Role of laccase from Coriolus versicolor MTCC-138 in selective oxidation of aromatic methyl group

Article abstract of DOI:10.1134/S1068162014020034

Now a day, laccases are the most promising enzymes in the area of biotechnology and synthesis. One of the best applications of laccases is the selective oxidation of aromatic methyl group to aldehyde group. Such transformations are valuable because it is difficult to stop the reaction at aldehyde stage. Chemical methods used for such biotransformations are expensive and give poor yields. But, the laccase-catalyzed biotransformations of such type are non-expensive and yield is excellent. Authors have used crude laccase obtained from the liquid culture growth medium of fungal strain Coriolus versicolor MTCC-138 for the biotransformations of toluene, 3-nitrotoluene, and 4-chlorotoluene to benzaldehyde, 3-nitrobenzaldehyde, and 4-chlorobenzaldehyde, respectively, instead of purified laccase because purification process requires much time and cost. This communication reports that crude laccase can also be used in the place of purified laccase as effective biocatalyst.

Full text of DOI:10.1134/S1068162014020034

ISSN 1068ꢀ1620, Russian Journal of Bioorganic Chemistry, 2014, Vol. 40, No. 3, pp. 288–292. © Pleiades Publishing, Ltd., 2014.
Role of Laccase from Coriolus versicolor MTCCꢀ138
in Selective Oxidation of Aromatic Methyl Group1
Pankaj Kumar Chaurasia, Sunil Kumar Singh, and Shashi Lata Bharati2
Department of Chemistry, D.D.U. Gorakhpur University, Gorakhpur, Uttar Pradesh, 273009 India
Received August 30, 2013; in final form, September 23, 2013
Abstract—Now a day, laccases are the most promising enzymes in the area of biotechnology and synthesis.
One of the best applications of laccases is the selective oxidation of aromatic methyl group to aldehyde group.
Such transformations are valuable because it is difficult to stop the reaction at aldehyde stage. Chemical
methods used for such biotransformations are expensive and give poor yields. But, the laccaseꢀcatalyzed
biotransformations of such type are nonꢀexpensive and yield is excellent. Authors have used crude laccase
obtained from the liquid culture growth medium of fungal strain Coriolus versicolor MTCCꢀ138 for the
biotransformations of toluene, 3ꢀnitrotoluene, and 4ꢀchlorotoluene to benzaldehyde, 3ꢀnitrobenzaldehyde,
and 4ꢀchlorobenzaldehyde, respectively, instead of purified laccase because purification process requires
much time and cost. This communication reports that crude laccase can also be used in the place of purified
laccase as effective biocatalyst.
Keywords: laccase, Coriolus versicolor, mediator, HPLC, biotransformation
DOI: 10.1134/S1068162014020034
2 1
INTRODUCTION
However, it should not be generalized that only the
fungal system has ability to produce the laccases. Sevꢀ
eral reports can be referred to in the literature on proꢀ
duction of laccase in ascomycetes, such as Gaeumanꢀ
nomyces graminis [10], Magnaporthe grisea [11],
Ophiostoma novoꢀulmi [12], Mauginella [13], Melanoꢀ
carpus albomyces [14], Monocillium indicum [15],
Neurospora crassa [16], and Podospora anserine [17].
Laccase [EC 1.10.3.2] belongs to a group of
polyphenol oxidases containing copper atoms in the
catalytic center, which catalyze the reduction of
molecular oxygen to water [1–3]. It was first reported
in Japanese lacquer tree Rhus vernicifera [4]. Little is
known about higher plant laccases, probably due to
their presence in cell walls. Laccases are the lygnolytic In addition to plant pathogenic species, laccase proꢀ
enzymes and abundantly occur in the fungal systems duction was also reported for some soil ascomycete
species from the genera Aspergillus, Curvularia, and
Penicillium [18–20] and in some freshwater ascoꢀ
mycetes [21].
[5], mainly in ascomycetes, deuteromycetes, and
basidiomycetes; its production in lower fungi has never
been demonstrated. They occur in fungal causative
agents of the soft rot, in most, white rot causing fungi,
soil saprophytes, and edible fungi. These laccase proꢀ
ducing fungi are generally called wood degrading
fungi. White rot fungi are the highest producers of the
laccases, but litter decomposing and ectomicorrhizal
fungi also secrete laccases. Almost all white rot fungi
are laccase producers [5–7], except for Phanerochaete
chrysosporium. Some examples of the fungi which proꢀ
duce laccases are Pleurotus ostreatus, Coriolus sanꢀ
guineus, Trametes hirsuta, T. vercicolor, T. villosa,
Coriolopsis polyzona, Phlebia radiate, Podospora
anserine, Lentinus tigrinus, Pleurotus eryngii, Fomes
durrisimus MTCCꢀ1173, Pleurotus sajor caju
MTCCꢀ141, and Trametes trogii. Laccase is also
reported in bacteria, e.g. Azospirillum lipoferum [8], as
well as in wasp venom [9].
