Waste Management 25 (2005) 940–954

www.elsevier.com/locate/wasman

Microbial degradation of selected odorous substances

S. Rappert, R. Müller *

Biotechnology II, Technical University Hamburg-Harburg, Denickestrasse 15, 21071 Hamburg, Germany

Accepted 20 July 2005

Available online 2 September 2005

Abstract

A biological odor treatment system has several advantages compared to conventional physical and chemical treatment technol-
ogies: (1) it is highly efficient in the treatment of waste gases characterized by high flow rates and low concentrations of contami-
nants; (2) the biodegradable pollutants are completely destroyed; and (3) it has low cost [Devinny, J.S., Deshusses, M.A., Webster,
T.S., 1999. Biofiltration for Air Pollution Control. Lewis Publisher, New York, USA; Kennes, C., Veiga, M.C., 2001. Bioreactors
for waste gas treatment. Kluwer Academic Publishers, London]. Because microorganisms play the major role in the successful bio-
logical odor treatment system, the understanding of microbial degradation of the key odorants is very important. This article
describes the occurrence and the characteristics of selected key odorous compounds such as sulphides, amines, and pyrazine com-
pounds. The article reviews available information in the literature and our experimental results of microbial degradation of the
selected compounds. This is the first article that presents the isolation and characterization of bacterial strains that can utilize
dimethyl trisulfide (DMTS), triethylamine (TEA) or different pyrazines, as a sole carbon and energy source. The biological degra-
dation pathways of some of these compounds are postulated. Moreover, the influence of the presence of other odorous compounds
in the culture medium on the degradation of the target odorous compounds by the isolated bacteria is presented. The information
presented in the paper can be used to develop new systems for biological odor treatment.
Ó 2005 Elsevier Ltd. All rights reserved.

1. Introduction some volatile inorganic (VIC), as well as organic

The air pollutants that are detected most easily are
those related to smell or odor problems. Other contam-
inants leading to air pollution problems, such as carcin-
ogenic compounds, are more difficult to detect although
they are not less harmful. Their effect is often visible
only after several years (Kennes and Veiga, 2001). Odor
may be defined as a physiological stimulus of olfactory
cells in the presence of specific molecules. The nature
and concentration of molecules detected by olfactory
cells varies between individuals and with environmental
conditions, such as temperature, pressure and humidity.
According to this definition, the term odor includes
*
Corresponding author. Tel.: +49 40 428783118; fax: +49 40
428782127.
E-mail address: ru.mueller@tu-harburg.de (R. Müller).
(VOC), compounds (Kennes and Veiga, 2001). Odorous
waste gases are a special kind of air pollutants. Humans
can perceive even extremely small amounts of an odor-
ant. It is estimated that only 108 or 109 molecules of
odorant vapor in the nose is enough to trigger detection.
To put this in perspective, 1 lg of ethyl mercaptan in air
constitutes approximately 1016 molecules, 107 or 108
times the amount necessary for detection. Human acuity
to odor requires stringent levels of control of odor emis-
sions if such emissions are generated near populations of
human. To achieve acceptable odor intensities, in many
cases odor reduction efficiencies of 95–100% are re-
quired. This level of reduction of emissions is substan-
tially higher than that typically needed for control of
most gaseous emissions (Hunter and Oyama, 2000).
Odor emissions are important sources of air pollution
in many industrial plants, in particular in livestock
production, composting plants, food processing

