalyzed amin-
ation of prositagliptin ketone [26��]. The best engineered
enzyme could convert 200 g/l prositagliptin ketone to
sitagliptin with an excellent ee of >99.95%. A 53%
productivity increase, 19% waste reduction, elimination
of heavy metals, cost reductions and avoiding specialized
high-pressure hydrogenation equipment have been found
as specific advantages of the biocatalytic process [26��].
Glycosylation reactions
As selective chemical glycosylation reactions require a
substantial synthetic effort involving various protecting
group chemistries in organic solvents, the use of glyco-
syltransferases for coupling glycosyl donors to nonpro-
tected acceptors in aqueous media (Figure 3) continues to
attract a lot of interest [27–29]. Methods based on the
application of glycosyltransferases are currently recog-
nized as being the most effective for the preparation of
complex and highly pure oligosaccharides [30�]. The
trihexosylceramides Gb3 and iGb3 have been synthes-
ized by specific galactosyltransferases using lactosylcer-
amide as acceptor [31]. Sialyltransferases have been used
in chemoenzymatic or whole-cell approaches for the
synthesis of a large library of sialoside standards and
derivatives [32�]. Carbohydrate-based drug design makes
use of various glycosyltransferases for the production of
novel glycosylated compounds, as no single universal
glycosyltransferase has been found [33]. The final hexose
to be transferred from the NDP-hexose to the aglycon can
thereby be diversified by a variety of enzymes like
dehydratases, epimerases and aminotransferases.
Hydrolysis and reverse hydrolysis reactions
On the basis of the vast number of established enzymatic
reactions using hydrolases in aqueous and nonaqueous
systems, this area has become well established and new
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Sustainable industrial chemistry Wohlgemuth 715
Figure 1
Selected biocatalytic oxidation and reduction reactions. The cyclohexanone-monooxygenase-catalyzed Baeyer–Villiger oxidation of bicycloheptenone
has been applied industrially by Sigma–Aldrich with the substrate-feed-product-recovery-technology (SFPR) using Optipore L-493 as adsorber for
high space-time yield [14�].
applications appearing in various fields of organic chem-
istry can build on this experience (Figure 4). The large-
scale availability of many hydrolases like acylases,
amidases, esterases, lipases, proteases and their ease of
use without any cofactors has been a key factor for the
rapid growth of this reaction class in industry [34]. The
robustness and scalability of these reactions with stan-
dard equipment have been useful for resolutions, dera-
cemizations, desymmetrizations in early steps or mild
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deprotections in late steps of a synthesis. The complete
conversion of a substrate into one product of high enan-
tiomeric purity is particularly attractive, as for example,
in desymmetrizations of prochiral diols or diesters. Inex-
pensive acyl donors like acids or simple esters are pre-
ferred for cost-sensitive productions, but require tools to
drive reactions to completion. Lipase-catalyzed acyla-
tions with activated acyl donors like enol esters and acid
anhydrides are practically irreversible. Lipase-catalyzed
Current Opinion in Biotechnology 2010, 21:713–724
716 Chemical biotechnology
Figure 2
Selected biocatalytic amination reactions. The transaminase-catalyzed asymmetric amination of prositagliptin ketone to sitagliptin has been applied
industrially by Merck. Abbreviations: PLP = pyridoxal-50-phosphate; MBA = (S)-a-methylbenzylamine.
