Scope and limitations of biocatalytic carbonyl reduction with white-rot fungi

Article abstract of DOI:10.1016/j.bioorg.2021.104651

The reductive activity of various basidiomycetous fungi towards carbonyl compounds was screened on an analytical level. Some strains displayed high reductive activities toward aromatic carbonyls and aliphatic ketones. Utilizing growing whole-cell cultures of Dichomitus albidofuscus, the reactions were up-scaled to a preparative level in an aqueous system. The reactions showed excellent selectivities and gave the respective alcohols in high yields. Carboxylic acids were also reduced to aldehydes and alcohols under the same conditions. In particular, benzoic, vanillic, ferulic, and p-coumaric acid were reduced to benzyl alcohol, vanillin, dihydroconiferyl alcohol and 1-hydroxy-3-(4-hydroxyphenyl)propan, respectively.

Full text of DOI:10.1016/j.bioorg.2021.104651

Bioorganic Chemistry 108 (2021) 104651
Contents lists available at ScienceDirect
Bioorganic Chemistry
journal homepage: www.elsevier.com/locate/bioorg
Scope and limitations of biocatalytic carbonyl reduction with
white-rot fungi
,
b
,
,
,c
Tatyana S. Zhuka *, Oleksandra S. Skorobohatkoa b, Wendell Albuquerquea, Holger Zorna
a Institute of Food Chemistry and Food Biotechnology, Justus Liebig University Giessen, Heinrich-Buff-Ring, 17, 35392 Giessen, Germany
b Department of Organic Chemistry, Igor Sikorsky Kyiv Polytechnic Institute, Peremogy Ave., 37, 03056 Kyiv, Ukraine
c Fraunhofer Institute for Molecular Biology and Applied Ecology, Ohlebergsweg 12, 35392 Giessen, Germany
A R T I C L E I N F O
A B S T R A C T
Keywords:
The reductive activity of various basidiomycetous fungi towards carbonyl compounds was screened on an
analytical level. Some strains displayed high reductive activities toward aromatic carbonyls and aliphatic ke-
tones. Utilizing growing whole-cell cultures of Dichomitus albidofuscus, the reactions were up-scaled to a pre-
parative level in an aqueous system. The reactions showed excellent selectivities and gave the respective alcohols
in high yields. Carboxylic acids were also reduced to aldehydes and alcohols under the same conditions. In
particular, benzoic, vanillic, ferulic, and p-coumaric acid were reduced to benzyl alcohol, vanillin, dihy-
droconiferyl alcohol and 1-hydroxy-3-(4-hydroxyphenyl)propan, respectively.
Whole-cell cultures
Bio-reduction
White-rot fungi
Carbonyls
D. albidofuscus
1. Introduction
peroxygenases such as chloroperoxygenases (CPO) [13] and unspecific
peroxygenases (UPO) [14], class II heme-containing peroxidases, and
Hydride-containing agents traditionally employed for reducing
carbonyl compounds currently raise increasing environmental concerns
[1]. Alternative catalytic hydrogenations require heavy metal-based
catalysts and often harsh reaction conditions [2]. High selectivities,
mild reaction conditions and environmental safety attract the attention
to biocatalytic reductions as alluring alternative to traditional organic
synthesis, especially in the food, pharmaceutical and fine chemical in-
dustries [3–6]. Nicotinamide adenine dinucleotide (NAD(P)H) depen-
dent dehydrogenases are the catalysts of choice for the reduction of
carbonyls [7]. The best-investigated sources of reductive enzymes are
yeasts and bacteria, while the reductive potential of basidiomycetous
fungi was disclosed only on an analytical level and for a very limited
number of cultures so far [8].
various copper-containing oxidoreductases including laccases [15,16]
are the most active components of the extracellular fungal secretome
and have been studied extensively [9,17–21]. Lignolytic enzymes
responsible for intracellular degradation of low-molecular weight frag-
ments of lignin have been the subject of intense recent studies [12].
Numerous attempts to utilize basidiomycetous enzymes both in isolated
form and as whole-cell biocatalysts for various types of oxidations have
been thoroughly reviewed [14,15,22]. The first reduction catalysed by a
basidiomycetous fungus, Trametes versicolor, has been described as early
as in 1959 for the transformation of aromatic carboxylic acids to the
corresponding aldehydes and alcohols [23,24]. Various fungi were
screened for their ability to reduce carboxylic acids of different nature,
namely fluorinated benzoic, isomeric anisic, and amine containing
benzoic acid derivatives on an analytical level [8,25,26]. The use of
isolated enzymes currently dominates in preparative enzymatic catalysis
[27–30] due to the high selectivities and yields. However, costly puri-
fication processes, cofactor regeneration steps, presence of auxiliary
enzymes as well as the low stability of many isolated enzymes represent
significant drawbacks [30].