0956-053X/$ - see front matter Ó 2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.wasman.2005.07.015
 S. Rappert, R. Müller / Waste Management 25 (2005) 940–954
plants, wastewater treatment facilities, pharmaceutical
processing, rubber processing, pulp and paper process-
ing, petroleum refining, paint finishing plants and chem-
ical production (Valentin and North, 1980; Glowiak
et al., 1985; Fouhy, 1992; Passant et al., 1992; Williams
and Miller, 1992; Luch, 1994; Kapahi and Gross, 1995;
Both, 2001; Rappert and Müller, 2005). Increasingly
stringent environmental legislation pertaining to odors
and other emissions is generating great interest in indus-
try to increase the effectiveness of waste air treatment
(Deshusses, 1997).
A number of odor reduction methods are available to
producers; each can be expected to perform differently
for different management systems. Odor reduction tech-
nology must be compatible with the management sys-
tem and management capabilities of the operation.
Likewise, if a particular technology does not blend into
a current scheme, failure to control odors adequately
may result (Powers, 1999). Several conventional physi-
cal and chemical technologies have been developed over
the past decades, and many books describing such tech-
niques have been published. However, more recently
new treatment alternatives have been tested, based on
the use of bioreactors. It very quickly appeared that
such systems present several advantages, among which
are their effectiveness on dilute waste streams and low
costs (Devinny et al., 1999; Kennes and Veiga, 2001).
Biological processes are recognized as the most compet-
itive systems for the deodorization of waste gases char-
acterized by high flow rates and low concentrations of
contaminants (Kosteltz et al., 1996; Deshusses, 1997;
Le Cloirec et al. (1999, 2001)). The treatment of pol-
luted air in bioreactors allows the complete destruction
of the contaminants, in contrast to conventional tech-
nologies like adsorption or absorption transferring the
contaminant from one phase (gas) to another (liquid
or solid). In bioreactors pollutants are oxidized by
microorganisms into innocuous products. In more clas-
sical technologies, such as thermal or catalytic incinera-
tion, pollutants are oxidized as well, but usually at
rather higher investment and operation costs (Kennes
and Veiga, 2001). Biological treatment is environmen-
tally safe technology, treatment is generally performed
at ambient temperatures and it does not generate nitro-
gen oxides or secondary waste streams (Deshusses,
1997). Organic pollutants are generally converted to
carbon dioxide and water under the metabolic action
of growing or resting microorganisms (Deshusses,
1997; Lewandowski and DeFilippi, 1998). Polluted air
streams containing biodegradable compounds, readily
soluble in water, can be effectively treated in a biofilter.
Unfortunately, biological treatment is not possible for
all types of air streams yet. Each air stream is different
in terms of composition of pollutants and in terms of
air stream conditions like temperature, relative humidity
and emission frequency. Therefore, each one needs to be
941
treated in a different way. Conventional biofilters have
been applied successfully to air streams containing
VOC and odor compounds, readily biodegradable and
with relatively high water solubility. However, conven-
tional biofilters present some limitations in treating pol-
luted air streams that contain: (1) compounds that are
difficult to degrade biologically (e.g., large molecule),
(2) compounds that are very poorly soluble in water,
(3) compounds released during the biodegradation pro-
cess and resulting a build-up of intermediates like cate-
chols, acetic acid, etc., and (4) compounds biodegraded
into acidic products (e.g., sulfur and nitrogen com-
pounds). Furthermore, conventional biofilters also have
some disadvantages such as: (1) the organic media need
to be replaced from time to time and (2) the systems
need a relatively large amount of space when compared
to other air treatment techniques. According to these
limitations and disadvantages of conventional
biofiltration, many new types of bioreactors for effective
waste gas treatment have been developed. Recently, the
information about various bioreactors for waste gas
treatment has been documented by Kennes and Veiga
(2001). Therefore, these data will not be described here.
Microorganisms can transform virtually any organic
compound, whether manmade or naturally occurring,
if the environmental conditions are suitable and the
compounds are not toxic to microorganisms. It is up
to engineers and scientists using biological treatment
techniques to manipulate, whenever possible, environ-
mental conditions (oxygen content, chemical composi-
tion, temperature, etc.) in order to effect complete
transformation to acceptable products in the most
cost-effective manner (Lewandowski and DeFilippi,
1998). The degree and rate of biodegradation is of crit-
ical importance for the feasibility of biological tech-
niques to clean up contaminated waste streams. In
general, biodegradation is dependent on three major fac-
tors: the presence of microorganisms that can degrade
the specific chemical structure; environmental condi-
tions that allow the microorganisms to grow and release
their degradation enzymes; and good physical contact
between the organic substrate and the organism. The
biodegradation of xenobiotics has been the subject of
numerous studies, which have resulted in thousands of
publications in scientific journals, books, and conference
proceedings (for example, Crawford and Crawford,
1996; Lewandowski and DeFilippi, 1998; Van Agteren
et al., 1998). These studies have led to a deeper under-
standing of the diversity of biodegradation processes.
As a result, it has become possible to enhance the rate
of degradation of recalcitrant pollutants during biologi-
cal treatment and to design completely new types of
treatment processes (Van Agteren et al., 1998). Much
work is being done to expand the range of pollutants
to which biodegradation can be applied, and to make
treatment techniques less expensive than current
942 S. Rappert, R. Müller / Waste Management 25 (2005) 940–954
biological processing methods and better applicable for
waste streams, which are difficult to handle.
While microbial treatment of pollution is not a new