polymerization in an organic solvent or one bulk mono-
mer is advantageous in reducing energy consumption and
in polymerizing multifunctional monomers or monomers
which undergo side reactions or are degraded under
process conditions [35]. The enzymatic resolution of a
substrate with a remote stereogenic center has been
realized in the first enantioselective synthesis of (S)-
monastrol [36]. An interesting high yield synthesis of
12-aminolauric acid from v-laurolactam has been devel-
oped by enzymatic transcrystallization using v-laurolac-
tam hydrolase from Acidivorax sp. [37�]. This method has
been chosen because of low conversion ratios by the use
of organic solvents and biphasic systems. Enzymatic
Current Opinion in Biotechnology 2010, 21:713–724
transcrystallization starts with the addition of crystalline
substrate to the aqueous reaction medium, which dis-
solves the substrate up to its solubility limit, and the
enzymatic reaction can then give the soluble product,
which will crystallize, when the product concentration
from the enzymatic conversion exceeds the product
solubility. Overall, the process resembles a SFPR system
[11], where the crystalline substrate is converted into
crystalline product in a highly efficient and environment-
friendly process without organic solvent, acid or alkali. A
nitrilase-catalyzed kinetic resolution of 2-cyano-1,4-ben-
zodioxane and 2-cyano-6-formyl-1,4-benzodioxane to
optically active 1,4-benzodioxane-2-carboxylic acids
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Sustainable industrial chemistry Wohlgemuth 717
Figure 3
Selected biocatalytic glycosylation reactions representing the enormous potential of glycosyltransferases for future industrial applications.
enables mild and enantioselective nitrile hydrolysis
without damage to labile functional groups like the
formyl group [38].
Carbon–carbon formation reactions and carbon–carbon
bond cleaving reactions
The formation and cleavage of carbon–carbon bonds is of
prime importance for constructing the carbon skeleton
not only in synthetic organic chemistry, but also in the
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metabolic pathways of living cells. Among the great
variety of enzymes, hydroxynitrile lyases, aldolases,
and transketolases have attracted much interest
(Figure 5). Hydroxynitrile lyases have been valuable
for manufacturing enantiopure target cyanohydrins from
aldehydes, as versatile bifunctional building blocks for
chemical synthesis [39]. Strategies for overcoming reac-
tion limitations and suppression of nonenzymatic side
reactions combine approaches from enzyme and reaction
Current Opinion in Biotechnology 2010, 21:713–724
718 Chemical biotechnology
Figure 4
Selected biocatalytic hydrolysis and reverse hydrolysis reactions. A recombinant novel isoform of pig liver esterase termed alternative pig liver esterase
(APLE) has been applied industrially by DSM.
engineering [40]. Crude hydroxynitrile lyase has also
been used for the enantioselective cyanohydrin synthesis
in a microreactor [41]. Biocatalysis by means of aldolases
offers a unique stereoselective and green tool to perform
carbon–carbon bond formation or cleavage. Recent
advances in aldolase-catalyzed stereoselective carbon–carbon bond formation reactions are valuable for gener-
ating molecular diversity and for synthetic improvements
from small chiral polyfunctional molecules to highly
Current Opinion in Biotechnology 2010, 21:713–724
complex oligosaccharide analogs [42]. Aldolase-cata-
lyzed carbon–carbon bond formation has been used for
the large-scale synthesis of a chloromethyl-substituted,
a,b-unsaturated d-lactone [43��]. The synthetic poten-
tial of thiamin diphosphate-dependent enzymes for
asymmetric carboligations, such as asymmetric cross-
benzoin condensations, has been extended appreciably
and a variety of enantiomerically pure 2-hydroxyketones
have been synthesized by enzymatic carbon–carbon
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Sustainable industrial chemistry Wohlgemuth 719
Figure 5
Selected biocatalytic carbon–carbon bond formation reactions. The double aldol condensation of acetaldehyde with chloroacetaldehyde catalyzed by
deoxyribose-5-phosphate aldolase (DERA-aldolase) has been applied industrially by DSM in the production of chiral lactones.
bond ligation of aldehydes [44]. The use of benzaldehyde
lyase and benzoylformate decarboxylase in recombinant
Escherichia coli resting cells in a MTBE/aqueous buffer
biphasic medium has improved substrate solubility and
extractive workup [45]. Another route to enantiomeri-
cally pure 2-hydroxy-ketones is the enzymatic chain
elongation of aldehydes by a two-carbon unit, which
can be catalyzed by transketolase and driven to com-
pletion by the use of the irreversible C2-ketol donor b-
hydroxypyruvate [46–48].