In the current study, we focused on enzymes produced by white-rot
fungi which represent versatile natural catalysts due to their unique
ability to degrade lignin, the most abundant aromatic biopolymer on
earth [9–11]. Lignin is characterized by a highly complex three-
dimensional structure based on p-coumaryl, coniferyl, and sinapyl
moieties [11]. Its degradation into low-molecular weight aromatic
compounds is a complex process catalysed by extracellular enzymes and
followed by intracellular metabolic bioconversions [12]. Heme-thiolate
All these disadvantages may be overcome when whole-cell systems
are utilized. They are also simpler in application and can serve as
* Corresponding author at: Institute of Food Chemistry and Food Biotechnology, Justus Liebig University Giessen, Heinrich-Buff-Ring, 17, 35392 Giessen,
Germany.
E-mail addresses: t.zhuk@xtf.kpi.ua, tetiana.zhuk@lcb.chemie.uni-giessen.de (T.S. Zhuk).
https://doi.org/10.1016/j.bioorg.2021.104651
Received 3 November 2020; Accepted 7 January 2021
Available online 12 January 2021
0045-2068/© 2021 Elsevier Inc. All rights reserved.

T.S. Zhuk et al.
Bioorganic Chemistry 108 (2021) 104651
Fig. 1. Model compounds benzaldehyde (1), p-nitrobenzaldehyde (2), cyclohexanone (3), adamantanone (4) utilized for screening procedure (A) and the workflow
of the screening (B, MC = main culture).
efficient “green” catalysts [31]. On the other hand, fungal secretomes
consist of a complex mixture of enzymes of different nature that are able
to catalyse a broad range of chemical transformations, and are thus not
expected to be selective if whole-cell cultures are utilized. Nevertheless,
examination of whole-cell transformations is typically the first step in
revealing the catalytic properties of enzymes.
purchased from Merck KGaA (Darmstadt, Germany).
2.2. Organisms
The stock culture of the fungi Dichomitus albidofuscus (CBS, 321.75),
Marasmius cohortalis (DSMZ, 8257), Pleurotus sapidus (DSMZ, 8266),
Flammulina velutipes (DSMZ, 1658), Daedalea quercina (DSMZ, 4953),
Ischnoderma benzoinum (CBS, 231.97), Pleurotus cornucopiae (DSMZ,
5342), Pleurotus ostreatus (DSMZ, 1833), and Dichomitus squalens (DSMZ,
9615) were maintained on a solid medium containing 15 g⋅Lꢀ 1 malt
extract (Fluka, Neu-Ulm, Germany) and 15 g⋅Lꢀ 1Agar-Agar (Roth,
Karlsruhe, Germany).
In this study, we reveal the reductive potential of whole-cell cultures
of several white-rot fungi. In particular, Dichomitus squalens was chosen
as this fungus has been shown to degrade chlorophenoxyacetic acids
[32], and its carboxylate reductase was recently characterized [33].
Additionally, Dichomitus albidofuscus, which is able to oxidase aliphatic
compounds [34], Ischnoderma benzoinum that transforms L-phenylala-
nine to benzaldehyde [35], and Daedalea quercina whose laccases are
able to decolorize synthetic dyes efficiently [36] were selected for the
present study. Whereas the chemistry of Daedalea quercina and Mar-
asmius cohortalis remains largely undisclosed, fungi of the Pleurotus
family, namely P. sapidus, P. ostreatus, and P. cornucopiae, display an
enormously rich chemistry and various applications in biotechnology
[20,37–39] and are also subject of our scrutiny.
2.3. Submerged cultures
The culture medium was prepared by dissolving malt extract (30 g)
in 1 L of deionised water. For preparation of the precultures, a 1 cm2
agar plug from the leading mycelial edge was transferred into 100 mL
medium (250 mL Erlenmeyer flask) and then homogenized with a T 25
digital Ultra-Turrax homogenizer (IKA, Staufen, Germany; 30 s, 10.000
r⋅minꢀ 1). The precultures were grown on an incubation shaker (Orbi-
tron, Infors HAT, Bottmingen, Switzerland; 150 r⋅minꢀ 1, deflection 25
mm) under exclusion of light at 24 ◦C for 7 days. Afterwards, the pre-
cultures were homogenized, and 10% (v/v) of the homogenate was
inoculated for submerged cultivation into 400 mL (1000 mL Erlenmeyer
flask), 200 mL (500 mL Erlenmeyer flask) or 40 mL (100 mL Erlenmeyer
flask) medium.
