Previous Article | Next Article 
Applied and Environmental Microbiology, June 2000, p. 2290-2296, Vol. 66, No. 6
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Distribution of Aldoxime Dehydratase in
Microorganisms
Yasuo
Kato,
Ryoko
Ooi, and
Yasuhisa
Asano*
Biotechnology Research Center, Faculty of
Engineering, Toyama Prefectural University, Kosugi, Toyama
939-0398, Japan
Received 20 December 1999/Accepted 21 March 2000
 |
ABSTRACT |
The distribution of phenylacetaldoxime-degrading and
pyridine-3-aldoxime-degrading ability was examined with intact cells of
975 microorganisms, including 45 genera of bacteria, 11 genera of
actinomyces, 22 genera of yeasts, and 37 genera of fungi, by monitoring
the decrease of the aldoximes by high-pressure liquid chromatography.
The abilities were found to be widely distributed in bacteria,
actinomyces, fungi, and some yeasts: 98 and 107 strains degraded
phenylacetaldoxime and pyridine-3-aldoxime, respectively. All of the
active strains exhibited not only the aldoxime-dehydration activity to
form nitrile but also nitrile-hydrolyzing activity. On the other hand,
all of 19 nitrile-degrading microorganisms (13 species, 7 genera) were
found to exhibit aldoxime dehydration activity. It is shown that
aldoxime dehydratase and nitrile-hydrolyzing activities are widely
distributed among 188 aldoxime and 19 nitrile degraders and that the
enzymes were induced by aldoximes or nitriles.
| |
INTRODUCTION |
Nitrile compounds are discharged
into the environment as industrial waste water, agricultural chemicals,
etc. (23). Many microorganisms can use nitriles as a source
of carbon and/or nitrogen for growth. Asano et al. have isolated
various nitrile-degrading microorganisms from soil (2, 5,
24) and clarified that nitriles are converted to carboxylic acids
either by a combination of nitrile hydratase (1, 3, 6) and
amidase (4) or by nitrilase (8). These enzymes
have been extensively evaluated from the viewpoint of chemical
industry: the industrial production of acrylamide, nicotinamide, and
5-cyanovalelamide are typical examples (5, 6, 16, 26).
Despite the importance of the nitrile-hydrolyzing enzymes in industry,
information about their distribution in microorganisms is quite
limited, and their physiological role has never been well understood
since they have been screened only from nitrile-degrading microorganisms.
We have been studying aldoxime-degrading enzymes and isolated various
aldoxime-degrading microorganisms, e.g., Bacillus sp. strain
OxB-1 (7) and Rhodococcus sp. strain YH3-3
(12), from soil. The isolated strains metabolized aldoximes
through nitriles into the corresponding carboxylic acid by a
combination of a novel aldoxime dehydratase and nitrile-hydrolyzing
enzymes (7, 12) (Fig. 1). The
novel aldoxime dehydratase was purified and characterized from
Bacillus sp. strain OxB-1 (7, 15). The enzyme
from Rhodococcus sp. strain YH3-3 was applied to the
enzymatic synthesis of nitriles from aldoximes under a mild condition
(12, 13). A nitrile hydratase responsible for aldoxime
metabolism was purified and characterized from Rhodococcus
sp. strain YH3-3, and its properties were compared with the known
nitrile hydratases (14).
View larger version (7K):
[in this window]
[in a new window]
|
FIG. 1.
Microbial metabolism of aldoximes. Bacillus
sp. strain OxB-1 metabolizes PAOx (R = PhCH2) to form
PAN, which is successively hydrolyzed to PAA by the action of nitrilase
(7). On the other hand, Rhodococcus sp. strain
YH3-3 metabolizes PyOx (R = 3-pyridyl) as follows: the aldoxime is
dehydrated to form CyPy, which is converted to NAm by nitrile
hydratase, and the NAm is successively hydrolyzed to NA by amidase
(12).
|
|
To elucidate the generality of the relationship of aldoxime dehydratase
and nitrile-hydrolyzing enzymes in microorganisms, we examined the
distribution of both enzyme activities with the intact cells of a
variety of aldoxime- or nitrile-degrading microorganisms, monitoring
the decrease of the aldoximes, and we studied the relationship among
the enzymes.
| |
MATERIALS AND METHODS |
Chemicals.