The ability of laccases to catalyze oxidation of varꢀ
ious phenolic, as well as nonꢀphenolic compounds,
coupled to the reduction of molecular oxygen to water
makes, it valuable from the point of view of their comꢀ
mercial applications [2, 22–24]. The importance of
laccases has increased after the discovery of mediators
[25, 26]. During the last two decades, laccases have
turned out to be the most promising enzymes for
industrial uses [23, 24] having applications in food,
pulps, paper, textile, cosmetics industries, and in synꢀ
thetic organic chemistry [27–30].
Selective oxidation of aromatic methyl group to its
aldehyde group has been done previously [31–33]
using purified laccase. Purification of enzyme is a long
process that needs highꢀcost chemical materials and
takes much time. In the present communication,
crude laccase obtained from liquid culture growth
medium containing natural lignin substrate, coir dust,
1
The article is published in the original.
2
Corresponding author: eꢀmail: shashilatachem@gmail.com.
288

ROLE OF LACCASE FROM Coriolus versicolor
Activity, IU/mL
289
0.40
0.35
0.30
0.25
0.20
0.15
0.10
0.05
6
0
2
4
10
12
14
Days
8
Fig. 1. Secretion of laccase by Coriolus versicolor MTCCꢀ138 in the liquid culture medium supplemented with different natural
lignin containing substrates saw dust ( ), corn cob ( ), wheat straw ( ), coir dust ( ), bagasse ( ), and control ( ).
×
᭿
᭡
᭜
᭺
of C. versicolor MTCCꢀ138 was used to demonstrate above, was used for selective biotransformation of aroꢀ
selective oxidation of aromatic methyl group in the matic methyl group to the corresponding aldehyde
presence of ABTS as mediator molecule. The purpose group.
of using crude laccase for such biotransformations was
to prove that the crude laccase is also an effective bioꢀ
catalyst like purified laccase and there is no need of
highꢀcost materials in order to obtain the crude lacꢀ
case. Thus, the novelty of this communication is in the
use of crude laccase obtained from C. versicolor
MTCCꢀ138, which was not reported so far, for such
selective oxidation reactions.
Selective transformations of aromatic methyl
group to its aldehyde group are, generally, a difficult
task in organic synthesis. The chemical routes of this
transformation are inconvenient because once a
methyl group is attacked, it is likely to be oxidized to
the carboxylic acid and it is very difficult to stop the
reaction at the aldehyde stage. Transformation of aroꢀ
matic methyl group to aldehyde group is one of the
best applications for laccases, who do it sharply within
1–3 h. The conversion done with crude laccase occurs
under milder conditions, the yield is >85%, and the
process is ecofriendly. The use of laccases for this purꢀ
pose has been studied in the presence of a mediator
molecule 2,2'ꢀazinoꢀbis(3ꢀethylbenzthiazolineꢀ6ꢀsulꢀ
fonic acid) diammonium salt (ABTS) [31]. Toluene,
3ꢀnitrotoluene, and 4ꢀchlorotoluene were successfully
converted to benzaldehyde, 3ꢀnitrobenzaldehyde, and
4ꢀchlorobenzaldehyde, respectively, in the presence of
ABTS as a mediator molecule.
RESULTS AND DISCUSSION
The experiment to define the maximum secretion
of the laccase in the liquid culture growth medium
amended with various lignin containing natural subꢀ
strates, like corn cob, coir dust, saw dust, wheat straw,
and bagasse particles, was performed with Coriolus
versicolor MTCCꢀ138. The control experiment had
similar medium composition except for the natural
lignin containing substrate being absent. The extracelꢀ
lular secretion of laccase was maximum in the case of
High performance liquid chromatography (HPLC).
the growth medium containing coir dust (Fig. 1). In In above enzyme catalyzed biotransformations, all
order to optimize secretion of laccase, it was studied in products formed are easily available and simple. So,
the presence of different amounts of coir dust. The authors have used HPLC technique to confirm the
maximum level of laccase was secreted in the liquid actual product formation by comparing the HPLC
culture medium containing 500 mg of the coir dust per chromatograms of standard aldehyde compounds with
25 mL of the culture medium (Fig. 2). The crude lacꢀ the enzymatically transformed compounds. The
case obtained from liquid culture growth medium with retention time of the standard sample of toluene was
a natural lignin substrate, coir dust, as described 4.21 min and the retention time of the standard sample
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PANKAJ KUMAR CHAURASIA et al.