2. Materials and methods
2.1. Chemicals
Benzaldehyde, benzoic acid, cyclohexanone, adamantanone, 3-
chloro-4-methoxybenzaldehyde,
m-methylbenzaldehyde,
m-nitrobenzaldehyde,
vera-
traldehyde,
m-anisaldehyde,
p-nitro-
benzaldehyde,
p-bromobenzaldehyde,
2-heptenal,
dodecanal,
undecanal, m-nitrobenzoic acid, p-nitrobenzoic acid, vanillic acid, cin-
namonic acid, ferulic acid, p-coumaric acid (for synthesis), benzyl
alcohol, cyclohexanol, 2-adamantanol, 3-chloro-4-methoxybenzyl
alcohol, m-methylbenzyl alcohol, veratryl alcohol, m-anisyl alcohol, m-
nitrobenzyl alcohol, p-nitrobenzyl alcohol, p-bromobenzyl alcohol, 1-
heptanol, 1-dodecanol, 1-undecanol, vanillin, dihydroconiferyl
alcohol, 1-hydroxy-3-phenylpropanol-1-ol, 1-hydroxy-3-(4-hydrox-
yphenyl)propan (as standards for compound identification) were
2.4. Screening procedure
The substrate (0.5 mmol) was added to submerged cultures of the
respective fungus (40 mL) on the 3rd culture day. The reaction mixture
was incubated on an incubation shaker at 150 r⋅minꢀ 1 (deflection 25
mm) under exclusion of light at 24 ◦C for 4 days. Afterwards, 5 g of NaCl
was added to the medium, and the mixture was stirred for 10 min at 800
rpm (magnetic stirrer). For extraction 25 mL of Et2O was added, and the
2

T.S. Zhuk et al.
Bioorganic Chemistry 108 (2021) 104651
Fig. 2. Screening results for biotransformation of benzaldehyde (1, A), p-nitrobenzaldehyde (2, B), cyclohexanone (3, C), adamantanone (4, D) in the presence of
various fungi.
resulting mixture was stirred for 20 min at 800 rpm (magnetic stirrer),
and centrifuged for 5 min at 3000 × g to separate the organic layer. The
extraction was repeated three times. The combined organic layers were
washed with brine (1 × 30 mL) and water (1 × 30 mL), dried over
Na2SO4 and evaporated to dryness. The resulting reaction mixtures were
analysed by GC–MS (Agilent Technologies 7890A GC, column Agilent
VF-WAXms (30 m × 0.25 mm, 0.25 µm) and Agilent 5975C MSD Triple-
Axis mass spectrometer) and GC-FID (Agilent Technologies 7890A GC,
7683B Series Injector, column Agilent HP5 30 m × 0.2 mm, 0.25 µm).
Every biotransformation was repeated three times to verify the repro-
ducibility of the experiments. The detailed information about experi-
mental data and yields is presented in the Electronic Supplementary
Information.
experiments. The detailed information about experimental data and
yields is provided in the Electronic Supplementary Information.
3. Results and discussion
3.1. Screening of reductive activity of selected fungi
Several basidiomycetous fungi were screened for their potential to
reduce benzaldehyde (1) and p-nitrobenzaldehyde (2) (Fig. 1, A) as
substituted aromatics to a certain extent mimic the lignin fragments
[40]. Compound 2 was chosen because of the electron-withdrawing
nature of the nitro-group. As mentioned above, whole-cell cultures
usually contain a complex mixture of enzymes including highly active
oxidative species, especially laccases and peroxidases that are even able
to oxidize non-functionalized aliphatic hydrocarbons [34]. Therefore,
the aliphatic ketones cyclohexanone (3) and adamantanone (4) that may
potentially be oxidized or reduced by the fungal enzymes were included
in the screening. For the screening, the fungi were grown in submerged
cultures for three days prior to the addition of the model compounds 1–4
(Fig. 1, B).
2.5. General procedure for preparative biotransformation
The substrate (1 mmol) was added to submerged cultures of DAL on
the 3rd culture day. The reaction mixture was incubated on an incuba-
tion shaker at 150 r⋅minꢀ 1 (deflection 25 mm) under exclusion of light at
24 ◦C for 4 days. 10 g of NaCl was added to media, and the mixture was
stirred for 10 min at 800 rpm (magnetic stirrer). For extraction, 50 mL of
Et2O was added, and the resulting mixture was stirred for 20 min at 800
rpm (magnetic stirrer), and centrifuged for 5 min at 3000 × g to separate
the organic layer. The extraction was repeated three times. The com-
bined organic layers were washed with brine (1 × 30 mL) and water (1
× 30 mL), dried over Na2SO4 and evaporated to dryness. The resulting
reaction mixtures were analysed by GC/MS and NMR spectroscopy.