topic and the knowledge of biological degradation of
various xenobiotics has been well documented, many
pollution problems remain to be solved. The biological
degradation of various odorous compounds has been
developed (Darlington et al., 2000; Deshusses, 1997;
Zhu, 2000; Le Cloirec et al., 2001; Williams, 2001).
However, the understanding of biological degradation
of some key odorous compounds still needs to be im-
proved. To do this, it is necessary to obtain microorgan-
isms that can degrade the odorous compounds and
determine the microbial degradation of these com-
pounds under various environmental factors found in
both natural conditions and in the polluted sites. This
information is important to aid our understanding of
biodegradation in natural ecosystems and to assist the
development of strategies for effective biological odor
treatment technologies.
This paper reviews the available information in the
literature and our own work on the microbial degrada-
tion of some key odorous compounds, such as volatile
organic sulfur compounds, amines, and pyrazines. The
lists of microorganisms that can degrade the selected
compounds were compiled from published data, and
when no microorganisms were reported, new bacterial
strains that can degrade the compounds were isolated
by the authors. Furthermore, the research study
determined: (1) the degradation ability of the isolated
microorganisms and (2) the influence of the presence
of other odorous compounds on the degradation of
the target odorous compound.
The materials and methods used for isolation and
characterization of the new bacterial strains are given
in Section 2. In Section 3, the information about biodeg-
radation of key odorous compounds both from the
literature and from our own experimental results is
described. Discussion about the microbial degradation
of the odorous compounds (mechanisms of degradation
and inhibitory effects) and the criteria for the successful
application of biological treatment are presented in
Section 4.
2. Materials and methods
2.1. Media and culture condition
All chemicals, of the highest purity available, were
obtained from Aldrich, Merck or Sigma. The mineral
medium M1 used for enrichment of the odorous
compounds-degrading bacteria and for cultivation of
bacteria contained (per liter) 2.5 g of K2HPO4, 1.0 g
KH2PO4, 2.0 g (NH4)2SO4, 0.2 g MgSO4 Æ 7H2O; the
pH was adjusted to 6.75. The substrates were provided
as the sole source of carbon and energy. The substrates
used were: (1) the sulfur-containing compounds: dim-
ethyldisulfide (DMDS), dimethyl trisulfide (DMTS),
furfurylmercaptan (FM), and pentanethiol (PT); (2)
the amines: triethylamine (TEA), and trimethylamine
(TMA); (3) the pyrazine-containing compounds: pyra-
zine (P), 2-methylpyrazine (MP), 2,3-dimethylpyrazine
(2,3-DMP), 2,5-dimethylpyrazine (DP), 2,6-dimethyl-
pyrazine (2,6-DMP), 2,3,5-trimethylpyrazine (TMP),
2,3,5,6-tetramethylpyrazine (TTMP), 2-ethylpyrazine
(EP), 2,3-diethylpyrazine (DEP), 2-ethyl-5(6)-methyl-
pyrazine (EMP), 2,3-diethyl-5-methylpyrazine (DM),
acetylpyrazine (AP) and 2-isobutyl-3-methoxypyrazine
(IP); and (4) the ester: ethylacrylate (ET). Soluble sub-
strates were sterilized by filtration. All substrates were
provided at 0.5 mM, except trimethylamine, which was
provided at 5.0 mM. The time course for the degrada-
tion of the odorous compounds in the shaking culture
was measured in duplicate inoculations. Unless stated
otherwise, 30 ml of liquid culture was incubated at 25
°C in a 100-ml serum bottle. The bottle was sealed with
a butyl rubber septum and aluminum crimp seal, to pre-
vent the loss of volatile substrates. The culture was stir-
red at 110 rpm.
2.2. Enrichment and isolation of odorous compounds-
degrading microorganisms
Soil samples, biofilter materials, and liquid medium
from biowashers, contaminated with the odorous com-
pounds of interest, were obtained from food industries
and agricultural plants in Germany. Enrichment was
performed in mineral medium M1 supplemented with
0.5 mM odorous substrate. Colonies of the odorous
compound-degrading bacteria were obtained by plating
the enrichment cultures on mineral medium M1 agar
plates and incubated in a sealed jar, which was saturated
with vapor of the odor substrate.
2.3. Analytical procedure
Bacterial growth was monitored routinely by record-
ing the optical density at 560 nm, and the total cell count
was determined by counting under the microscope using
a Thoma chamber. Substrate concentrations of the
odorous compounds were measured by gas chromatog-
raphy (model GC 6000 Vega Series 2, Carlo Erba Instru-
ments, Milan, Italy). One milliliter of the culture
medium was transferred into a 5-ml sample vial sealed
with butyl rubber septa and aluminum crimp seal, and
incubated in the water bath at 90 °C for 1 h. For amine
compounds, in order to release free amines from their
salts, 0.1 ml of 2.5 M NaOH was added to 1 ml of sam-
ple before incubation in the water bath. A sample of 500
ll of the gas phase was taken by a Headspace Sampler
(HS MOD.250, Carlo Erba Instrument) and injected
 S. Rappert, R. Müller / Waste Management 25 (2005) 940–954
into the GC equipped with a flame ionization detector
and a J&W fused silica capillary column DB-624
(J&W Scientific, Folsom, CA, USA). The temperatures
of the oven, of the injection port, and of the detector
were 185, 200, and 250 °C, respectively. Nitrogen gas
was used as carrier gas, at a flow rate of 20 ml/min.
3. Biodegradation of key odorous compounds
Microorganisms have evolved a diverse potential to
transform and even mineralize numerous organic
compounds of both natural and xenobiotic origin. This
section describes the occurrence and the importance of
selected odorous compounds that are often found by
many industries to cause odor problems, and the aspects
of their malodorous and toxic properties, which are
needed for abatement. These odorous compounds are:
volatile sulfur-containing compounds, especially
DMTS; amines such as dimethylamine (DMA), TMA,
diethylamine (DEA) and TEA; pyrazine compounds,
specifically alkylpyrazines such as DM and DP. Our
aim is to provide useful information for developing
strategies for effective biological odor control. Because
the oxygen concentration in most waste gases is several
orders of magnitude higher than the odorant concentra-