Meat and malt extracts were obtained from Kyokuto
(Tokyo, Japan). Polypepton and yeast extract were purchased from Nippon Seiyaku (Tokyo, Japan). High-pressure liquid chromatography (HPLC) columns ODS-80Ts (4.6 by 150 mm) and Hibar LiChrosorb-NH2
(4.0 by 250 mm) were from Tosoh Corp. (Tokyo, Japan) and Kanto
Chemicals (Tokyo, Japan), respectively. Aldoximes were synthesized as
described previously (7, 12). All other chemicals were from
commercial sources and used without further purification.
Microorganisms and culture media.
A total of 975 strains
from the following type culture collections were used: the Institute of
Molecular and Cellular Biosciences (IAM), University of Tokyo, Tokyo,
Japan; the Institute of Fermentation (IFO), Osaka, Japan; the Japan
Collection of Microorganisms (JCM), Tokyo, Japan; the National
Collections of Industrial Food and Marine Bacteria (NCIMB), Aberdeen,
Scotland; the National Collections of Type Cultures and Pathogenic
Fungi (NCTC) London, United Kingdom; the American Type Culture
Collection (ATCC); and our own laboratory (TPU). These included 407 strains of 45 genera of bacteria, 133 strains of 11 genera of
actinomyces, 333 strains of 22 genera of yeasts, and 102 strains of 37 genera of fungi.
The culture medium for the bacteria was composed of 1.0% of meat
extract, 1.0% of Polypepton, and 0.5% NaCl (pH 7.2). The
medium for
actinomyces contained 1.0% malt extract, 0.4% yeast
extract, and
0.4%
D-glucose (pH 7.2). The medium for yeasts consisted
of 1.0%
D-glucose, 0.5% Polypepton, 0.3% yeast extract,
and 0.3%
malt extract (pH 5.6). The medium for fungi contained 20 g of
sucrose and boiled extract from 200 g of potato per liter (pH
6.0). Tap water was used for the media described above, and the
pH was
adjusted by using HCl and NaOH. Each microorganism was
inoculated into
a test tube containing 10 ml of the medium with
0.05% concentrations
of the inducers, i.e., phenylacetaldoxime
(PAOx), pyridine-3-aldoxime
(PyOx), phenylacetonitrile (PAN),
and 3-cyanopyridine (CyPy), and then
incubated with shaking for
2 to 14 days at 30°C until their growth
reached maximum levels.
For facultative anaerobic bacteria, the strains
were also grown
under static
conditions.
Enzyme assay and definition of units.
Microbial degradation
of aldoxime was qualitatively analyzed by thin-layer chromatography
(TLC). After the strain was cultured in the medium containing PAOx or
PyOx, 1 µl of the culture was spotted onto silica gel TLC plates
(Kieselgel 60; Merck) and then developed with 20% (vol/vol) ethyl
acetate in hexane. The remaining aldoxime in the medium was visualized
with iodine vapor.
Aldoxime dehydration and nitrile-hydrolyzing activities were
quantitatively assayed by measuring the rate of consumption of
aldoxime
and nitrile, respectively. A standard assay solution
contained 50 µmol of potassium phosphate buffer (pH 7.0), 25 µmol
of substrate,
and washed cells from 5 ml of culture in a total
volume of 500 µl.