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A novel biocatalytic carbon–carbon bond formation reac-
tion equivalent to Friedel–Crafts alkylation has been
catalyzed by methyltransferases using S-adenosyl-L-
methionine and analogs [49��]. A very broad range of
acceptor substrates including cyclic and open-chain
ketones as well as diketones and a-ketoesters and
b-ketoesters have been found in the first enzymatic
asymmetric intermolecular aldehyde–ketone cross-
coupling reaction, using the thiamine-dependent enzyme
YerE [50��]. Changing the substrate specificity of the
Current Opinion in Biotechnology 2010, 21:713–724
720 Chemical biotechnology
Figure 6
Scheme of a biocatalytic multi-step and a biocatalytic multi-component reaction.
carbon–carbon bond forming enzyme tyrosine phenol
lyase has been key for replacing the cumbersome chemi-
cal multi-step synthesis of nonnatural 3-substituted tyro-
sine derivatives by a single-step biocatalytic synthesis
with complete conversion and excellent enantioselectiv-
ity, starting from the corresponding phenol precursor,
pyruvate and ammonia [51��].
Industrial biocatalytic multi-step reactionsMulti-step processes coupling two or more biocatalytic
reactions in one pot (Figure 6) are attractive because of
the reduction in the number of process steps, productivity
improvements and overcoming thermodynamic barriers
[66]. One-pot synthetic methods involving multiple bond
formation steps such as domino, tandem or cascade reac-
tions eliminate also time-consuming recovery and purifi-
cation steps. Biocatalytic reactions have thereby been
combined with other chemical or biocatalytic reactions.
A prominent two-step example is the conversion of cepha-
Current Opinion in Biotechnology 2010, 21:713–724
losporin C to 7-aminocephalosporanic acid by D-amino acid
oxidase and cephalosporin acylase. The synthesis of ator-
vastatin has been achieved by the biocatalytic reduction of
ethyl-4-chloroacetoacetate using a ketoreductase-cata-
lyzed reaction as the first step and a halohydrin-dehalo-
genase-catalyzed substitution reaction of the chloro-
substituent with the cyano-group [52]. Another two-step
reaction sequence has been used in the conversion of an
aromatic alkene to a chiral 2-hydroxy ketone. The carbon–carbon double bond in the olefin trans-anethole to
para-anisaldehyde has been cleaved biocatalytically with
a Trametes hirsuta extract and with molecular oxygen
as oxidant. The second reaction step catalyzed the
condensation of para-anisaldehyde to acetaldehyde by
the enantiocomplementary C–C bond forming enzymes
benzaldehydelyase and benzoylformatedecarboxylase,
respectively, to yield either (R)-2-hydroxy-1-(4-methoxy-
phenyl)-propanone or (S)-2-hydroxy-1-(4-methoxyphe-
nyl)-propanone [53].
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Sustainable industrial chemistry Wohlgemuth 721
Natural product synthesis and modification by biocataly-
tic multi-step reactions is of much interest because of the
challenges in large-scale production of bioactive small
molecules from natural sources or by total synthesis.
Many opportunities exist for preparing a wide range of
natural product variants due to the substrate flexibility of
the pathway enzymes. The bottom-up assembly of plant
biosynthetic pathways in microorganisms is of interest for
exploring the fascinating capabilities of the individual
enzymes as well as for facilitating scalable production
platforms for the synthesis of natural and unnatural alka-
loids [54]. The optimization of biocatalytic pathways to
macrotetrolides is highly attractive, because chemical
synthesis of compounds like nonactin has not been com-
petitive for large-scale production [55]. This is related to
the exquisite selectivity and orthogonality of biocatalytic
functional group transformations, which enables the
organization of multi-step reactions in defined reaction
spaces in an analogous way as in biological cells, cell
compartments or multi-enzyme machineries. The
achievements of biocatalytic multi-step reactions serve
as gold standard for the reaction development in organic
chemistry [4].