Activity, IU/mL
0.50
0.45
0.40
0.35
0.30
0.25
0.20
0.15
0.10
0.05
6
8
0
2
4
10
12
Days
Fig. 2. Optimization of laccase secretion by Coriolus versicolor MTCCꢀ138 in liquid culture medium supplemented with different
amounts of coir dust: 100 mg ( ), 200 mg ( ), 400 mg ( ), 500 mg ( ), 600 mg ( ), 800 mg ( ), and 1000 mg (+).
᭜
ᮀ
᭡
×
᭿
of benzaldehyde was 3.52 min. The retention time of due to the aldehydic C–H bending. The results demꢀ
onstrate that the product was benzaldehyde.
the product of the enzymeꢀcatalyzed reaction
(3.50 min) coincided with the retention time of benꢀ
zaldehyde (3.52 min), confirming that the product of
enzyme catalyzed reaction was benzaldehyde. The
yield of the product was found to be 100%.
For expected product 3ꢀnitrobenzaldehyde, folꢀ
lowing spectral data were obtained. Band at
3010 cm–1 was due to the aromatic C–H stretching
while band at
C–H= 2900 cm–1 was due to the aldeꢀ
hydic C–H stretching. Conjugated aldehydic C=O
band appeared at
C=O = 1740 cm–1. Two bands obꢀ
tained at
N=O = 1500 cm–1, 1305 cm–1 were due to
νC–H =
ν
The similar type of reactions were done for 3ꢀnitroꢀ
toluene bioconversion to 3ꢀnitrobenzaldehyde and
4ꢀchlorotoluene bioconversion to 4ꢀchlorobenzaldeꢀ
hyde in the presence of ABTS as a mediator molecule.
The retention time of the standard samples of 3ꢀnitroꢀ
ν
ν
the asymmetric N=O stretching and symmetric N=O
stretching, respectively. A peak at 1390 cm–1 was due
toluene, 4ꢀchlorotoluene, 3ꢀnitrobenzaldehyde and to the aldehydic C–H bending.
4ꢀchlorobenzaldehyde were 6.58, 7.33, 5.91, and
For expected product 4ꢀchlorobenzaldehyde, IRꢀreꢀ
sults obtained are illustrated below. Appearance of a band
at
C–H = 3025 cm–1 was due to the C–H stretching
while appearance of a band at
C–H = 2830 cm–1 was
due to the aldehydic C–H stretching. Conjugated aldeꢀ
6.25 min, respectively. Thus, the retention time of the
products of the enzyme catalyzed reaction (5.96 and
6.23 min) revealed that the products of enzyme cataꢀ
lyzed reaction were 3ꢀnitrobenzaldehyde and 4ꢀchloꢀ
robenzaldehyde. In these cases, yield of the extracted
3ꢀnitrobenzaldehyde and 4ꢀchlorobenzaldehyde were
100 and 89%, respectively.
ν
ν
hydic C=O band appeared at ν
C=O = 1730 cm–1. A
peak at 1385 cm–1 was due to the aldehydic C–H
bending.
1H NMR (CDCl3, 300 MHz, ppm). 1H NMR specꢀ
tral data obtained for the expected product, i.e. benꢀ
Infrared (IR) spectroscopy. Identification and
characterization of products obtained during enzyꢀ
matic reaction were analyzed on the basis of IRꢀreꢀ
sults. IR results obtained for the expected benzaldeꢀ
zaldehyde, shows a singlet
the aldehydic proton, a doublet
due to ortho proton, a triplet = 7.36 ppm (2H) due
to meta proton and a triplet = 7.52 ppm (1H) due to
para proton.