tion, oxygen limitation is not a problem for aerobic bio-
degradation of the odorous compounds. Therefore, in
this section, the information related to the biological
degradation of the selected key odorous compounds un-
der aerobic conditions is presented. Microorganisms
that degrade the selected compounds, from published
data and/or the newly isolated microorganisms, are
summarized. The biological degradation pathways of
some selected compounds are also described. Since the
waste gases always consist of a mixture of various odor-
ous compounds, the influence of the presence of other
Table 1
Odor thresholds of volatile sulfur containing compounds and microorganisms degrading these compoundsa
Compound Odor threshold (lg/m3) Odor description2
11
Dimethyl trisulfide 10 Decayed cabbage
Dimethyl disulfide 0.113 Decayed cabbage
Dimethyl sulfide 2.513 Decayed cabbage
Methanethiol 0.0243 Decayed cabbage
Cabon disulfide 2413 Decayed pumpkin
3
Hydrogen sulfide 0.029 Rotten eggs
a
Sources: 1Kanagawa and Mikami (1989); 2Maarse (1991); 3O
Neill and Phillips (1992); 4Visscher and Taylor (1993a); 5Visscher and Taylor
(1993b); 6Jordan et al. (1997); 7Reichert et al. (1998); 8Smet and Van Langenhove (1998a); 9Jonkers et al. (1999); 10Chung et al. (2001); 11Franzmann
et al. (2001); 12Hartikainen et al. (2001); 13Rosenfeld et al. (2001); 14Ruokojävi et al. (2001); 15Endoh et al. (2003); 16Rappert et al. (2004).
943
odorous compounds on the degradation of the target
odorous compounds by the selected microorganisms
was investigated.
3.1. Biodegradation of volatile sulfur-containing
compounds
A variety of volatile sulfur compounds such as di-
methyl trisulfide (DMTS, Me2S3), dimethyl disulfide
(DMDS, Me2S2), dimethyl sulfide (DMS, Me2S), meth-
anethiol (MeSH), carbon disulfide (CS2), and hydrogen
sulfide (H2S) are widely distributed in the environment.
These volatiles have been identified as predominant
odorants in the emission of a wide range of activities
in the bio-industry (e.g., wastewater treatment plants,
composting plants, rendering plants), wood-pulping
industry, food industry, and agricultural operations.
These compounds are produced by microbial degrada-
tion of sulfur-containing amino acids methionine,
cysteine and derivatives or by thermal decomposition
(Visscher and Taylor, 1993a; Reichert et al., 1998; Smet
and Van Langenhove, 1998a; Pinjing et al., 2001; Ruo-
kojävi et al., 2001; Rappert and Müller, 2005). Because
these sulphides have a low odor threshold (Table 1), the
occurrence of these substances in industrial emissions in
many cases causes serious odor problems for these
industries.
Generally, emissions of these compounds can effec-
tively be reduced by biofiltration. However, in the case
of organic sulfur compounds and/or a mixture with
hydrogen sulfide, the level of emission control at the
source (i.e., reduction, removal, or degradation effi-
ciency) is sometimes very low (Hirai et al., 1990; Van
Langenhove et al., 1991; Ruokojävi et al., 2001). Zhang
et al. (1991) reported a decrease in Me2S-removal effi-
ciency in a peat biofilter inoculated with Hyphomicro-
bium 155, from 80% to 25%, by the presence of
Microorganisms
Pseudonocardia asaccharolytica DSM 4424716
Pseudonocardia asaccharolytica DSM 442477,16, Hyphomicrobium spp.8,
Thiobacillus spp.4,5,8, Thiobacillus thioparus TK-m1
Pseudonocardia asaccharolytica DSM 442477,16, Hyphomicrobium spp.8,
Thiobacillus spp.4,5,8, Thiobacillus thioparus TK-m1 Thiocapsa roseopersicina9,
Pseudomonas putida DS115
Hyphomicrobium spp.8, Thiobacillus spp.5,8, Thiobacillus thioparus TK-m1
Paracoccus denitrificans6, Thiobacillus sp.12
Thiobacillus spp.12,14, Thiobacillus thioparus TK-m1Hyphomicrobium spp.14,
Xanthomonas spp.14, Methylophaga sulfidovorans14, Pseudomonas putida10
944 S. Rappert, R. Müller / Waste Management 25 (2005) 940–954
hydrogen sulfide and methanethiol due to a preferential
degradation of the latter. In a peat biofilter inoculated
with Thiobacillus thioparus DW44, however, the degra-
dation efficiency for Me2S was inhibited by the presence
of methanethiol, but accelerated by the presence of
hydrogen sulfide (Cho et al., 1991). Furthermore, it
was found that contrary to the biological removal of
H2S and numerous volatile organic compounds, the
removal efficiency for volatile organic sulfur compounds
in biological waste gas treatment systems is rather low
and variable (Tanji et al., 1989; Phae and Shoda, 1991;
Kasakura and Tatsukawa, 1995). Since the water solu-
bility and partition coefficients of methyl sulfides do
not limit the degradation process, poor reduction
efficiencies seem to be due to reasons associated with
the microorganisms and their metabolic processes. In
order to enhance the removal efficiencies towards sulfur
containing compounds by biological treatment tech-
niques, many new microbial strains capable of degrad-
ing such compounds have been isolated (De Zwart and
Kuenen, 1992; Brennan et al., 1996; Reichert et al.,
1998; Smet and Van Langenhove, 1998b). Microbial
strains, which have potential for the aerobic degradation
of the volatile sulfur-containing compounds, are shown
in Table 1.
3.1.1. Dimethyl trisulfide
Currently, there are no published data on the biodeg-
radation of DMTS. We found that Pseudonocardia
asaccharolytica DSM 44247 (Reichert et al., 1998) could
utilize DMTS as a sole source of carbon and energy
(Rappert et al., 2004). As shown in Fig. 1, during culti-
vation of P. asaccharolytica DSM 44247 in mineral med-
ium M1 supplemented with 0.5 mM DMTS, as a sole
carbon and energy source, all of the DMTS was utilized
within 2 d. The accumulation of DMDS and dimethyl
sulfide (DMS) was detected in the medium after 1 d
and 2 d, and they disappeared from the medium after
DMS DMDS DMTS
600
500
Concentration (µM)
400
300
200
100
0
0 2 4 6
Time (d)
Fig. 1. Aerobic degradation of dimethyl trisulfide (DMTS) by
Pseudonocardia asaccharolytica DSM 44247 in mineral medium M1
containing 0.5 mM DMTS as sole source of carbon and energy.
4 d of cultivation. Reichert et al. (1997) reported that
P. asaccharolytica DMS 44247 could completely oxidize
DMDS and DMS in liquid media with sulfate produc-
tion. DMDS was oxidized to 2 mol sulfate/mol DMDS
and the stoichiometry for DMS oxidation was 1 mol
sulfate/mol DMS. Therefore, it seems likely that the
degradation pathway of DMTS by P. asaccharolytica
DSM 44247 is as follows:
Me2 S3 ! Me2 S2 þ SO2
2

