Biotechnology Letters 26: 463–472, 2004.

© 2004 Kluwer Academic Publishers. Printed in the Netherlands.

463

Review

Applied biocatalysis for the synthesis of natural flavour compounds –

current industrial processes and future prospects

J. Schrader1,∗ , M.M.W. Etschmann1, D. Sell1 , J.-M. Hilmer2 & J. Rabenhorst2

1 KarlWinnacker Institute, DECHEMA e.V., Theodor-Heuss-Allee 25, 60486 Frankfurt/Main, Germany
2 Symrise GmbH & Co. KG, Mühlenfeldstr. 1, 37603 Holzminden, Germany
∗ Author for correspondence (Fax: +49 69 7564 388; E-mail: schrader@dechema.de)

Received 26 November 2003; Revisions requested 17 December 2003; Revisions received 19 January 2004; Accepted 22 January 2004

Key words: biocatalysis, γ -decalactone, flavour, 2-phenylethanol, vanillin

Abstract
The industrial application of biocatalysis for the production of natural flavour compounds is illustrated by a
discussion of the production of vanillin, γ -decalactone, carboxylic acids, C6 aldehydes and alcohols (‘green
notes’), esters, and 2-phenylethanol. Modern techniques of molecular biology and process engineering, such as
heterologous expression of genes, site-directed mutagenesis, whole-cell biocatalysis in biphasic systems, and
cofactor regeneration for in vitro oxygenation, may result in more biocatalytic processes for the production of
flavour compounds in the future.

Scope of review activities to use biocatalysts for the production of fla-

This review focusses on the industrial application of
biocatalysis for the synthesis of natural flavour com-
pounds. The authors’ intention is to illustrate examples
of bioprocesses currently used or under investiga-
tion in industry rather than to present the tremendous
variety of flavour compounds generally accessible by
biotechnology (for a comprehensive overview see Ber-
ger 1995). The biocatalytic approaches presented have
been compiled to emphasize the fact that quite differ-
ent forms of biocatalysts – from isolated enzymes to
crude plant material to whole microbial cells – and
of bioprocesses – from aqueous to biphasic to organic
systems – can be of industrial importance.
This review is rounded off by a section on recent
advances in genetic and bioprocess engineering which
could lead more biocatalytic approaches to industrial
application in the future.
Introduction
The rising popularity of natural products during the
last decades has triggered off significant research
vour compounds: products derived from bioprocesses
starting with natural substrates are in principle defined
as ‘natural’ if they have already been identified in
plants or other natural sources. Although the con-
sumers’ preference for ‘natural’, ‘bio’ or ‘organic’
products is undoubtedly an important market pull in
the food sector (Cheetham 1997), the economic be-
nefits of manufacturing natural flavour compounds
will surely remain limited to premium food products
as long as they are significantly higher priced (of-
ten up to two orders of magnitude) than those of
their chemically synthesized analogs. Whereas ini-
tially the tag ‘natural’ was one of the main reasons
for seeking biochemical routes to value-added natural
flavour compounds, now, after decades of research, it
is the accumulated knowledge of the inherent advant-
ages of biocatalysis that is increasingly becoming the
driving force for its application and for the ongoing
research activities in this field. Operation under mild
conditions to protect unstable products, i.e. ‘green’
processes, and functionalization of even complex pre-
cursor molecules in a highly specific way (chemo-,
regio-, stereo-) are two important factors allowing
464
Fig. 1. Synthesis of vanillin from eugenol by genetically engineered Pseudomonas sp. HR 199. Relevant enzymes are indicated; the names of
the corresponding genes are given within the ovals.
biocatalysis to outplay chemical approaches in certain
cases. Research exploiting these advantages of flavour
biocatalysis will certainly be intensified in the future.
Vanillin
Vanillin is by far the most important flavour chemical
in terms of amount and value. It is the main flavour
compound of the cured pods of Vanilla sp. (Ranadive
1994). Annually more than 10 000 tons are produced,
mainly by chemical synthesis. Whereas chemically
produced vanillin is a cheap flavour compound (US$
11 kg−1 ), natural vanillin derived from microbial pro-
cesses currently costs up to US$ 1000 kg−1 and yields
a market volume of several tons annually. Differ-
ent biotechnological production processes are based
on bioconversion of ferulic acid, phenolic stilbenes,
isoeugenol, or eugenol and on de novo biosynthesis,
applying bacteria, fungi, plant cells or genetically en-
gineered microorganisms (Priefert et al. 2001). The
only commercial biocatalytic route is based on the
bioconversion of ferulic acid, frequently occurring in
plants and mainly isolated from rice bran. The vanil-
lin producing strain is Amycolatopsis sp. HR 167.
With this fed-batch process about 12 g product l−1
can be obtained (Rabenhorst 2000). The genes and
enzymes of the producing strain have been identified,
sequenced and cloned (Achterholt et al. 2000). The
reaction is the same as that described for Pseudomonas
sp. HR 199, where the whole pathway from eugenol
over ferulic acid to vanillin has been sequenced (Over-
hage et al. 1999). By disrupting the vdh gene (vanillin
dehydrogenase) this organism can be used for the pro-
duction of vanillin from eugenol, a much cheaper
natural substrate (Figure 1). Since this process in-
volves a genetically modified organism it is currently
not feasible for commercial production in Europe.