δ
= 9.10 ppm (1H) due to
δ
= 7.83 ppm (2H)
hyde are the following: band at
was due to the aromatic C–H stretching, aldehydic
C–H stretching band came at
C–H = 2760 cm–1,
while conjugated aldehydic C=O stretching band
came at
ν
C–H = 3008 cm–1
δ
δ
ν
1H NMR spectral data obtained for the expected
ν
C=O = 1708 cm–1. A peak at 1345 cm–1 was product, i.e. 3ꢀnitrobenzaldehyde, shows a singlet
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ROLE OF LACCASE FROM Coriolus versicolor
291
δ
δ
= 9.56 ppm (1H) due to the aldehydic proton, a singlet media were inoculated with small pieces of mycelia
= 8.38 (1H) due to the proton located inꢀbetween (0.5 cm 0.5 cm) under aseptic condition and fungal
CHO and NO2 groups, a doublet at 8.12 (1H) due to cultures were grown under stationary culture condiꢀ
tions at 25 in a biological oxygen demand (BOD)
×
°C
the proton para to the CHO group, a triplet
δ = 7.95
incubator. In order to monitor the production of lacꢀ
case in the liquid culture medium, 0.5 mL aliquots of
the growth medium were withdrawn at regular interꢀ
vals of 24 hrs and filtered through sterilized Millipore
(1H) due to proton meta to the CHO group, and a
doublet 8.10 (1H) due to the proton ortho to the CHO
group.
1H NMR spectral data obtained for the expected
product, i.e. 4ꢀchlorobenzaldehyde, shows a singlet
filter 0.22 μm. The filtered extract was analyzed for the
activity of laccase using DMP as the substrate by the
method [35] given in assay section. Extracellular
secretion of laccase in the liquid culture medium by
C. versicolor MTCCꢀ138 was determined by plotting
the enzyme unit/mL of the growth medium against
the number of days after inoculation of the fungal
mycelia. In order to optimize the conditions for maxꢀ
imum production of laccase by C. versicolor
MTCCꢀ138 in the liquid culture medium, the amount
of the best inducer coir dust was varied from 100 mg to
1000 mg in 25 mL of the growth medium. The amount
of the inducer in the growth medium that gave the
maximum height of the enzyme activity peak was
taken as the optimal amount of the inducer.
δ
= 9.13 ppm (1H) for aldehydic proton, a doublet
7.87 (2H) ppm for protons ortho to the CHO group,
and a doublet 7.35 (2H) for the protons meta to the
CHO group.
In this way, all the above described results of
HPLC, IR and 1H NMR, collectively and clearly
demonstrate that the enzymatically transformed prodꢀ
ucts were benzaldehyde, 3ꢀnitrobenzaldehyde and
4ꢀchlorobenzaldehyde.
Thus, this communication reports the selective
oxidation of aromatic methyl group to aldehyde group
in the presence of ABTS with crude laccase obtained
from the fungal strain C. versicolor MTCCꢀ138 and
demonstrates that crude laccase is also an effective
biocatalyst, which can be used in the place of purified
laccase, successfully, for such oxidation reactions.
Enzyme assay. The assay solution 1.0 mL for DMP
as the substrate [35] contained 1.0 mM DMP in
50 mM sodium malonate buffer, pH 4.5, at 37
reaction was monitored by measuring the absorbance
= 468 nm and using the molar extinction
°C. The
change at
λ
EXPERIMENTAL
coefficient [35] value of 49.6 mM–1 cm–1. The UV/Vis
spectrophotometer Hitachi (Japan) model Uꢀ2900
fitted with electronic temperature control unit was
used for absorbance measurement. The least count of
absorbance measurement was 0.001 absorbance unit.
Materials. 4ꢀChlorotoluene was from Sigma
Chemical Company, St. Louis (USA). 2,2'ꢀAzinoꢀ
bis(3ꢀethylbenzthiazolineꢀ6ꢀsulphonic acid) diamꢀ
monium salt (ABTS) and 2,6ꢀdimethoxyphenol
(DMP) were from Fluka, Chemi New Ulm (Switzerꢀ
land). All other chemicals used in these investigations
were either from Himedia laboratory Ltd. Mumbai
(India) or from E. Merck Ltd. Mumbai (India) and
were used without further purifications.
Fungal strain and its growth. The fungal strain was
procured from the Microbial Type Culture Collection
Center and Gene Bank, Institute of Microbial Techꢀ
nology, Chandigarh, India, and was maintained on
agar slant as reported in MTCC Catalogue of strainsꢀ
2000 [34]. The growth medium for the fungal strain
Coriolus versicolor MTCCꢀ138 consisted of yeast
extract, 5.0 g, glucose, 10.0 g, and agar, 15.0 g, in 1.0 L
MilliꢀQ water.
One enzyme unit produced 1 μmol of the product per
minute under the specified assay conditions.