After an incubation at 30°C with shaking, the
reaction mixture was
centrifuged (18,000 ×
g, 2 min) at 4°C to
remove the
cells. The supernatant was analyzed with a Waters 600E
HPLC apparatus
equipped with a Waters 486 absorbance detector
(254 nm) at a flow rate
of 1.0 ml/min. PAOx, PAN, phenylacetamide
(PAAm), and phenylacetic acid
(PAA) were detected with an ODS-80Ts
column using an elution solvent
consisting of 10 mM H
3PO
4 in 40%
(vol/vol)
CH
3CN, and PyOx and 3-CyPy were measured with the same
column using 10 mM H
3PO
4 in 10% (vol/vol)
CH
3CN. Nicotinamide
(NAm) and nicotinic acid (NA) were
detected with Hibar LiChrosorb-NH
2 column with 2.5 mM
potassium phosphate buffer (pH 2.8) in 75%
(vol/vol)
CH
3CN. Single units of aldoxime dehydration and
nitrile-hydrolyzing
activities were defined as the amount of enzyme
that catalyzed
the dehydration of aldoxime and hydrolysis of nitrile,
respectively,
at a rate of 1 µmol/min. Reaction mixtures with the
cells but
without substrates served as blanks for both the assays.
Under
the assay conditions, the measurable detection limit of the
enzyme
activity was about 0.01 (in units/liter of
culture).
| |
RESULTS |
Distribution of PAOx and PyOx dehydration activity in bacteria and
actinomyces.
We selected PAOx and PyOx as a model compound of
alkylaldoxime and arylaldoxime, respectively. Bacterial and
actinomycetal strains in our stock cultures were aerobically grown in
the medium containing PAOx or PyOx, and the ability to degrade the
aldoximes was screened by qualitative measurement of aldoxime
disappearance in the culture medium by using TLC assay. Among the 540 strains tested, 37 strains were able to degrade PAOx, and this activity was distributed across 7 genera, while 96 strains, distributed across
30 genera, could degrade PyOx. Although some aerobically grown
facultative anaerobes could degrade PyOx, none of the statically grown
cells could degrade PAOx nor PyOx. We measured not only the aldoxime
dehydration activities but also the PAN- and CyPy-hydrolyzing activities in the cells of PAOx- or PyOx-degrading strains. Since all
of the aldoxime-degrading strains contained both aldoxime dehydratase
and nitrile-hydrolyzing enzymes, nitriles formed by the dehydration of
aldoximes were subsequently converted to amides and/or acids by the
action of the latter enzymes. We therefore analyzed the dehydration
activity by measuring the consumption of aldoxime in the assay.
Nitrile-hydrolyzing activity was estimated by measuring the rate of
consumption of nitrile, which was equal to the sum of nitrile hydratase
and nitrilase activities. As shown in Tables
1 and 2,
all of the strains grown with PAOx or PyOx exhibited the aldoxime
dehydration activities, together with nitrile-hydrolyzing enzyme
activities. The 21 strains marked with a number symbol (#) exhibited
both PAOx and PyOx dehydration activities.
By measuring the formation of PAAm and NAm in the reaction mixture, we
determined the mode of nitrile degradation by the active
strains.
Totals of 10 and 21 strains, marked with an asterisk
(*) in Tables
1
and
2, respectively, converted nitrile to acid-accumulating
amide,
indicating that the strains hydrolyzed the nitriles by
a sequential
action of nitrile hydratase and amidase. These results
do not exclude
the possibility of a coexistence of nitrilase in
the strains. The other
strains degraded PAN and CyPy without forming
the corresponding amides.
In these strains, the nitriles are hydrolyzed
by nitrilase and/or the
combination of nitrile hydratase and amidase
in which amidase activity
was stronger than that of nitrile
hydratase.
Distribution of aldoxime dehydration activity in fungi.
We
next screened for fungal strains which degrade PAOx or PyOx.
PAOx-degrading activity was widely distributed in various genera. Of
the 102 strains, 52 among 28 genera were able to degrade PAOx, whereas
PyOx-degrading activity was seen in only a limited number of fungal
strains; it was seen in 11 strains among 3 genera. Tables
3 and 4
show both of the enzyme activities in all of the active strains. Some
active fungal strains degraded PAN accumulating PAAm, suggesting that
the strains hydrolyzed PAN by the combinatorial action of nitrile
hydratase and amidase, whereas PyOx-degrading fungi converted CyPy into
NA without forming NAm.
Distribution of aldoxime dehydration activity in yeasts.