Industrial biocatalytic multi-componentreactionsWhile the tactics of step-by-step reactions is based on a
cascade of subsequent functional group transformations,
the goal for MCRs is to construct several bonds between
the components by a parallel operation of different reac-
tions with completely independent reactivity and selec-
tivity. MCRs are therefore step-efficient procedures
converging towards the product and avoiding protecting
group chemistry. MCRs enable building molecular com-
plexity directly from more than two components.
Although MCRs like the Strecker reaction are important
industrial reactions in organic chemistry, the develop-
ment of biocatalytic MCRs (Figure 6) has only recently
attracted interest.
A novel lipase-catalyzed direct Mannich reaction in water
has been discovered, involving aniline, a nonenolizable
substituted benzaldehyde as electrophile and the enoliz-
able acetone as a source of nucleophile [56]. A sequence
of a biocatalytic desymmetrization of a 3,4-substituted
meso-pyrrolidin with monoamine oxidase N from Asper-gillus niger and the use of the resulting enantiopure
1-pyrrolin as component in an Ugi-type 3-component
reaction has been performed in two separate operations
in order to achieve the best yields, diastereomeric ratio
and ee values [57��]. An interesting approach towards a
biocatalytic asymmetric Strecker reaction has combined
transimination with imine-cyanation in a double dynamic
covalent system under thermodynamic control and sub-
sequently coupled in a one-pot process with lipase-
catalyzed transacylation under kinetic control [58]. A
biocatalytic Biginelli 3-component reaction has been
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developed for the high-yield synthesis of 3,4-dihydropyr-
imidine-2-(1H)-ones, consisting of the condensation of
urea or thiourea with a substituted benzaldehyde and a
1,3-ketoester at room temperature in aqueous phosphate
buffer pH 7.0 and using Saccharomyces cerevisiae as bioca-
talyst [59].
Development of new biocatalytic reactionmethodologyIndustrial applications aim at stable processes with
robust, simple and sustainable operation and product
recovery as well as high molecular economy [60–63].
The search for new biocatalytic reaction methodology
is experiencing a boost by progress in a number of
relevant areas like the connection between the broad
set of natural product biosynthetic reactions and the
genes that encode them [64] or the tremendous progress
in engineering enzymes by directed evolution [65��].
Whether the reaction is performed on a small or large
scale, the confinement or localization of enzymes in a
certain reaction space, while retaining their catalytic
activities under process conditions, is key. Expanding
the organic chemistry of enzyme-catalyzed reactions
and interfacing the enzyme reactions with classical chemi-
cal reactions in this reaction space, with no need for using
protecting groups, is promising [66]. A concise approach to
the synthesis of all 24 hexoses and 5-deoxy-hexoses, still
ongoing, is based on a range of biocatalysts which inter-
convert polyols and ketoses, aldose isomerases for the
equilibration of ketoses and aldoses, and D-tagatose-3-
epimerase for the C-3 equilibration of a wide range of
substrates like ketoses, deoxysugars, and C-branched
sugars [67�]. An interesting biocatalytic domino reaction
between phenol and various cyclic 1,3-dicarbonyl com-
pounds yielded annulated benzofurans, using the
enzymes tyrosinase and laccase from Agaricus bisporus [68].
The capturing and activation of carbon dioxide by
enzymes has obtained increased interest [69], as on the
one hand the chemistry of direct carboxylation reactions
is underdeveloped and on the other hand many carbox-
ylating and decarboxylating enzymes are occurring widely
in nature. The novel continuous flow enzymatic carbox-
ylation of pyrrole to pyrrole-2-carboxylate by immobilized
Bacillus megaterum represents an interesting green engin-
eering approach [70]. Salicylic acid decarboxylase from
Trichosporon monilliforme has been discovered to catalyze
the enzymatic Kolbe–Schmitt reaction from phenol to
salicylic acid [71]. 3,4-Dihydroxybenzoate decarboxylase
from Enterobacter cloacae enabled the mild regioselective
carboxylation of catechol to 3,4-dihydroxy-benzoic acid
with 3 M potassium hydrogencarbonate at 308C [72].