Products were purified by column chromatography on silica gel (eluent
(pentane/ether) changed gradually: 10/1, 7/3, 1/1) to isolate the major
components. The respective fractions were combined, concentrated in
vacuum, and the 1H and 13C NMR spectra of the residuals were
compared with those of reference compounds. Every biotransformation
was repeated three times to verify the reproducibility of the
The results of the biotransformation experiments differed for each
compound. (Fig. 2) For substrate 1, benzylic alcohol (5) and benzoic
acid (6) were found as products, while for substrate 2 only the reduced
product 7 was detected. The aliphatic ketone 3 was solely reduced to the
corresponding alcohol, while for compound 4 a complex mixture of CH-
oxidation products 10 was detected apart from the reduction product 9,
especially with M. cohortalis.
The only fungus that did not show any reduction of 1 or 2 was
P. ostreatus. All fungi, except for P. sapidus and I. benzoinum were able to
reduce benzaldehyde (1) to benzyl alcohol (2) (Fig. 2, A). For 1 the
reduction often competes with oxidation to benzoic acid (6), and only
with D. squalens and D. albidofuscus solely the reduction product 5 was
formed in high yields. No traces of p-nitrobenzoic acid as well as
3

T.S. Zhuk et al.
Bioorganic Chemistry 108 (2021) 104651
Table 1
Preparative reduction of aldehydes with growing whole-cell cultures of Dichomitus albidofuscus (DAL).
derivatives with reduced nitro group have been detected in any reaction
mixture. All fungi except P. ostreatus reduced cyclohexanone (3) effi-
ciently to cyclohexanol (8) without any traces of oxidative products.
Biotransformations with adamantanone (4) gave the most complex
product spectrum. Apart from 2-hydroxyadamantane (9), hydrox-
yadamantanone and dihydroxyadamantane (10) were detected with
fungi of the Pleurotus family. The reaction mixtures obtained from cul-
tures of M. cohortalis contained only disubstituted derivatives of ada-
mantane 10.
Compared to other fungi, D. albidofuscus (DAL) demonstrated the
highest reduction efficiency towards all of the tested model substrates
(Fig. 2). Apart from that, only moderate amounts of biomass were
4

T.S. Zhuk et al.
Bioorganic Chemistry 108 (2021) 104651
Fig. 3. 13C NMR spectra of the extracts from the reaction mixtures (RM) of the reduction with the D. albidofuscus for 3-chloro-4-methoxybenaldehyde (A), 3-heptenal
(B), m-nitrobenzoic acid (C) and p-nitrobenzoic acid (D). All resonances correspond to the signals of inset structures.
formed during the culture period which simplified the workup proced-
ure. This fungus was thus selected for preparative experiments on the
reduction of various classes of carbonyl compounds.
18 can serve as a substrates for extracellular aryl-alcohol oxidases that
reduce oxygen to hydrogen peroxide [41]. The carbonyl groups of the
isomeric nitrobenzaldehydes 19 and 2 were reduced to the corre-
sponding alcohols 20 and 7 after 3 days almost quantitatively.
p-Bromobenzaldehyde (21) was transformed to alcohol 22 with
3.2. Reduction of aldehydes with D. albidofuscus
moderate yields. We also tested the reduction of α-β-unsaturated alde-
hyde, namely 2-heptenal (23), that was transformed into the corre-
sponding saturated alcohol 24 with high yields. The enzymatic
reduction of activated double bonds has been described for the old
yellow enzyme family of flavoprotein oxidoreductases [42], where the
For studying the scope and limitations of the reduction with DAL, a
number of substituted aromatic and aliphatic aldehydes (Table 1) were
supplemented to its submerged cultures on the 3rd culture day. All re-
actions were performed with 1 mmol of substrates, and the crude ex-
tracts were analysed by GC–MS in combination with 1H- and 13C NMR
spectroscopy in order to estimate the actual amount of non-volatile
impurities. The products were than isolated from the reaction mix-
tures by column chromatography and compared to authentic reference
compounds. The whole-cell cultures reduced 3-chloro-4-methoxyben-
zaldehyde (11) to the corresponding alcohol 12 with a preparative
yield of about 80% (Table 1). Comparable observations were performed
with m-methylbenzaldehyde (13), and the yield of the corresponding
alcohol 14 was ca. 50%. Veratraldehyde (15) gave the alcohol 16 (31%),
and m-anisaldehyde (17) gave 49% of the corresponding alcohol 18. The
moderate yields here could be attributed to the fact that alcohols 16 and
–
carbonyl group is maintained and only reduction of the C C double
–
bond is observed. The linear saturated aldehydes 25 and 27 dodecanal
and undecanal were readily reduced to the corresponding alcohols 26
and 28 with high yields by DAL.
In many cases, the selectivity of the whole-cell cultures of DAL re-
ductions was outstanding. We conclude this based on the NMR-analyses
of crude extracts from the reaction mixtures where the target products
clearly dominated as deduced from the spectral data of typical crude
reaction mixtures (Fig. 3). The main resonances of the 13C NMR spectra
of the reaction mixtures of reduction of 3-chloro-4-methoxybenzalde-
hyde (11, Fig. 3A) and 2-heptenal (23, Fig. 3B) agreed well with the
Fig. 4. Preparative biotransformation of adamantanone (4) by D. albidofuscus.