Biotransformations in the presence of ABTS as meꢀ
diator. The biotransformation of toluene to benzaldeꢀ
hyde [31–33] was performed in 14 mL of 100 mM soꢀ
dium acetate buffer, pH 4.5, containing 20 mM toluꢀ
ene in 30 mL of dioxane, 0.1 mM ABTS and 500 μL of
crude laccase kept in a 250 mL conical flask which was
stirred vigorously for 30 minutes (completion of the reacꢀ
tion was confirmed by spectrophotometry measurement
on the UV/Vis spectrophotometer Hitachi (Japan) modꢀ
el Uꢀ2900). The reaction solution was extracted twice
with 40 mL of ethyl acetate. Aliquot of 20
acetate extract was injected in Waters HPLC Model 600E
using spherisorb C18column (5 UV, 4.5 250 mm). The
mobile phase was consisted methanol, flow rate was
0.5 mL/min. The detection was performed using a Waꢀ
ters UV detector model 2487 at = 254 nm.
μL of the ethyl
×
In order to detect the extracellular secretion of lacꢀ
case by Coriolus versicolor MTCCꢀ138, liquid culture
growth medium reported by Coll et al. [35] was used.
This medium consisted of glucose, 10.0 g, asparagines,
λ
Biotransformations of 3ꢀnitrotoluene to 3ꢀnitroꢀ
benzaldehyde and 4ꢀchlorotoluene to 4ꢀchlorobenꢀ
zaldehyde were also studied using the same method as
described above, except for the fact that times of stirꢀ
ring of the reaction solutions were one and two hours,
respectively, in these cases.
1.0 g, yeast extract, 0.5 g, MgSO4
⋅ 7H2O, and
FeSO4 7H2O, 0.01 g in 1.0 L of MilliꢀQ water. The
⋅
above liquid culture growth medium containing natuꢀ
ral lignin substrates like coir dust, corn cob, wheat
straw, saw dust, and bagasse particles were separately
prepared by adding 0.5 g of one of the natural lignin
substrates to 25 mL of growth medium in 100 mL culꢀ
All the synthesized products were extracted from
ture flasks which were sterilized. The sterilized growth their reaction solutions with ethyl acetate and these
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PANKAJ KUMAR CHAURASIA et al.
15. Thakker, G.D., Evans, C.S. and Rao, K.K., Appl.
Microbiol. Biotechnol., 1992, vol. 37, no. 3, pp. 321–
323.
extracted products were identified and characterized
by HPLC, IR, and 1H NMR techniques. Since yields
of the products were 100% except 4ꢀchlorobenzaldeꢀ
hyde, where yield was 89%, we did not purify the prodꢀ
ucts any further.
16. Froehnerand, S.C. and Eriksson, K.E., J. Bacteriol.
,
,
,
1974, vol. 120, no. 1, pp. 458–465.
17. Molitoris, H.P. and Esser, K., Archiv. für. Mikrobiologie
1970, vol. 72, no. 3, pp. 267–296.
ACKNOWLEDGMENTS
18. Banerjee, U.C. and Vohra, R.M., Folia Microbiologica
1999, vol. 36, no. 4, pp. 343– 346.
The authors acknowledge the financial support of
CSIRꢀHRDG, New Delhi for the award of JRF and
SRF (NET), award no. 09/057(0201)/2010ꢀEMRꢀI
to Mr. Pankaj Kumar Chaurasia. Dr. S.K Singh is also
19. Rodr
de la Fuente, G., and Trojanowski, J., Appl. Microbiol.
Biotechnol., 1996, vol. 45, no. 3, pp. 399–403
ιgꢁ uez, A., Falcoꢁn, M.A., Carnicero, A., Perestelo, F.,
.
thankful to CSIRꢀHRDG, New Delhi, for the award 20. Scherer, M. and Fischer, R., Archiv. Microbiol, 1998,
vol. 170, no. 2, pp. 78–84.
21. AbdelꢀRaheem, A. and Shearer, C.A., Fungal Diversity
2002, vol. 11, pp. 1–19.
22. Wandrey, C., Liese, A. and Kihumbu, D., Org. Proc.
Res. Develop., 2000, vol. 4, pp. 285–290.
of RA, award no. 09/057(0206)/2012ꢀEMRꢀI. The
authors are thankful to Prof. K.D.S. Yadav, Emeritus
Scientist, CSIR, New Delhi, for his helpful discusꢀ
sions.
,
23. Couto, S.R. and Harrera, J.L.T., Biotechnol. Advan.
,
REFERENCES
2006, vol. 24, no. 500–513.