ConclusionsBiocatalytic single-step reaction platforms developed over
the last years have progressed rapidly in the industrial
Current Opinion in Biotechnology 2010, 21:713–724
722 Chemical biotechnology
production environment and many more methodologies
developed at the research scale are waiting to be applied
and to be scaled up. Discovery and development of novel
biocatalytic single-step reactions continues to be import-
ant, especially in areas where no direct functional group
transformation is known or where the known chemical
transformation is lacking safety, selectivity or sustainabil-
ity. The innate selectivity and orthogonality advantage of
biocatalytic reactions bears a lot of potential for major
improvements in multi-step reactions. Attention needs
to be paid to both the molecular and the engineering
aspects of the architecture of such biocatalytic multi-step
systems. Whatever route is selected, key to further
advances in sustainable chemical reactions is the devel-
opment of novel biocatalytic reaction methodologies,
which are modular, scalable, and compatible with the
development of chemical reactions. The science, technol-
ogy and industry of chemical synthesis and catalysis on the
other hand is accepting established biocatalytic reaction
platforms, because of the need for method and route
simplification, molecular economy, safety, health, and
environment improvements. Therefore the knowledge
building in industrial biocatalysis and its practical imple-
mentation is key for value creation in a future bioeconomy.
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24. Ward J, Wohlgemuth R: High-yield biocatalytic aminationreactions in organic synthesis. Curr Org Chem 2010,14:1914-1927.
25. Schell U, Wohlgemuth R, Ward JM: Synthesis of pyridoxamine50-phosphate using an MBA: pyruvate transaminase asbiocatalyst. J Mol Catal B: Enzym 2009, 59:279-285.
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Savile CK, Janey JM, Mundorff EC, Morre JC, Tam S, Jarvis WR,Colbeck JC, Krebber A, Fleitz FJ, Brands J et al.: Biocatalyticasymmetric synthesis of chiral amines from ketones appliedto sitagliptin manufacture. Science 2010, 329:305-309.
A variety of enzyme engineering techniques have been applied to thecreation of a transaminase biocatalyst with the required properties andactivity toward the prositagliptin ketone. This has resulted in an efficientbiocatalytic trans-amination process to replace a rhodium-catalyzedasymmetric hydrogenation for the large-scale manufacturing of the anti-diabetic compound sitagliptin.
27. Chokhawala HA, Huang S, Lau K, Yu H, Cheng J, Thon V, Hurtado-Ziola N, Guerrero JA, Varki A, Chen X: Combinatorialchemoenzymatic synthesis and high-throughput screening ofsialosides. ACS Chem Biol 2008, 3:567-576.
28. Wohlgemuth R: Tools and ingredients for the biocatalyticsynthesis of carbohydrates and glycoconjugates. BiocatalBiotransformation 2008, 26:42-48.
29. Wanga Z, Gilbert M, Eguchi H, Yu H, Cheng J, Muthanad S, Zhou L,WangPG, Chen X, Huang X: Chemoenzymatic syntheses oftumor-associated carbohydrate antigen Globo-H and
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stage-specific embryonic antigen 4. Adv Synth Catal 2008,350:1717-1728.
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Weijers CAGM, Franssen MCR, Visser GM: Glycosyltransferase-catalyzed synthesis of bioactive oligosaccharides. BiotechnolAdv 2008, 26:436-456.
Review on the effective application of glycosyltransferases for the pre-paration of complex and highly pure oligosaccharides.
31. Adlercreutz D, Weadge JT, Petersen BO, Duus JØ, Dovichi NJ,Palcic MM: Enzymatic synthesis of Gb3 and iGb3 ceramides.Carbohydr Res 2010, 345:384-388.