4 ! Me2 S þ 2SO4
! 2C1-compounds þ 3SO2

4
Hyphomicrobium spp. and Thiobacillus spp. converted
the methylated sulfur compounds (MeSH, Me2S and
Me2S2) to sulfate and carbon dioxide (De Zwart and
Kuenen, 1992; Brennan et al., 1996; Smet and Van Lan-
genhove (1998a,b)). Therefore, it is possible that the C1
compounds produced in the postulated degradation
pathway of DMTS by P. asaccharolytica DMS 44247
are carbon dioxide.
The effect of addition of other odorous compounds
on the degradation of DMTS by P. asaccharolytica
DMS 44247 was investigated. As shown in Table 2, it
was found that the degradation of DMDS and DMTS
by P. asaccharolytica DMS 44247 in shaking culture
was strongly inhibited in the presence of 0.5 mM of each
of the following compounds in the culture medium:
sulfur containing compounds such as FM, PT; pyrazine
compounds such as DM, DP, and IP; and ester such as
ET. The presence of amines, such as TMA and TEA, in
the culture medium exhibited no effect on DMDS degra-
dation but stimulated the degradation of DMTS by
P. asaccharolytica DMS 44247.
3.2. Biodegradation of amines
Amines, such as DMA, TMA, DEA, and TEA, are
the major pollutants in the gaseous emissions of chemi-
cal industries, food industries, and agricultural opera-
tions (Tang et al., 1996; Van Agteren et al., 1998;
DMTS-Control Chang et al., 2004; Rappert and Müller, 2005). The
characteristics of these compounds are shown in Table
3. The occurrences, toxic properties and microbial deg-
radation of each of these compounds are described
below.
3.2.1. Dimethylamine
DMA is mainly used as an accelerator in rubber vul-
canization processes, during the tanning of leather, and
for production of detergents and pesticides. It is also
formed during synthesis of pesticides (tetramethylthiu-
8 10 12 ram disulfide, TMTD) and solvents (N,N-dimethylform-
amide) (Urakami et al., 1990; Shirkot et al., 1994).
Besides being emitted to the environment due to indus-
trial emission, DMA is also formed during the degrada-
tion of trimethylamine by several microorganisms and
 S. Rappert, R. Müller / Waste Management 25 (2005) 940–954 945