Aldoxime degradation activity in yeasts is rare. We screened 333 strains, including 2 species of Brettanomyces, 29 species of
Candida, 3 species of Cryptococcus, 3 species of
Debaryomyces, 1 species of Endomycopsis, 1 species of Hanseniaspora, 24 species of Hanenula,
3 species of Kloeckera, 1 species of Lipomyces,
19 species of Pichia, 3 species of
Rhodosporidium, 10 species of Rhodotorula, 32 species of Saccharomyces, 1 species of
Saccharomycodes, 1 species of Saccharomycopsis, 3 species of Schizosaccharomyces, 1 species of
Sporidiobolus, 4 species of Sporobolomyces, 12 species of Torulopsis, 3 species of Trichosporon,
and 1 species of Zygosaccharomyces. Of these, two were
active in degrading PAOx: Candida methanolica TPU 1217 and
Pichia miso TPU 1306 showed PAOx dehydration activity at
1.31 and 0.9 (U/liter of culture), coexisting with a PAN-hydrolyzing activity at 0.5 and 0.2 (U/liter of culture), respectively, when they
were grown with 0.05% of PAOx. It was also shown that both the strains
degraded PAOx by a successive combination of aldoxime dehydration and
nitrile-hydrolyzing enzymes. P. miso degraded PAN
accumulating PAAm, while C. methanolica degraded it without the formation of PAAm. On the other hand, no yeast strain could degrade PyOx.
Based on these results, it is shown that all of the PAOx or PyOx
degraders degraded the aldoximes by a combination of aldoxime
dehydratase and nitrile-hydrolyzing
enzymes.
Aldoxime dehydration enzyme activity in nitrile-degrading
microorganisms.
Nineteen microorganisms (13 species, 7 genera)
which had been isolated as nitrile degraders were grown with aldoximes,
such as PAOx and PyOx, or nitriles, such as PAN and CyPy, and the
aldoxime dehydration and nitrile-hydrolyzing activities were measured. As shown in Table 5, all of the nitrile
degraders thus examined showed either PAOx or PyOx dehydratase
activities. A high PAOx dehydration activity was seen in
alkylnitrile-degrader, e.g., Rhodococcus sp. strain N-774
(23), Corynebacterium sp. strain C-5
(22), R. rhodochrous J-1 (3, 19), and
R. erythropolis BG-16 (Y. Asano, T. Yasuda, Y. Tani, and H. Yamada, Abstr. Ann. Meet. Japan Soc. Ferment. Technol., 1981). No
aldoxime dehydratase activity was seen when the strains were grown
without aldoximes or nitriles, although PAN- or CyPy-hydrolyzing
activity was detected in the cells.
View this table:
[in this window]
[in a new window]
|
TABLE 5.
Aldoxime dehydration and nitrile-hydrolyzing activities
in nitrile-degrading microorganisms when they were grown with or
without inducersa
|
|
| |
DISCUSSION |
Aldoximes are considered to be intermediates in the
biosynthesis of certain biologically active compounds such as
indoleacetic acid, cyanogenic glucosides, and glucosinolates in plants
(17, 20, 21); however, very little is known about
aldoxime-degrading enzymes: indoleacetaldoxime (IAOx) hydrolyase (EC
4.2.1.29), catalyzing a specific dehydration reaction of IAOx to form
indoleacetonitrile, has been detected in limited genera of fungi and
higher plants (20, 21), and the enzyme has only been
partially purified from Gibberella spp. (17). In
our previous report, we isolated various aldoxime-degrading bacteria
from soil by several-months' acclimation and clarified that the
isolated strains degraded aldoximes by the combination of aldoxime
dehydratase and nitrile-hydrolyzing enzymes (7, 12).