1. Riva, S., Trends Biotechnol., 2006, vol. 24, no. 5,
24. Xu, F., Ind. Biotechnol., 2005, vol. 1, pp. 38–50.
25. Acunzo, D.F. and Galli, C., J. Europ. Biochem., 2003,
pp. 219–226.
2. Baldrian, P., FEMS Microbiol. Rev., 2006, vol. 30,
vol. 270, pp. 3634–3640.
pp. 215–242.
26. Morozova, O.V., Shumakovich, G.P., Shleev, S.V. and
Yaropolov, Y.I., Biochem. Microbiol., 2003, vol. 43,
pp. 523–535.
27. Coniglio, A., Galli, C. and Gentili, P., J. Mol. Cat. B:
Enzymatic, 2008, vol. 50, no. 1, pp. 40–49.
28. Mikolasch, A., Niedermeyer, T.H.J.. Lalk, M., Witt, S.,
Seefeld, S., Hammer, E., Schauer, F., Gesell, M., Hesꢀ
sel, S., Julich, W.D. and Lindoquist, D.U., Chem.
Pharma. Bullet., 2006, vol. 54, no. 5, pp. 632–638.
3. Dwivedi, U.N., Singh, P., Pandey, V.P., and Kumar, A.,
J. Mol. Cat. B: Enzymatic, 2011, vol. 68, no. 2, pp. 117–
128.
4. Yoshida, H., J. Chem. Soc., 1883, vol. 43, pp. 472–486.
5. Messerschmidt, A., MultiꢀCopper Oxidases, Singapore:
World Scientific, 1997.
6. Mayer, A.M. and Staples, R.C., Phytochemistry, 2002,
vol. 60, pp. 551–565.
7. Bertrand, G., C. R. Heabd. Seances Acad. Sci., 1896,
vol. 123, pp. 463–464.
8. Givaudan et al., FEMS Micribiol. Lett., 1993, vol. 108,
29. Mikolasch, A., Niedermeyer, T.H.J., Lalk, M., Witt, S.,
Seefeld, S., Hammer, E., Schauer, F., Salazar, G., Hesꢀ
sel, S., Julich, W.D. and Lindequist, U., Chem.
Pharma. Bullet., 2007, vol. 55, no. 3, pp. 412–416.
pp. 205–210.
30. Mikolasch, A., Hammer, E., Jonas, U., Popowski, K.,
Stielow, A. and Schaner, F., Tetrahedron, 2002, vol. 58,
pp. 7589–7593.
9. Perkinson, N., Smith, I., Weave, R. and Edwards, J.P.,
Insect. Biochem. Mol. Biol., 2001, vol. 31, pp. 57–63.
10. Edens, W.A., Goins, T.Q., Dooley, D., and Henson, J.M.,
Appl. Environ. Microbiol., 1999, vol. 65, no. 7,
pp. 3071–3074.
31. Potthest, A., Rosenanu, T., Chen, C.ꢀL., and Gratzl, J.S.,
J. Org. Chem., 1995, vol. 60, pp. 4320–4321.
32. FritzꢀLanghals Kunath, B., Tetrahedron Lett., 1998,
11. Iyer, G. and Chattoo, B.B., FEMS Microbiol. Lett.
2003, vol. 227, no. 1, pp. 121–126.
,
vol. 39, pp. 5955–5956.
33. Sahay, R., Yadav, R.S.S., and Yadava, S., Appl. Biochem.
Biotechnol., 2012, vol. 166, pp. 563–575.
12. Binz, T. and Canevascini, G., Curr. Microbiol., 1997,
vol. 35, no. 5, pp. 278–281.
34. Catalogue of Strainsꢀ2000, 5th ed., Chandigarh: Microꢀ
bial Type Culture Collection and Gene Bank Institute
of Microbial Technology.
13. Palonen, H., Saloheimo, M., Viikari, L. and Kruus, K.,
Enz. Microb. Technol., 2003, vol. 33, no. 6, pp. 854–
862.
14. Kiiskinen, L.L., Viikari, L., and Kruus, K., Appl. 35. Coll, M.P., FernandezꢀAbalos, J.M., Villomueva, J.R.,
Microbiol. Biotechnol., 2002, vol. 59, no. 2–3, pp. 198–
204.
Somtamariaa, R., and Perez, P., Appl. Environ. Microꢀ
biol., 1993, vol. 59, pp. 2607–2613.
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 40
No. 3
2014