5

T.S. Zhuk et al.
Bioorganic Chemistry 108 (2021) 104651
Fig. 5. Reduction of carboxylic acids by growing whole-cell cultures of D. albidofuscus.
spectral data of the corresponding standard samples. Despite the whole-
cell culture obviously produced a large set of enzymes of different na-
ture, the preparative reductions of aldehydes by DAL are highly selective
and only trace amounts of oxidation products, namely the corresponding
carboxylic acids, have been found in the reaction mixtures.
corresponding alcohols 12 and 7 with high yields after addition of the
substrate on the 2nd culture day (Fig. 3, C and D). While the carboxylic
acid and (intermediate) aldehyde functions were reduced, the nitro
group remained unchanged. Vanillic acid (31) was reduced under the
same conditions to vanillin (32) with 35% preparative yield.
DAL cultures were also employed for the reduction of cinnamic acid
(33) and its substituted derivatives containing coniferyl and p-coumaryl
moieties. During three days, the double bonds and carboxyl groups of
acids 35 and 37 were reduced giving dihydroconiferyl alcohol (36) and
1-hydroxy-3-(4-hydroxyphenyl)propan (38), respectively (Fig. 5). Only
traces of alcohol 34 were found in the reaction mixture of trans-
formation of cinnamic acid (33) under the same conditions.
Summarizing the results obtained for acids, we conclude that
growing whole-cell cultures of DAL may be efficiently employed for
reductive processes. Moreover, the fungus is able to reduce aromatic
acid derivatives to aldehydes and alcohols with satisfactory preparative
yields.
3.3. Reduction of adamantanone with D. albidofuscus
The biotransformation of admantanone (4) with DAL gave the
alcohol 9 with an isolated yield of 71%. (Fig. 4) Traces of disubstituted
derivatives of adamanatane which accumulated at prolonged reaction
times were detected in the reaction mixture.
To localize the enzymes responsible for the reductions, biotransfor-
mation experiments were performed separately with supernatants and
mycelia of 4 days old cultures. While no traces of alcohol 9 were found in
the reaction mixtures containing culture supernatants, 22% of ada-
mantanol (9) accumulated after 24 h when the reaction was performed
with mycelium. The responsible enzymes are thus likely located
intracellularly.
4. Conclusions
Basidiomycetous fungi are able to selectively reduce carbonyl groups
not only of aromatic compounds but also of saturated ketones and al-
dehydes. Growing cultures of D. albidofuscus display remarkable selec-
tivities in preparative “on water” reductions of aromatic carbonyl
compounds with high yields at room temperature. The fact that this
fungus is able to reduce benzoic acids to alcohols with high preparative
yields is extraordinary as such transformations usually require either
hazardous organic reagents or harsh reaction conditions in organic
chemistry. Some functional groups resist the reaction conditions which
may be useful for the reduction of polyfunctional aromatics. Detailed
mechanistic studies as well as isolation of responsible intermediates in
whole-cell culture of D. albidofuscus chemistry are currently underway.
3.4. Reduction of carboxylic acids with D. albidofuscus
The reduction of carboxylic acids to the respective alcohols usually
requires either prolonged heating with large amounts of hydride re-
agents [43–45] or involvement of heavy-metal-based catalysts [46].
With DAL, benzoic acid (6) was readily reduced to benzyl alcohol (5)
during 3 days with 82% yield when 6 was added on the 2nd culture day
(Fig. 5). In contrast, when 6 was added on the 6th culture day, only 1
was found in the reaction mixture and isolated in 45% preparative yield.
It should be noted that this approach gives halogen-free benzaldehyde
(1) which is essential for applications in the pharmaceutical and food
industry [47].
m-(29) and p- (30)Nitrobenzoic acids were readily reduced to the
6

T.S. Zhuk et al.
Bioorganic Chemistry 108 (2021) 104651
˜
[18] A.T. Martínez, F.J. Ruiz-Duenas, S. Camarero, A. Serrano, D. Linde, H. Lund,
J. Vind, M. Tovborg, O.M. Herold-Majumdar, M. Hofrichter, C. Liers, R. Ullrich,
K. Scheibner, G. Sannia, A. Piscitelli, C. Pezzella, M.E. Sener, S. Kılıç, W.J.H. van
Declaration of Competing Interest
The authors declare that they have no known competing financial
interest or personal relationships that could have appeared to influence
the work reported in this paper.