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Chen X, Varki A: Advances in the biology and chemistry of sialicacids. ACS Chem Biol 2010, 5:163-176.
A sialic acid review including recent advances in chemoenzymatic synth-esis as well as large-scale E. coli.
33. Luzhetskyy A, Mendez C, Salas JA, Bechthold A:Glycosyltransferases, important tools for drug design. CurrTop Med Chem 2008, 8:680-709.
34. Wohlgemuth R: Large-scale applications of hydrolasesin biocatalytic asymmetric synthesis. In Large-scaleAsymmetric Catalysis. Edited by Blaser HU, Federsel HJ.Weinheim: Wiley-VCH; 2010.
35. Gross RA, Ganesh M, Lu W: Enzyme-catalysis breathes new lifeinto polyester condensation polymerizations. TrendsBiotechnol 2010, 28:435-443.
36. Blasco MA, Thumann S, Wittmann J, Giannis: A, Groger H:Enantioselective biocatalytic synthesis of (S)-monastrol.Bioorg Med Chem Lett 2010, 20:4679-4682.
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Fukuta Y, Komeda H, Yoshida Y, Asano Y: High yield synthesis of12-aminolauric acid by ‘enzymatic transcrystallization’ of v-laurolactam using v-laurolactam hydrolase from Acidivoraxsp. T31. Biosci Biotechnol Biochem 2009, 73:980-986.
An interesting high-yield enzymatic hydrolysis of v-laurolactam by v-laurolactam hydrolase from Acidivorax sp. has been developed. Crystal-line v-laurolactam, added to the enzyme solution, has been converted tocrystalline 12-aminolauric acid with the high volume yield of >200 g/l,high purity and >97% conversion.
38. Benz P, Muntwyler R, Wohlgemuth R: Chemoenzymaticsynthesis of chiral carboxylic acids via nitriles. J Chem TechnolBiotechnol 2007, 82:1087-1098.
39. Purkarthofer T, Skranc W, Schuster C, Griengl H: Potential andcapabilities of hydroxynitrile lyases as biocatalysts in thechemical industry. Appl Microbiol Biotechnol 2007, 76:309-320.
40. Andexer JN, Langermann JV, Kragl U, Pohl M: How to overcomelimitations in biotechnological processes—examples fromhydroxynitrile lyase applications. Trends Biotechnol 2009,27:599-607.
41. Koch K, van den Berg RJF, Nieuwland PJ, Wijtmans R,Schoemaker HE, van Hest JCM, Rutjes FPJT: Enzymaticenantioselective C–C-bond formation in microreactors.Biotechnol Bioeng 2008, 99:1028-1033.
42. Clapes P, Fessner WD, Sprenger GA, Samland AK: Recentprogress in stereoselective synthesis with aldolases. Curr OpinChem Biol 2010, 14:154-167.
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Wolberg M, Dassen BHN, Schurmann M, Jennewein S,Wubbolts MG, Schoemaker HE, Mink D: Large-scale synthesisof new pyranoid building blocks based on aldolase-catalysedcarbon–carbon bond formation. Adv Synth Catal 2008,350:1751-1759.
Large-scale aldolase-catalyzed carbon–carbon bond formation, basedon a reaction discovered by CH Wong and coworkers, has permitted thehighly stereoselective synthesis of substituted d-lactones.
44. Muller M, Gocke D, Pohl M: Thiamin diphosphate in biologicalchemistry: exploitation of diverse thiamin diphosphate-dependent enzymes for asymmetric chemoenzymaticsynthesis. FEBS J 2009, 276:2894-2904.
45. Dominguez e Maria P, Stillger T, Pohl M, Kiesel M, Liese A,Groger H, Trauthwein H: Enantioselective C–C bond ligationusing recombinant Escherichia coli-whole-cell biocatalysts.Adv Synth Catal 2008, 350:165-173.