Rhodococcus sp. strain YH3-3, isolated in a medium
containing PyOx as a sole source of nitrogen, degrades PyOx to NA by a
combination of aldoxime dehydratase, nitrile hydratase, and amidase
(12) (Fig. 1), whereas Bacillus sp. strain OxB-1 degrades PAOx to PAA by the action of aldoxime dehydratase and nitrilase (7) (Fig. 1), although it could not utilize PAOx as a nitrogen or carbon source due to the toxicity of PAOx. In this
report, we discovered that the novel aldoxime dehydratases acting on
the aldoximes are distributed among a variety of microorganisms, together with nitrile-hydrolyzing enzymes. We also screened for the
aldoxime dehydratase activity from the nitrile degraders hitherto known
and clarified that all of the nitrile degraders contained not only
nitrile-hydrolyzing enzymes but also aldoxime dehydratase by culturing
with aldoxime. None of the active strains showed aldoxime
dehydration activity when they were grown in the medium without
aldoximes, suggesting that aldoxime dehydratase was inducibly formed in
these strains. Although a number of studies had been done on the
nitrile-hydrolyzing enzymes, their induction mechanisms remained
unknown. Nitrile hydratases of P. chlororaphis B23 and R. rhodochrous J-1 are strongly induced by amides and their
derivatives (26), although that of Rhodococcus
sp. strain N-774 (23) is formed constitutively. Nitrilases
are inducibly formed (2, 9, 10, 18, 24, 25, 27), except that
the enzyme is constitutively produced in Acinetobacter sp.
strain AK 226 (27). In the present study we found that
aldoximes act as good inducers not only for aldoxime dehydratase but
also for the nitrile-hydrolyzing enzymes. From the results presented
here, we cannot clarify whether aldoxime dehydratase and
nitrile-hydrolyzing enzymes are regulated separately or not in these
strains. We have recently discovered that the genes for aldoxime
dehydratase and nitrilase coexisted as a gene cluster in the genome of
Bacillus sp. strain OxB-1 (15). It is important
to genetically clarify the general relationship between aldoxime
dehydratase and nitrile-hydrolyzing enzymes in the aldoxime- and
nitrile-degrading strains.
A total of 31 bacterial and 9 fungal aldoxime and nitrile degraders
showed both the PAOx and PyOx dehydration activities, together with PAN
and CyPy hydrolyzing activities. It is not clear whether the
differences between PAOx and PyOx dehydration activities observed in
the strains were simply due to the substrate specificity of aldoxime
dehydratase or to the possible existence of two or more aldoxime
dehydratases having different substrate specificities. Purification and
characterization of aldoxime dehydratase and nitrile-hydrolyzing
enzymes from the strains will solve this problem.
It has been reported that about 70 strains of microorganisms were
isolated to degrade nitrile compounds, and they were distributed across
37 species of 19 genera (6, 24). However, they appear frequently in limited genera, i.e., Arthrobacter,
Bacillus, Rhodococcus, Fusarium, since
the most common screening protocols involve enrichment isolation in the
media containing nitriles as a carbon or a nitrogen source: this is
inevitably skewing positive clones to groups of readily culturable
microorganisms. By culturing the strains with aldoxime, we showed here
that the occurrence of nitrile hydratase in various species of
facultative anaerobes and fungi. Although it is reported that
Rhodococcus sp. strain AK32 has only nitrilase (27), culturing the strain with PAOx showed that the strain also contains nitrile hydratase and amidase in addition to nitrilase. These findings encourage us to screen further cryptic
nitrile-hydrolyzing enzymes in microorganisms by culturing with aldoxime.
| |
ACKNOWLEDGMENTS |
This work was supported in part by a Grant-in-Aid for "Research
for the Future" (JSPS-RFTF 96I00302) from the Japan Society for the
Promotion of Science.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: Biotechnology
Research Center, Faculty of Engineering, Toyama Prefectural University, Kosugi, Toyama 939-0398, Japan. Phone: 81-766-56-7500, ext. 530. Fax:
81-766-56-2498. E-mail: asano{at}pu-toyama.ac.jp.
| |
REFERENCES |
| 1.
|
Asano, Y.,
Y. Tani, and H. Yamada.
1980.
A new enzyme "nitrile hydratase" which degrades acetonitrile in combination with amidase.
Agric. Biol. Chem.