Fig. 2. Bioconversion of ricinoleic acid to γ -decalactone by Yarrowia lipolytica.
γ -Decalactone
The most important lactone for flavour application
is γ -decalactone with a market volume of sev-
eral hundred tons per year. It has an oily-peachy
aroma, extraordinarily tenacious odour and a very
powerful, creamy-fruity, peach-like taste in con-
centrations below 5 mg l−1 . In the early eighties
natural γ -decalactone was an extremely expensive,
465
rare natural flavour (>US$ 10 000 kg−1 ). The sub-
sequent introduction and optimization of microbial
processes has resulted in the price dropping to approx.
US$ 300 kg−1 and a market volume of several tons per
year.
Most of the commercial processes use ricinoleic
acid, the main fatty acid of castor oil, or esters
thereof, for the formation of γ -decalactone. The form-
ation of γ -decalactone in the catabolism of ricinoleic
466

Carboxylic acids

Fig. 3. Oxidation of alcohols to carboxylic acids by acetic acid Acetic acid bacteria possess strong oxidative capabil-
bacteria. ities and are used for the oxidation of alcohols to the
corresponding carboxylic acids. With market prices
of < ¤ 100 kg−1 , the carboxylic acids are relatively
acid by yeasts of the genus Candida was first ob-
inexpensive natural flavours but of great importance
served by Okui et al. (1963). Ricinoleic acid is de-
to the flavour industry either because of their intense
graded by four successive cycles of β-oxidation into
smell and sour taste or as substrates for the enzymatic
4-hydroxydecanoic acid which lactonizes at lower pH
production of flavour esters.
to γ -decalactone (Gatfield et al. 1993) (Figure 2). It is
Table 1 summarizes the organoleptic properties of
worth mentioning that the microbial lactone formation
the most important short-chain flavour acids (except
results in the same enantiomeric configuration of the
acetic acid) and data of selected high-yielding biopro-
lactone as is found in peaches and other fruits.
cesses. In the industrial process established by Haar-
The processes with the highest product concentra-
mann & Reimer GmbH (Rabenhorst et al. 2001) the
tions use Yarrowia lipolytica strains. Using a genet-
product yields were increased by optimizing the fer-
ically engineered multiple auxotrophic mutant, desig-
mentation parameters. For all oxidations in the H&R
nated PO1D, Nicaud et al. (1996) obtained high yields
process the molar yield was about 90%.
of γ -decalactone from ricinoleic acid methyl ester.
At the end of the growth phase they transferred the
concentrated biomass into uracil-limited medium for
Esters
biotransformation. After 75 h the culture broth yielded
9.5 g γ -decalactone l−1 (Pagot et al. 1997).
Esters can be found in almost every food and espe-
In a production process established by Haarmann
cially those with a relatively low molecular weight are
& Reimer GmbH (H&R), Germany, up to 11 g γ -
usually fruity in character. The word ‘estery’ is an
decalactone l−1 was obtained in 55 h without a genet-
established term to describe this sensory impression.
ically modified production strain and with raw castor
In plants, esters are formed by lipases or esterases
oil as substrate (Rabenhorst & Gatfield 2000).
starting from amino acids or fatty acids respectively
Other organisms which perform the same reaction
(Belitz & Grosch 1999). These enzymes also play an
with much lower product yields were Monilia fructi-
important role in industrial enzymatic ester formation.
cola (Wink et al. 1988), Sporobolomyces odorus and
Recently, an interesting overview of hydrolase reac-
Rhodotorula glutinis (Cheetham et al. 1988).
tions has been published (Bornscheuer & Kazlauskas
Spinnler’s group studied γ -decalactone produc-
1999).
tion with a number of different Sporidiobolus
Ethyl esters belong to the most important esters
spp. such as Sporidiobolus salmonicolor, Sporidio-
that are relevant for flavours since ethanol is widely
bolus ruinenii, Sporidiobolus johnsonii and Spori-
present in plant metabolism. Methyl, propyl, butyl,
diobolus pararoseus. These strains were very sens-
isobutyl, amyl and isoamyl esters are also of specific
itive to γ -decalactone, while the open form, i.e.
interest for flavours in the food industry. Such natural
4-hydroxydecanoic acid, was less toxic (Dufossé
alcohol residues are derived, for example, from amino
et al. 1998). The yields of γ -decalactone in fer-
acid degradation in nature.
mentations with these yeasts were always very low
Ethyl esters can easily be formed by lipase
(<1 g l−1 ). Cardillo et al. (1989) described a pro-
catalyzed esterification or interesterification reactions
cess using either Aspergillus niger, Pichia etchellsii or
(Figure 4). One great advantage of reactions involving
Cladosporium suaveolens to produce γ -decalactone
lipases or esterases is that they can be carried out in
from castor oil. In all these processes the yields were
organic solvents without the need for any cofactor.
very low (<1 g l−1 ). Kümin & Münch (1997) at
This is of special interest for ester formation since the
Givaudan, Switzerland, used Mucor circillenoides on
water formed can be removed simply by distillation or
ethyl decanoate as substrate for the production of γ -
adsorption.
decalactone. They obtained 10.5 g γ -decalactone l−1
Enzymatic ester reactions can be performed in
after 60 h.
batch or continuous processes. Usually, the enzymes
are used in an immobilized form enabling their applic-
 467
Table 1. Production of carboxylic acids by acetic acid bacteria.