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46. Wohlgemuth R: C2-ketol elongation by transketolase-catalyzedasymmetric synthesis. J Mol Catal B: Enzym 2009,61:23-29.
47. Shaeri J, Wright I, Rathbone EB, Wohlgemuth R, Woodley JM:Characterization of enzymatic D-xylulose 5-phosphatesynthesis. Biotechnol Bioeng 2008, 101:761-767.
48. Wohlgemuth R, Smith MEB, Dalby PA, Woodley JM:Transketolases. In Encyclopedia of Industrial Biotechnology.Edited by Flickinger MC. Hoboken, NJ: Wiley; 2010.
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Stecher H, Tengg M, Ueberbacher BJ, Remler P, Schwab H,Griengl H, Gruber-Khadjawi M: Biocatalytic Friedel–Craftsalkylation using non-natural cofactors. Angew Chem Int Ed2009, 48:9546-9548.
The SAM-dependent methyltransferases NovO from Streptomyces spher-oides and CouO from Streptomyces rishiriensis, cloned and expressed inE. coli, have been shown to accept modified cofactors and to catalyze thesynthesis of a range of monosubstituted methylated, allylated, propargy-lated and benzylated arenes with excellent regioselectivity.
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Lehwald P, Richter M, Rohr C, Liu HW, Muller M: Enantioselectiveintermolecular aldehyde–ketone cross-coupling through anenzymatic carboligation reaction. Angew Chem Int Ed 2010,49:1-5.
The thiamindiphosphate-dependent enzyme YerE has been shown tocatalyze the asymmetric cross-coupling of aldehydes and ketones tochiral teriary alcohols.
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Seisser B, Zinkl R, Gruber K, Kaufmann F, Hafner A, Kroutil W:Cutting long syntheses short: access to non-natural tyrosinederivatives employing an engineered tyrosine phenol lyase.Adv Synth Catal 2010, 352:731-736.
The laborious and time-consuming multi-step synthesis of 3-substitutedtyrosine derivatives has been replaced by a single biocatalytic one-stepreaction using an engineered tyrosine phenol lyase.
52. Ma SM, Gruber J, Davis C, Newman L, Gray D, Wang A, Grate J,Huisman GW, Sheldon RA: A green-by-design biocatalyticprocess for atorvastatin intermediate. Green Chem 2010,12:81-86.
53. Kurlemann N, Lara M, Pohl M, Kroutil W, Liese A: Asymmetricsynthesis of chiral 2-hydroxy ketones by coupled biocatalyticalkene oxidation and C–C bond formation. J Mol Catal B:Enzymatic 2009, 61:111-116.
54. Leonard E, Runguphan W, O’Connor S, Jones Prather K:Opportunities in metabolic engineering to facilitate scalablealkaloid production. Nat Chem Biol 2009, 5:292-300.
55. Jani P, Emmert J, Wohlgemuth R: Process analysis ofmacrotetrolide biosynthesis during fermentation by means ofdirect infusion LC–MS. Biotechnol J 2008, 3:1-7.
56. Li K, He T, Li C, Feng XW, Wang N, Yu XQ: Lipase-catalyzeddirect Mannich reaction in water: utilization of biocatalyticpromiscuity for C–C bond formation in a ‘one-pot’ synthesis.Green Chem 2009, 11:777-779.
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Znabet A, Ruijter E, Decanter FJJ, Kohler V, Helliwell M, Turner NJ,Orru RVA: Highly stereoselective synthesis of substitutedprolyl peptides using a combination of biocatalyticdesymmetrization and multicomponent reactions. AngewChem Int Ed 2010, 49:5289-5292.
A highly diastereoselective Ugi-multicomponent reaction of opticallyactive 3,4-disubstituted 1-pyrrolines, obtained by monoamineoxidaseN-catalyzed desymmetrization of the corresponding meso-pyrrolidines,with isocyanides and carboxylic acids has been developed for thesynthesis of substituted prolylpeptides.