Table 2
Effect of addition of other odorous compounds on the degradation of odorous compounds by the isolated bacteriaa,b,c
Straine Compound Additional compoundd
DMDS DMTS FM PT DM DP IP TEA TMA ET
P. assac. DMDS ±


(
)

± ± (
)
P. asacc. DMTS
±




+ +

DM-8 DM (
)


± (
) (
) + + (
)
DM-11 DM ±


± ±
± ± ±
DP-45 DP ±


± ± (
) ± ± ±
A. amino. TMA



(
) (
) ± ± ±

a
Source: Adapted from Rappert et al., 2004.
b
Cultivations were done in shaking culture containing 30 ml mineral medium M1 supplemented with 0.5 mM of odorous compound (5.0 mM for
TMA) and 0.5 mM of additional odorous compound. The degradation of the major compound in the culture medium was determined every day up
to 10 days.
c
Symbols: +, increase the degradation rate; ±, no effect on the degradation; (
), slightly inhibit the degradation;
, strongly inhibit the
degradation.
d
Abbreviations: DMDS, dimethyl disulfide; DMTS, dimethyl trisulfide; FM, furfurylmercaptan; PT, pentanethiol; DM, 2,3-diethyl-5-methyl-
pyrazine; DP, 2,5-dimethylpyrazine; IP, 2-isobutyl-3-methoxypyrazine; TEA, triethylamine, TMA, trimethylamine;ET, ethylacrylate.
e
Bacterial strains: P. asacc., Pseudonocardia asaccharolytica DSM44247; DM-8, Mycobacterium fortuitum DM-8; DM-11, Mycobacterium sp.
DM-11; DP-45, Rhodococcus erythropolis DP-45; A. amino., Aminobacter aminovorans.

Table 3
Odor thresholds of amines and microorganisms degrading these aminesa
Compound Odor threshold (lg/l) Odor description Microorganisms
12
Dimethylamine (DMA) 0.13 Putrid, Fishy Arthrobacter sp.3,6, Bacillus sp.1,5, Hyphomicrobium sp.2,8,15,
Methylobacterium sp.16, Psuedomonas aminovorans4,16,
Mycobacterium sp.16, Paracoccus denitrificans16,
Methylophilus methylosporus11, Micrococus sp.7, Pseudomonas sp.3,15,19,
Paracoccus sp. T23121
Trimethylamine (TMA) 0.0004412 Ammonical, Fishy Aminobacter aminovorans20,22, Paracoccus sp. T23121,
Paracoccus aminovorans16, Pseudomonas aminovorans4,17
Hyphomicrobium sp.2,15, Micrococus sp.7
Diethylamine (DEA) 3018 Ammonical, Fishy Pseudomonas citronellolis RA122, Mycobacterium diernhoferi RA222,
Hyphomicrobium sp.9, Pseudomonas sp.10,13, Candida utilis14,
Hansenula polymorpha14
Triethylamine (TEA) 0.4812 Ammonical, Fishy Pseudomonas citronellolis RA122, Mycobacterium diernhoferi RA222
a
Sources: 1Myers and Zatmann (1971); 2Attwood and Harder (1972); 3Colby and Zatman (1973); 4Boulton et al. (1974); 5Colby and Zatman
(1975); 6Loginova and Trotsenko (1975); 7Tate and Alexander (1976); 8Meiberg and Harder (1978); 9Meiberg (1979); 10Claus and Kutzner (1981);
11
Large and Haywood (1981); 12Amoore and Hautala (1983); 13Ghisalba and Kuenzi (1983); 14Zwart and Harder (1983); 15Ghisalba et al. (1985);
16
Urakami et al. (1990); 17Gamati et al. (1991); 18Lundqvist et al. (1992); 19Shirkot et al. (1994); 20Lobo et al. (1997); 21Kim et al. (2001); 22Rappert
et al. (2004).