Carboxylic acid Odour/taste; flavouring types Biocatalyst Yield, time Referencea

Propionic acid Pungent, sour, reminiscent of sour milk, Gluconobacter oxydans 94 g l−1 , 92 h 1
cheese, or butter; raspberry, strawberry, Gluconobacter oxydans 44 g l−1 , 70 h 2
cognac, butter Acetobacter pasteurianus 60 g l−1 , 90 h 3
Propionibacterium 20–30 g l−1 , 7–14 d 4
Butyric acid Powerful, penetrating, diffusive sour, remin- Gluconobacter oxydans 95 g l−1 , 72 h 1
iscent of rancid butter; butter, cheese, nut, Acetobacter pasteurianus 60 g l−1 , 90 h 3
fruit, others
Isobutyric acid Powerful, diffusive sour, in dilution almost Gluconobacter oxydans 92 g l−1 , 74 h 1
pleasant and fruity;
cheese, fruit

2-Methylbutyric acid Pungent, acrid, reminiscent of Roquefort Gluconobacter oxydans 80 g l−1 , 72 h 1

cheese, in dilutions pleasant and fruity-sour; Acetobacter pasteurianus 44 g l−1 , 90 h 3
cheese, butter, cream, chocolate

Isovaleric acid Diffusive, acid-acrid, in dilution cheesy, un- Gluconobacter oxydans 82 g l−1 , 72 h 1
(3-Methylbutyric acid) pleasant, in extreme dilution (<20 mg l−1 ) Acetobacter pasteurianus 45 g l−1 , 90 h 3
herbaceous, dry; nut, coffee

The carboxylic acids are produced by oxidation of the corresponding alcohols (except Propionibacterium, which synthesizes the acid de novo).
a [1] Rabenhorst et al. (2001), [2] Svitel & Sturdik (1995), [3] Molinari et al. (1997), [4] Jin & Yang (1998).