44:2251-2252.
|
| 2.
|
Asano, Y.,
S. Ando,
Y. Tani,
H. Yamada, and T. Ueno.
1981.
Fungal degradation of triacrylonitrile.
Agric. Biol. Chem.
45:57-62.
|
| 3.
|
Asano, Y.,
K. Fujishiro,
Y. Tani, and H. Yamada.
1982.
Aliphatic nitrile hydratase from Arthrobacter sp. J-1. Purification and characterization.
Agric. Biol. Chem.
46:1165-1174.
|
| 4.
|
Asano, Y.,
M. Tachibana,
Y. Tani, and H. Yamada.
1982.
Purification and characterization of amidase which participates in nitrile degradation.
Agric. Biol. Chem.
46:1175-1181.
|
| 5.
|
Asano, Y.,
T. Yasuda,
Y. Tani, and H. Yamada.
1982.
A new enzymatic method of acrylamide production.
Agric. Biol. Chem.
46:1183-1189.
|
| 6.
|
Asano, Y.
1991.
Studies on the synthesis of amides and amino acids by a novel microbial enzymes.
Nippon Nogeikagaku Kaishi
65:1617-1626.
|
| 7.
|
Asano, Y., and Y. Kato.
1998.
Z-Phenylacetaldoxime degradation by a novel aldoxime dehydratase from Bacillus sp. strain OxB-1.
FEMS Microbiol. Lett.
158:185-190[CrossRef].
|
| 8.
|
Bandyopadhyay, A. K.,
T. Nagasawa,
Y. Asano,
K. Fujishiro,
Y. Tani, and H. Yamada.
1986.
Purification and characterization of benzonitrilases from Arthrobacter sp. strain J-1.
Appl. Environ. Microbiol.
51:302-306[Abstract/Free Full Text].
|
| 9.
|
Harper, D. B.
1976.
Purification and properties of an unusual nitrilase from Nocardia NCIMB 11216.
Biochem. Soc. Trans.
4:502-504[Medline].
|
| 10.
|
Harper, D. B.
1985.
Characterization of a nitrilase from Nocardia sp. (rhodochrous group) NCIMB 11215, using p-hydroxybenzonitrile as sole carbon source.
Int. J. Biochem.
17:677-683[CrossRef][Medline].
|
| 11.
|
Kakeya, H.,
N. Sakai,
T. Sugai, and H. Ohta.
1991.
Microbial hydrolysis as a potent method for the preparation of optically active nitriles, amides, and carboxylic acids.
Tetrahedron Lett.
32:1343-1346[CrossRef].
|
| 12.
|
Kato, Y.,
R. Ooi, and Y. Asano.
1998.
Isolation and characterization of a bacterium possessing a novel aldoxime-dehydration activity and nitrile-degrading enzymes.
Arch. Microbiol.
170:85-90[CrossRef][Medline].
|
| 13.
|
Kato, Y.,
R. Ooi, and Y. Asano.
1999.
A new enzymatic method of nitrile synthesis by Rhodococcus sp. strain YH3-3.
J. Mol. Catal. B Enzymatic
6:249-256.
|
| 14.
|
Kato, Y.,
T. Tsuda, and Y. Asano.
1999.
Nitrile hydratase involved in aldoxime metabolism from Rhodococcus sp. strain YH3-3 purification and characterization.
Eur. J. Biochem.
263:662-670[Medline].
|
| 15.
|
Kato, Y.,
K. Nakamura,
H. Sakiyama,
S. G. Mayhew, and Y. Asano.
2000.
A novel heme-containing lyase, phenylacetaldoxime dehydratase from Bacillus sp. strain OxB-1: purification, characterization, and molecular cloning of the gene.
Biochemistry
39:800-809[CrossRef][Medline].
|
| 16.
|
Kobayashi, M.,
T. Nagasawa, and H. Yamada.
1992.
Enzymatic synthesis of acrylamide: a success story not yet over.
Trends Biotechnol.
10:402-408[CrossRef][Medline].
|
| 17.
|
Mahadevan, S.
1973.
Role of oximes in nitrogen metabolism in plants.