58. Pornrapee V, Ramstrom O: Dynamic asymmetricmulticomponent resolution: lipase-mediated amidation of adouble dynamic covalent system. J Am Chem Soc 2009,131:14419-14425.
59. Kumar A, Maurya RA: An efficient baker’s yeast catalyzedsynthesis of 3,4-dihydropyrimidin-2-(1H)-ones. TetrahedronLett 2007, 48:4569-4571.
60. Wohlgemuth R: Modular and scalable biocatalytic tools forpractical safety, health and environmental improvements inthe production of speciality chemicals. BiocatalBiotransformation 2007, 25:178-185.
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61. Wohlgemuth R: Tools and ingredients for the biocatalyticsynthesis of metabolites. Biotechnol J 2009, 9:1253-1265.
62. Tao J, Xu JH: Biocatalysis in development of greenpharmaceutical processes. Curr Opin Chem Biol 2009, 13:43-50.
63. Wohlgemuth R: Green production of fine chemicals by isolatedenzymes. In Biocatalysis for Green Chemistry and ChemicalProcess Development. Edited by Tao JA, Kazlauskas RJ.Hoboken, NJ: Wiley; 2010.
64. Walsh CT, Fischbach MA: Natural products version 2.0:connecting genes to molecules. J Am Chem Soc 2010,132:2469-2493.
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Reetz MT: Directed evolution of enantioselective enzymes: anunconventional approach to asymmetric catalysis in organicchemistry. J Org Chem 2009, 74:5767-5778.
An excellent perspective on the principles, strategies and methods of thedirected evolution of enantioselective enzymes and their successes andfuture challenges in their applications as asymmetric catalysts in organicchemistry.
66. Wohlgemuth R: Interfacing biocatalysis and organic synthesis.J Chem Technol Biotechnol 2007, 82:1055-1062.
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Rao D, Best D, Yoshihara A, Gullapalli P, Morimoto K,Woemaid MR, Wilson FX, Izumori K, Fleet GWJ: A conciseapproach to the synthesis of all twelve 5-deoxyhexoses:D-tagatose-3-epimerase — a reagent that is both specific andgeneral. Tetrahedron Lett 2009, 50:3559-3563.
Current Opinion in Biotechnology 2010, 21:713–724
An interesting equilibration of 5-deoxy-D-fructose to 5-deoxy-D-psicoseand of 5-deoxy-L-psicose to 5-deoxy-L-fructose, providing substrates forthe preparation of all D-5-deoxy-aldohexoses and L-5-deoxy-aldo-hexoses.
68. Leutbecher H, Hajdok S, Braunberger C, Neumann M, Mika S,Conrad J, Beifuss U: Combined action of enzymes: the firstdomino reaction catalyzed by Agaricus bisporus. Green Chem2009, 11:676-679.
69. Glueck SM, Gumus S, Fabian WMF, Faber K: Biocatalyticcarboxylation. Chem Soc Rev 2010, 39:313-328.
70. Matsuda T, Marukado R, Koguchi S, Nagasawa T, Mukouyama M,Harada T, Nakamura K: Novel continuous carboxylation usingpressurized carbon dioxide by immobilized decarboxylase.Tetrahedron Lett 2008, 49:6019-6020.
71. Yoshida T, Inami Y, Matsui T, Nagasawa T: Regioselectivecarboxylation of catechol by 3,4-dihydroxybenzoatedecarboxylase of Enterobacter cloacae P. Biotechnol Lett 2010,32:701-705.
72. Kirimura K, Gunji H, Wakayama R, Hattori T, Ishii Y: EnzymaticKolbe–Schmitt reaction to form salicylic acid from phenol:enzymatic characterization and gene identification of a novelenzyme, trichosporon moniliiforme salicylic aciddecarboxylase. Biochem Biophys Res Commun 2010, 394:279-284.
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