Fig. 4. Lipase-catalyzed ester synthesis by esterification or interesterification.

ation in a continuous mode or at least over several pro-

duction batches. Conversion of alcohol and carboxylic
acid or educt ester to the product ester is usually high,
up to 100%.
An example of a more sophisticated ethyl ester
production system is the interesterification of decadi-
enoic acid, derived in situ from stillingia oil, with eth- Fig. 5. Character-impact compounds of pear, ethyl 2(E),4(Z)-
anol to give a character-impact compound for pears, decadienoate, and peppermint, (−)-menthol, biocatalytically
accessible by lipase-catalyzed ester synthesis and hydrolysis, re-
ethyl 2(E),4(Z)-decadienoate (Gatfield et al. 2001)
spectively.
(Figure 5). Since the free acid is not very stable, en-
zymatic in situ interesterification is an elegant way of
circumventing this problem. an important factor since, in nature, many flavour
Esters can be formed by lipases in a racemic or en- compounds do not occur as a racemate but as one
antioselective way. This depends on multiple factors, preferred enantiomer. Moreover, flavour components
such as lipase, enzyme purity, substrate, organic often have a very different sensory profile depending
solvent, and reaction conditions. Enantioselectivity is
468

on their enantioforms (Brenna et al. 2003), which is of

key interest for producing specific products.
One example is enantioselective ester hydrolysis
which can be used to produce enantiopure alcohols.
During the last few years, several groups have de-
veloped enzymatic routes for the production of (−)-
menthol which is by far the most important enantiop-
ure alcohol used in the flavour industry (Figure 5).
One of the latest developments is the enantioselective
cleavage of a racemic menthyl ester [(±)-menthyl ben-
zoate] to yield (−)-menthol with >99% ee and E >
100 (Gatfield et al. 2002). Although produced by an
enzymatic reaction, the flavour compound cannot be
labeled ‘natural’ as the substrate stems from chemical
esterification.

Fig. 6. Ehrlich pathway of yeasts converting amino acids into cor-

2-Phenylethanol responding higher alcohols, here L-phenylalanine into 2-phenyleth-
anol, a rose-like flavour and fragrance compound.

The aromatic alcohol 2-phenylethanol is a flavour and

fragrance compound with a rose-like odour. The over-
all market for 2-phenylethanol was estimated at about
7000 t a−1 in 1990 (Clark 1990) and is dominated
by the chemically synthesized compound. In 2002 an
estimated 0.5 t natural 2-phenylethanol was marketed
(K.-D. Protzen, personal communication) at around
US$ 1000 kg−1 .
Although essential rose oil can – depending on
the variety – contain up to 60% (v/v) of this alco-
hol, it is too valuable to be used as a raw material
for natural 2-phenylethanol which is required for food
flavourings. Industrial processes use recovery from
distillation residues (Savina et al. 1999) or microbial
fermentation (www.safisis.com). Research is continu-
ing to improve the economics of bioprocesses which
may result in an expansion of the market share of
natural 2-phenylethanol.
Although certain bacteria produce 2-phenylethanol
(Jollivet et al. 1992, Spinnler et al. 1991), research
on microbial production favours yeasts as biocata-
lysts. Pathways leading to 2-phenylethanol are sum-
marized elsewhere (Etschmann et al. 2002), the most
important being the Ehrlich pathway (Figure 6): L-
phenylalanine is transaminated to phenylpyruvate, de-
carboxylated to phenylacetaldehyde and finally re-
duced to 2-phenylethanol.
By supplementing the medium with L-phenyl-
alanine, the Ehrlich pathway is accelerated and high
product concentrations can be achieved (Etschmann
et al. 2003a). These cultures, however, are product
inhibited by 2-phenylethanol, thus in-situ product re-
moval is essential. Of the different options tested
(Fabre et al. 1996), mainly liquid-liquid extraction
has found further interest in research. Two-phase
fermentation of Kluyveromyces marxianus CBS 600
with oleyl alcohol in shake flasks yielded 3 g 2-
phenylethanol l−1 (Etschmann et al. 2003a); this
has been increased to 5.6 g l−1 by medium optim-
ization using a genetic algorithm (Etschmann et al.
2004). In an optimized fed-batch procedure using Sac-
charomyces cerevisiae with oleic acid as extractant,
12.6 g 2-phenylethanol l−1 was obtained although syn-
ergistic inhibitory effects of extractant and product
were still observed (Stark et al. 2002). Inhibitory ef-
fects of dispersed organic phases can be circumvented
by in-situ product removal techniques based on mem-
branes, e.g. microbial production of 2-phenylethanol
in a bioreactor coupled with an organophilic perva-
poration module has already been demonstrated to
be an effective technique (Etschmann et al. 2003b)
and is currently being investigated on pilot scale with
promising results.
C6 aldehydes and alcohols (‘green notes’)
The industrially most relevant compounds to obtain
the ‘green’ organoleptic characteristic are C6 alde-
hydes and their corresponding alcohols (Figure 7).
For example, 3(Z)-hexen-1-ol (‘leaf alcohol’) has a
powerful odour of freshly cut grass and is an important