algae (Meiberg and Harder, 1978; Lundstrom and Rac-
icot, 1983; Van Agteren et al., 1998; Kim et al., 2001).
Thomas and Alexander (1981) reported that DMA
was formed in municipal sewage that had been amended
with creatinine. Moreover, because large amounts of
DMA are used for the production and stabilization of
herbicides and pesticides, the degradation of these com-
pounds, especially after they were applied to agricultural
fields, generated DMA that was emitted to the environ-
ment (Van Agteren et al., 1998). DMA has not only a
malodorous character, but it is important because of
its role as precursor of the carcinogenic compound
N
nitrosodimethylamine (Lundstrom and Racicot,
1983). Therefore, prompt treatment is necessary.
It was found that in agricultural soil DMA was rap-
idly metabolized by microorganisms and did not accu-
mulate (Ayanaba et al., 1973; Smith and Aubin, 1992).
The metabolism of dimethylamine in the environment
under aerobic conditions starts with conversion to
methylamine and formaldehyde (Van Agteren et al.,
1998). Many microorganisms that degrade dimethyl-
amine under aerobic conditions have been isolated and
are summarized in Table 3. The hypothetical pathway
for the aerobic microbial degradation of dimethylamine
follows the same route as the degradation of trimethyl-
amine, which is summarized in Fig. 2.
3.2.2. Trimethylamine
TMA is a malodorous compound usually produced
by microbial activities from choline, bataine or trimeth-
ylamine N-oxide present in marine fish (King, 1984;
Barrett and Kwan, 1985; Lin and Hurng, 1989; Mouné
946 S. Rappert, R. Müller / Waste Management 25 (2005) 940–954

Carnitine Protein Lectine choline

CH3
H3C N CH3

Trimethylamine
O2+NADPH
a
H2O

O
H3C N CH3
CH3
Trimethylamine N-oxide

b H2C O
Formaldehyde

S S H
H3C
N CH 3 H3C N CH3
S S N
CH 3 CH 3 Dimethylamine

Tetramethylthiuram disulfide O2+NADPH

c
H2C O
Formaldehyde
H3 C NH2
Methylamine

O2+NADPH
d

H2C O + NH3
Formaldehyde Ammonia

Further mineralization

Fig. 2. The proposed formation and degradation pathways for trimethylamine by microorganisms in the environment under aerobic conditions. The
enzymes of the pathways are: a, trimethylamine monooxygenase; b, trimethylamine N-oxide demethylase; c, dimethylamine monooxygenase;
d, methylamine monooxygenase (Van Agteren et al., 1998; Kim et al., 2001).