´
Berkel, V. Guallar, M.F. Lucas, R. Zuhse, R. Ludwig, F. Hollmann, E. Fernandez-
Fueyo, E. Record, C.B. Faulds, M. Tortajada, I.b. Winckelmann, J.-A. Rasmussen,
´
M. Gelo-Pujic, A. Gutierrez, J.C. del Río, J. Rencoret, M. Alcalde, Oxidoreductases
on their way to industrial biotransformations, Biotechnol. Adv. 35 (6) (2017)
815–831, https://doi.org/10.1016/j.biotechadv.2017.06.003.
´
[19] A. Hernandez-Ortega, P. Ferreira, A.T. Martínez, Fungal aryl-alcohol oxidase: a
Acknowledgements
peroxide-producing flavoenzyme involved in lignin degradation, Appl. Microbiol.
Biotechnol. 93 (4) (2012) 1395–1410, https://doi.org/10.1007/s00253-011-3836-
8.
TSZ is grateful to German Academic Exchange Service (DAAD) for
research fellowship (# 57314018) and to Humboldt Foundation for
Georg Forster Research Fellowship. The work was supported in part by
the State Fund for Fundamental Research (project # F83/73-2018). OSS
is grateful to Giessen University for the Liebig Fellowship. This project
has partially been financed with funds of LOEWE – Landes-Offensive zur
[20] H. Zorn, T. Peters, M. Nimtz, R.G. Berger, The secretome of Pleurotus sapidus,
Proteomics 5 (18) (2005) 4832–4838, https://doi.org/10.1002/(ISSN)1615-
986110.1002/pmic.v5:1810.1002/pmic.200500015.
[21] H. Bouws, A. Wattenberg, H. Zorn, Fungal Secretomes – Nature’s toolbox for white
biotechnology, Appl. Microbiol. Biotechnol. 80 (2008) 381–388, https://doi.org/
10.1007/s00253-008-1572-5.
[22] C. Schmidt-Dannert, Biocatalytic portfolio of Basidiomycota, Curr. Opin. Chem.
Biol. 31 (2016) 40–49, https://doi.org/10.1016/j.cbpa.2016.01.002.
[23] V.C. Farmer, M.E.K. Henderson, J.D. Russell, Reduction of certain aromatic acids to
aldehydes and alcohols by Polystictus versicolor, Biochim. Biophys. Acta, Proteins
Proteomics 35 (1959) 202–211, https://doi.org/10.1016/0006-3002(59)90349-x.
[24] H. Shimazono, F.F. Nord, Transformations of anisic acid and methylanisate by the
mold Polystictus versicolor, Arch. Biochem. Biophys. 87 (1) (1960) 140–143,
https://doi.org/10.1016/0003-9861(60)90136-3.
¨
Entwicklung Wissenschaftlich-okonomischer Exzellenz-AromaPlus
(State Offensive for the Development of Scientific and Economic
Excellence).
Appendix A. Supplementary material
[25] M. Winkler, Carboxylic acid reductase enzymes (CARs), Curr. Opin. Chem. Biol. 43
(2018) 23–29, https://doi.org/10.1016/j.cbpa.2017.10.006.
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.bioorg.2021.104651.
[26] K. Napora-Wijata, G.A. Strohmeier, M. Winkler, Biocatalytic reduction of
carboxylic acids, Biotechnol. J. 9 (6) (2014) 822–843, https://doi.org/10.1002/
biot.201400012.
[27] E.G. Hibbert, P.A. Dalby, Directed evolution strategies for improved enzymatic
performance, Microb. Cell Factories 4 (2005), https://doi.org/10.1186/1475-
2859-4-29.
References
[1] J. Magano, J.R. Dunetz, Large-scale carbonyl reduction in the pharmaceutical
industry, Org. Process Res. Dev. 16 (2012) 1156–1184, https://doi.org/10.1021/
op2003826.
[28] S.E. Milner, A.R. Maguire, Recent trends in whole cell and isolated enzymes in
enantioselective synthesis, Arkivoc 2012 (2012) 321–382, https://doi.org/
10.3998/ark.5550190.0013.109.
[2] F.L. Bernardis, R.A. Grant, D.C. Sherrington, A review of methods of separation of
the platinum-group metals through their chloro-complexes, React. Funct. Polym.
65 (3) (2005) 205–217, https://doi.org/10.1016/j.reactfunctpolym.2005.05.011.
[3] A.M. Bezborodov, N.A. Zagustina, Enzymatic biocatalysis in chemical synthesis of
pharmaceuticals, Appl. Biochem. Microbiol. 52 (2016) 237–249, https://doi.org/
10.1134/S0003683816030030.
[29] R. Ullrich, M. Hofrichter, Enzymatic hydroxylation of aromatic compounds, Cell.
Mol. Life Sci. 64 (3) (2007) 271–293, https://doi.org/10.1007/s00018-007-6362-
1.
[30] F. Hollmann, I. Arends, D. Holtmann, Enzymatic reductions for the chemist, Green
Chem. 13 (2011) 2285–2314, https://doi.org/10.1039/c1gc15424a.