Fig. 7. Biocatalytic routes to ‘green notes’ from linolenic and linoleic acid.
flavour and fragrance material used for natural green
top notes (Bauer et al. 2001).
The natural green notes market is estimated at
5–10 t a−1 and US$ 3000 kg−1 (Muller et al.
1995a). Since the traditional preparation of nat-
ural green notes by distilling plant oils, e.g. mint
terpene fractions, is costly and cannot meet the in-
creasing market demand for natural flavours, dif-
ferent biocatalytic syntheses have been developed,
all containing at least one enzymatic reaction cata-
lyzed by crude plant material (Goers et al. 1989,
Brunerie & Koziet 1997, Belin et al. 1998, Holtz
et al. 2001). In comparison with these patents, one
filed by Firmenich (Muller et al. 1995b) reported
the highest yields, e.g. 4.2 g 3(Z)-hexen-1-ol kg−1
469
and 1.5 g 2(E)-hexenal kg−1 were produced. The
biocatalysis starts from linoleic or linolenic acid
and comprises a sequence of three enzymatic steps:
the lipoxygenase-catalyzed functionalization of li-
noleic acid and linolenic acid to 13(S)-hydroperoxy-
octadeca-9(Z),11(E)-dienoic acid (‘C13-HPOD’)
and 13(S)-hydroperoxy-octadeca-9(Z),11(E),15(Z)-
trienoic acid (‘C13-HPOT’), respectively; the cleavage
of the C13-hydroperoxide acids to release the desired
C6 aldehydes by the action of hydroperoxide lyase (a
cytochrome P450 enzyme); the reduction of the C6
aldehydes to their corresponding alcohols by alcohol
dehydrogenase.
As the use of isolated enzymes would render the
biocatalytic approaches uneconomic, crude biological
470
materials and baker’s yeast have been combined to
form a modular synthetic system. Soya flour, by the
action of lipoxygenase, catalyzes the oxidation of un-
saturated fatty acids, which may be used as hydrolyzed
oils of sunflower or linseed dissolved in water. The
identification of a catalytically active plant material
for the hydroperoxide lyase step was crucial to the
overall economics of the process, as no commercially
available hydroperoxide lyase has been developed so
far and hydroperoxide lyase activities in plant tissue
are usually low. Guava fruit homogenate best fulfilled
the technical and economic requirements. In the case
of C6 alcohols as target green notes, a third catalytic
step is performed by dosing active baker’s yeast as
the reductive reagent. During the production of unsat-
urated green notes, catalysis by hydroperoxide lyase
and reductase at the same time primarily causes an
accumulation of 3(Z)-hexenol; this is because its pre-
cursor 3(Z)-hexenal is an unstable intermediate which
quickly undergoes non-enzymatic conversion to 2(E)-
hexenal. Thus, a lagged dosage of the final reductase
entails 2(E)-hexenol as the predominant green alco-
hol, whereas ending the process by immediate solvent
extraction after the lyase step leads to 3(Z)-hexenal.
In contrast, performing the isomerization but not the
reduction leads to 2(E)-hexenal as the main product.
Therefore, by varying the catalytic modules and start-
ing materials, all members of the C6 green note family
can be produced.
Since the lyase reaction is rate-limiting, a novel
production strategy based on genetically modified
Saccharomyces cerevisiae coexpressing the hydroper-
oxide lyase gene from banana and the lipoxygenase
gene, thus unifying all enzyme activities needed in one
host, has been reported (Muheim et al. 1997, Häusler
et al. 2001).
Future prospects
Innovative strategies for biocatalytic flavour gener-
ation will certainly take advantage of the immense
progress currently being made in the emerging fields
of functional genomics, proteomics, protein and meta-
bolic engineering. As an increasing number of genome
sequence data becomes available, the improved ge-
nomic and proteomic methodologies provide access
to new enzymes including those playing key roles
in plant flavour biosynthesis. Modern microbial host
vector systems simplify the functional expression and
molecular and biochemical characterization of hetero-
logous enzymes. An increasing amount of research
work is being published which gives insight into plant
biosynthesis by ‘isolation, cloning and expression’
approaches.
Croteau and co-workers elucidated the enzymatic
catalysis and stereochemistry of (−)-limonene hy-
droxylations in different mint species by heterologous
expression in E. coli and S. cerevisiae (Haudenschild
et al. 2000, Wüst et al. 2001). In peppermint (Mentha
x piperita) (−)-limonene is converted into (−)-trans-
isopiperitenol by a P450 monooxygenase regiospe-
cifically oxyfunctionalizing the C3 position, the first
reaction of a subsequent cascade of five enzymes lead-
ing to the important flavour molecule (−)-menthol.
By functional expression in E. coli, a multifunc-
tional O-methyltransferase, catalyzing the formation
of 2,5-dimethyl-4-methoxy-3(2H)-furanone (DMMF),
an impact compound in strawberry flavour, has suc-
cessfully been characterized (Wein et al. 2002). These
are only a few examples among many others to illus-
trate the progress currently being made by genetic en-
gineering in flavour biosynthesis research. Functional
expression of newly discovered, flavour-generating
plant enzymes in bacteria or yeasts constitutes a po-
tential starting point for a bioprocess-oriented optim-
ization, but only the establishment of high-level ex-
pression systems can transfer them to real whole-cell
biocatalysts.
Additionally, molecular biological methodologies
are necessary to optimize biocatalyst properties, e.g.
by conferring higher stereoselectivity or thermostabil-
ity or even a different substrate spectrum. The active
site of Pseudomonas putida heme monooxygenase
P450cam , which naturally converts (+)-camphor to 5-
exo-hydroxycamphor, was remodeled by site-directed
mutagenesis and a double mutant showed completely
different substrate and product spectra (Bell et al.
2001): (−)-limonene and α-pinene were hydroxylated
with high regio- and stereoselectivities to (−)-trans-
isopiperitenol and (+)-cis-verbenol, respectively. A
strategy of potential future impact on natural flavour
production addresses the total synthesis of terpenoids
by engineering the mevalonate-dependent isoprenoid
pathway comprising eight genes from S. cerevisiae
into E. coli (Martin et al. 2003). The goal is a bac-
terial cell factory providing the universal precursors of
terpenoid flavour compounds, isopentenyl pyrophos-
phate and dimethylallyl pyrophosphate, by de novo
synthesis. This might grant access to the variety of
terpenoid flavours once the appropriate genes coding