Annu. Rev. Plant Physiol.
24:69-88[CrossRef].
|
| 18.
|
Mimura, A.,
T. Kawano, and K. Yamaga.
1969.
Application of microorganisms to petrochemical industry. (I) Assimilation of nitrile compounds by microorganisms.
J. Ferment. Technol.
47:631-638.
|
| 19.
|
Nagasawa, T.,
C. D. Mathew,
J. Mauger, and H. Yamada.
1988.
Nitrile hydratase-catalyzed production of nicotinamide from 3-cyanopyridine in Rhodococcus rhodochrous J1.
Appl. Environ. Microbiol.
54:766-1769.
|
| 20.
|
Rajagopal, R., and P. Larsen.
1972.
Metabolism of indole-3-acetaldoxime in plants.
Planta
103:45-54[CrossRef].
|
| 21.
|
Sibbesen, O.,
B. Koch,
P. Rouzé,
B. L. Møller, and B. A. Halkier.
1995.
Biosynthesis of cyanogenic glucosides. Elucidation of the pathway and characterization of the cytochrome P-450 involved, p. 227-241.
In
R. M. Wallsgrove (ed.), Amino acids and their derivatives in higher plants. Cambridge University Press, Cambridge, England.
|
| 22.
|
Tani, Y.,
M. Kurihara,
H. Nishise, and K. Yamamoto.
1989.
Bioconversion of dinitrile to mononitrile, a tranexamic acid intermediate, by Corynebacterium sp.
Agric. Biol. Chem.
53:3143-3149.
|
| 23.
|
Watanabe, I.,
Y. Satoh, and K. Enomoto.
1987.
Screening, isolation and taxonomical properties of microorganisms having acrylonitrile-hydrating activity.
Agric. Biol. Chem.
51:3193-3199.
|
| 24.
|
Yamada, H.,
Y. Asano,
T. Hino, and Y. Tani.
1979.
Microbial utilization of acrylonitrile.
J. Ferment. Technol.
57:8-14.
|
| 25.
|
Yamada, H.,
Y. Asano, and Y. Tani.
1980.
Microbial utilization of glutaronitrile.
J. Ferment. Technol.
58:495-500.
|
| 26.
|
Yamada, H., and M. Kobayashi.
1996.
Nitrile hydratase and its application to industrial production of acrylamide.
Biosci. Biotechnol. Biochem.
60:1391-1400[Medline].
|
| 27.
|
Yamamoto, K.,
Y. Ueno,
K. Otsubo,
K. Kawakami, and K. Komatsu.
1990.
Production of S-(+)-ibuprofen from a nitrile compound by Acinetobacter sp. strain AK226.
Appl. Environ. Microbiol.
56:3125-3129[Abstract/Free Full Text].
|
| 28.
|
Yamamoto, K.,
K. Oishi,
I. Fujimatsu, and K. Komatsu.
1991.
Production of R-( )-mandelic acid from mandelonitrile by Alcaligenes faecalis ATCC 8750.
Appl. Environ. Microbiol.
57:3028-3032[Abstract/Free Full Text].
|
Applied and Environmental Microbiology, June 2000, p. 2290-2296, Vol. 66, No. 6
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Brandao, P. F. B., Clapp, J. P., Bull, A. T.
(2003). Diversity of Nitrile Hydratase and Amidase Enzyme Genes in Rhodococcus erythropolis Recovered from Geographically Distinct Habitats. Appl. Environ. Microbiol.
69: 5754-5766
[Abstract]
[Full Text]
-
Oinuma, K.-I., Hashimoto, Y., Konishi, K., Goda, M., Noguchi, T., Higashibata, H., Kobayashi, M.
(2003). Novel Aldoxime Dehydratase Involved in Carbon-Nitrogen Triple Bond Synthesis of Pseudomonas chlororaphis B23: SEQUENCING, GENE EXPRESSION, PURIFICATION, AND CHARACTERIZATION. J. Biol. Chem.
278: 29600-29608
[Abstract]
[Full Text]
Top
Abstract
Introduction