´
´
[31] F. Garzon-Posse, L. Becerra-Figueroa, J. Hernandez-Arias, D. Gamba-Sanchez,
Whole cells as biocatalysts in organic transformations, Molecules 23 (2018) 1265,
https://doi.org/10.3390/molecules23061265.
[4] F. Hollmann, D.J. Opperman, C.E. Paul, Biocatalytic reduction reactions from a
chemist’s perspective, Angew. Chem. Int. Ed. (2020), https://doi.org/10.1002/
anie.202001876.
[32] G.V. Bhasker Reddy, D.K. Joshi, M.H. Gold, Degradation of chlorophenoxyacetic
acids by the lignin-degrading fungus Dichomitus squalens, Microbiology 143 (7)
(1997) 2353–2360, https://doi.org/10.1099/00221287-143-7-2353.
[33] M. Horvat, S. Fritsche, R. Kourist, M. Winkler, Characterization of type IV
carboxylate reductases (CARs) for whole cell-mediated preparation of 3-
hydroxytyrosol, ChemCatChem 11 (16) (2019) 4171–4181, https://doi.org/
10.1002/cctc.v11.1610.1002/cctc.201900333.
[5] H.E. Schoemaker, D. Mink, M.G. Wubbolts, Dispelling the myths — biocatalysis in
industrial synthesis, Science 299 (2003) 1694–1697, https://doi.org/10.1126/
science.1079237.
[6] H. Sun, H. Zhang, E.L. Ang, H. Zhao, Biocatalysis for the synthesis of
pharmaceuticals and pharmaceutical intermediates, Bioorg. Med. Chem. 26 (7)
(2018) 1275–1284, https://doi.org/10.1016/j.bmc.2017.06.043.
[7] G.E. Silverman, The Organic Chemistry of Enzyme-Catalyzed Reactions, Academic
Press, San Diego, 2000.
[34] T.S. Zhuk, M. Goldmann, J. Hofmann, J.C.S. Pohl, H. Zorn, Preparative aerobic
oxidations with basidiomycetous enzymes: C-H-functionalization of adamantane,
J. Mol. Cat. B 122 122 (2015) 87–92, https://doi.org/10.1016/j.
molcatb.2015.08.002.
[8] A. Hage, H.E. Schoemaker, J.A. Field, Reduction of aryl acids by white-rot fungi for
the biocatalytic production of aryl aldehydes and alcohols, Appl. Microbiol.
Biotechnol. 52 (6) (1999) 834–838, https://doi.org/10.1007/s002530051600.
[9] G.P.A. Janusz, J. Sulej, U. Swiderska-Burek, A. Jarosz-Wilkołazka, A. Paszczynski,
Lignin degradation: microorganisms, enzymes involved, genomes analysis and
evolution, FEMS Microbiol. Rev. 41 (2017) 941–962, https://doi.org/10.1093/
femsre/fux049.
[35] C.E. Fabre, P.J. Blank, G. Goma, Production of benzaldehyde by several strains of
Ischnoderma benzoinum, Sci. Aliments 16 (1996) 61–68. ISSN 0240-8813.
[36] P. Baldrian, Purification and characterization of laccase from the white-rot fungus
Daedalea quercina and decolorization of synthetic dyes by the enzyme, Appl.
Microbiol. Biotechnol. 63 (5) (2004) 560–563, https://doi.org/10.1007/s00253-
003-1434-0.
[10] M. Madadi, A. Abbas, Lignin degradation by fungal pretreatment: a review, J. Plant
Pathol. Microbiol. 8 (2017) 1000398, https://doi.org/10.4172/2157-
7471.1000398.
[37] P. Giardina, G. Palmieri, B. Fontanella, V. Rivieccio, G. Sannia, Manganese
peroxidase isoenzymes produced by Pleurotus ostreatus grown on wood sawdust,
Arch. Biochem. Biophys. 376 (1) (2000) 171–179, https://doi.org/10.1006/
abbi.1999.1691.
[11] V.K. Ponnusamy, D.D. Nguyen, J. Dharmaraja, S. Shobana, J.R. Banu, R.
G. Saratale, S.W. Chang, G. Kumar, A review on lignin structure, pretreatments,
fermentation reactions and biorefinery potential, Bioresour. Technol. 271 (2019)
462–472, https://doi.org/10.1016/j.biortech.2018.09.070.
[38] N.-K. Krahe, R.G. Berger, F. Ersoy, A DyP-type peroxidase of Pleurotus sapidus with
alkene cleaving activity, Molecules 25 (2020) 1536, https://doi.org/10.3390/
molecules25071536.