terpene synthases and functionalizing enzymes have
been heterologously expressed in the same host strain.
The feasibility of novel biocatalytic flavour syn-
theses will not only depend on genetically improved
biocatalysts but also on process engineering features.
This is especially evident in the case of terpenoid
flavour compounds, an area of high commercial in-
terest to the industry due to the large number of key
aroma compounds which are in principle accessible by
biotransformation of abundant natural terpene hydro-
carbons (Schrader & Berger 2001). Biotransformation
attempts have to overcome several drawbacks, such
as the low water-solubility of the precursors, toxicity
of precursors and products, and metabolic diversity
which leads to unwanted by-products or further de-
gradation of the target molecules. One example which
shows that extremely high yields can nevertheless be
obtained was given by Fontanille & Larroche (2003).
Up to 400 g 2(Z)-methyl-5-isopropyl-2,5-hexadienal
(isonovalal) l−1 , an artificial fragrance compound for
potential use in perfume formulations, was produced
from α-pinene oxide within 2.5 h using 25 g pre-
cultivated biomass l−1 (Pseudomonas rhodesiae CIP
107491); the cells had been permeabilized by freeze-
thawing and organic solvent treatment prior to use.
The bioprocess was performed with in situ product
recovery using hexadecane in a biphasic medium and
by sequential feeding of biomass and precursor to
compensate the irreversible biocatalyst inactivation by
the product. Recently, enzyme stabilization, a very
important issue for in vitro biocatalysis, has success-
fully been addressed using sol-gel encapsulation of
both a P450 monooxygenase and a NADP+ -dependent
formate dehydrogenase for cofactor-recycling (Maurer
et al. 2003). By this means, β-ionone was regiose-
lectively hydroxylated at the C4 position, a reaction of
importance for the generation of tobacco flavourings.
Nevertheless, how long it will take before novel
biocatalysis strategies lead to industrially applied pro-
cesses in Europe will not only depend on their sci-
entific and technological feasibility but also, if not
exclusively, on a less prejudiced and more balanced
public perception of the use of genetic engineering
for improved food quality and more environmentally
friendly production processes.
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