[12] W. Wang, C. Zhang, X. Sun, S. Su, Q. Li, R.J. Linhardt, Efficient, environmentally-
friendly and specifc valorization of lignin: promising role of non-radical lignolytic
enzymes, World J. Microbiol. Biotechnol. 33 (2017) 125, https://doi.org/10.1007/
s11274-017-2286-6.
[39] A. Avram, A. Sengupta, P.H. Pfromm, H. Zorn, P. Lorenz, T. Schwarz, K.Q. Nguyen,
P. Czermak, Novel DyP from the basidiomycete Pleurotus sapidus: substrate
screening and kinetics, Biocatalysis 4 (2018) 1–13, https://doi.org/10.1515/boca-
2018-0001.
[13] R. Ullrich, J. Nuske, K. Scheibner, J. Spantzel, M. Hofrichter, Novel haloperoxidase
from the agaric basidiomycete Agrocybe aegerita oxidizes aryl alcohols and
aldehydes, Appl. Environ. Microbiol. 70 (2004) 4575–4581, https://doi.org/
10.1128/AEM.70.8.4575-4581.2004.
[40] J.E. Kowalczyk, M. Peng, M. Pawlowski, A. Lipzen, V. Ng, V. Singan, M. Wang, I.
¨
¨
V. Grigoriev, M.R. Makela, The white-rot basidiomycete Dichomitus squalens shows
highly specific transcriptional response to lignocellulose-related aromatic
compounds, Front. Bioeng. Biotechnol. 7 (2019), https://doi.org/10.3389/
fbioe.2019.00229.
[14] M. Hofrichter, R. Ullrich, Oxidation catalyzed by fungal peroxygenases, Curr. Opin.
Chem. Biol. 19 (2014) 116–125, https://doi.org/10.1016/j.cbpa.2014.01.015.
[15] C. Romero-Guido, A. Baez, E. Torres, Dioxygen activation by laccases: green
chemistry for fine chemical synthesis, Catalysts 8 (2018) 223, https://doi.org/
10.3390/catal8060223.
[41] A. Muheim, R. Waldner, M.S.A. Leisola, A. Fiechter, An extracellular aryl-alcohol
oxidase from the white-rot fungus Bjerkandera adusta, Enzyme Microb. Technol. 12
(3) (1990) 204–209, https://doi.org/10.1016/0141-0229(90)90039-S.
[42] H. Toogood, J. Gardiner, N. Scrutton, Biocatalytic reductions and chemical
versatility of the old yellow enzyme family of flavoprotein oxidoreductases,
ChemCatChem 2 (8) (2010) 892–914, https://doi.org/10.1002/cctc.201000094.
[43] J.S. Cha, H.C. Brown, Reaction of sodium aluminum-hydride with selected organic-
compounds containing representative functional-groups - comparison of the
[16] D.S. Arora, P.K. Gill, Effects of various media and supplements on laccase
production by some white rot fungi, Bioresour. Technol. 77 (1) (2001) 89–91,
https://doi.org/10.1016/S0960-8524(00)00114-0.
[17] D.W.S. Wong, Structure and action mechanism of ligninolytic enzymes, Appl.
Biochem. Biotechnol. 157 (2) (2009) 174–209, https://doi.org/10.1007/s12010-
008-8279-z.
7

T.S. Zhuk et al.
Bioorganic Chemistry 108 (2021) 104651
reducing characteristics of lithium and sodium aluminum hydrides, J. Org. Chem.
trichloro-1,3,5-triazine and PPh3 in the presence of K2CO3, RSC Adv. 4 (87) (2014)
46947–46950, https://doi.org/10.1039/C4RA08643K.
58 (17) (1993) 4727–4731, https://doi.org/10.1021/jo00069a042.
[44] G. Nagendra, C. Madhu, T.M. Vishwanatha, V.V. Sureshbabu, An expedient route
for the reduction of carboxylic acids to alcohols employing 1-propanephosphonic
acid cyclic anhydride as acid activator, Tetrahedron Lett. 53 (38) (2012)
5059–5063, https://doi.org/10.1016/j.tetlet.2012.06.108.
[46] X. Cui, Y. Li, C. Topf, K. Junge, M. Beller, Direct ruthenium-catalyzed
hydrogenation of carboxylic acids to alcohols, Angew. Chem. Int. Ed. 54 (36)
(2015) 10596–10599, https://doi.org/10.1002/anie.201503562.
[47] S.R. Joshi, K.L. Kataria, S.B. Sawant, J.B. Joshi, Kinetics of oxidation of benzyl
alcohol with dilute nitric acid, Ind. Eng. Chem. Res. 44 (2) (2005) 325–333,
https://doi.org/10.1021/ie0303911.
[45] S. Jaita, P. Kaewkum, C. Duangkamol, W. Phakhodee, M. Pattarawarapan, Solvent-
free reduction of carboxylic acids to alcohols with NaBH4 promoted by 2,4,6-
8