et al., 1999; Lopez-Caballero et al., 2001). TMA is fre-
quently found in effluents of fish-meal manufacturing
processes (Sandberg and Ahring, 1992; Hwang et al.,
1994; Rappert and Müller, 2005). In addition to the mal-
odorous property, TMA is a toxic compound because of
its tissue-corrosive and penetrative properties. TMA
inhibits the biosynthesis of macromolecules, such as
DNA, RNA, proteins, and has a teratogenic effect on
mouse embryos (Guest and Varma, 1992).
Microbial degradation of trimethylamine under aero-
bic conditions has been intensively studied (Urakami
et al., 1990; Ohara et al., 1990; Lobo et al., 1997; Kim
et al., 2001). Many microorganisms that can aerobically
degrade TMA have been isolated and they are
summarized in Table 3. In a bubble column continuous
fermentation system, the rate of TMA consumption by
Aminobacter aminovorans was 1.79 mM h
1 (Lobo
et al., 1997). We found that in a shaking culture of
A. aminovorans containing mineral medium M1 supple-
mented with 5.0 mM TMA > 95% of the TMA was
utilized after 24 h, thereafter all of the TMA disap-
peared from the medium.
The influence of the presence of other odorous com-
pounds on the degradation of TMA by A. aminovorans
 S. Rappert, R. Müller / Waste Management 25 (2005) 940–954
in a shaking culture containing mineral medium M1
supplemented with 5.0 mM TMA, as the sole carbon
and energy source, was investigated. As shown in Table
2, the TMA degradation rate was slightly retarded when
0.5 mM of pyrazine compounds such as 2,3-diethyl-5-
methylpyrazine or 2,5-dimethylpyrazine were added into
the culture medium. In the presence of 0.5 mM ethylac-
rylate in the culture medium, only about 20% of TMA
was utilized by A. aminovorans after 10 d of cultivation.
The degradation of TMA was completely inhibited in
the presence of 0.5 mM volatile sulfur containing com-
pounds, such as DMDS, DMTS, FM and PT. No effect
of the addition of 0.5 mM IP or other amines such as
TEA into the culture medium on the degradation of
TMA by A. aminovorans was found (Rappert et al.,
2004).
The degradation pathway of TMA by A. aminovorans
has been proposed. The strain oxidizes TMA to DMA
by the action of the enzyme trimethylamine
dehydrogenase, then to monomethylamine by the activ-
ity of dimethylamine monooxygenase and finally to
formaldehyde and ammonia by the action of enzyme
monomethylamine dehydrogenase before carbon
incorporation in the serine pathway (Large, 1983). The
same degradation pathway has been found with a newly
isolated denitrifying bacterium, Paracoccus sp. T231
(Kim et al., 2001). The biological degradation pathway
of TMA under aerobic conditions is summarized in
Fig. 2.
3.2.3. Diethylamine
Diethylamine is used in the rubber industry, in resin,
in coloring materials, and in pharmaceutical products
(Van Agteren et al., 1998). The compound is irritating
to the skin and to mucous membranes.
Unlike DMA and TMA, little is known about the
physiology and microbial degradation of this com-
pound. In 1979, Meiberg isolated Hyphomicrobium
strains that could grow on dimethylamine and diethyl-
amine at a concentration of 1 g l
1. Pseudomonas strains
that are able to grow on diethylamine up to 5 g l
1 have
been isolated by several groups (Ghisalba and Kuenzi,
1983; Ghisalba et al., 1985; Claus and Kutzner, 1981).
Besides bacteria, some yeasts such as Candida utilis
and Hansenula polymorpha can also degrade DEA; these
strains use the compound as a nitrogen source (Zwart
and Harder, 1983). Pietsch et al. (2001) reported that un-
der conditions similar to drinking water treatment
plants, dimethylamine had higher biodegrability than
diethylamine. No information was found about the bio-
degradation of diethylamine in the environment, soil or
water samples. It is expected that the degradation path-
way of diethylamine under aerobic conditions is similar
to that of dimethylamine (Van Agteren et al., 1998).
Microorganisms that degrade DEA under aerobic con-
ditions are shown in Table 3.
947
3.2.4. Triethylamine
TEA is widely used as catalyst for polymerisation
reactions (in urethane and epoxy resin systems) and as
solvent and corrosion inhibitor. It is also used as an
intermediate in the production of various chemicals,
including pharmaceuticals (Åkesson et al., 1988). Expo-
sure to TEA may cause adverse health effects such as
asthma and visual disturbances (Belin et al., 1983; Åkes-
son et al., 1985). Moreover, as shown in Table 3, TEA
has a very low odor threshold. Therefore, an effective
treatment of this compound is necessary.
The metabolism of TEA in man has been studied
(Åkesson et al., 1988). However, little is known about
microbial degradation of TEA. Kawahara et al. (1999)
reported that TEA was not degraded in a biodegrada-
tion test performed under anaerobic conditions. In con-
trast to the results obtained under anaerobic conditions,
Tang et al. (1996) reported that 100% removal efficiency
of TEA (with the TEA loads up to 140 g (m3 h)
1) was
observed in a laboratory-scale reactor. This bioreactor
contains a mixture of sieved compost particles (equiva-
lent diameter: 1.2–2.5 mm) and chaff particles (equiva-
lent diameter: 4–5 mm), as the filter material, and
activated sludge, obtained from the wastewater treat-
ment plant of a fertilizer factory where amines are the
major pollutant treated, as the inoculum. The removal
efficiency of the biofilter decreased as the amount of
TEA introduced into the biofilter increased above the
maximum loading capacity (140 g (m3 h)
1). Palica
and Walus (1998) showed the experimental results of
the deodorization of TEA carried out in three different
natural beds. It was found that the most effective natural
bed for the deodorization was crumbled bark from
deciduous trees. The use of processed material from
mushroom production as a biofilter medium was less
effective and the use of wheat straw was the least effec-
tive. The most decisive parameter influencing the
deodorization effectiveness was the moisture content of
the air, whilst the least decisive parameters were the
feeding and contact times. It has been proven over a
wide range of odor concentrations and air flow rates
that full removal of TEA in process air is attained when
using a biofilter medium of crumbled bark from decidu-
ous trees (Palica and Walus, 1998).
At present, there are no published data on biodegra-
dation of TEA by pure microbial strains. We isolated
two bacterial strains, which are able to use TEA as a sole
carbon and energy source. The strains were identified as
Pseudomonas citronellolis RA1 and Mycobacterium dien-
hoferi RA2, respectively. In shaking culture containing
mineral medium M1 supplemented with 0.5 mM TEA
as the sole carbon and energy source, both strains
RA1 and RA2 utilized all the TEA within 4 days. Rapid
growth was found immediately after the inoculation of
either one of the strains. Growth of RA1 and
RA2 reached the stationary phase after 1 and 2 d of
948 S. Rappert, R. Müller / Waste Management 25 (2005) 940–954
cultivation, respectively. Besides growing on TEA, the
strains could also grow in the mineral medium contain-
ing DEA, as a sole carbon and energy source (Rappert
et al., 2004). The metabolic pathways of TEA by these
strains are under investigation by the authors.
3.3. Biodegradation of pyrazines
Pyrazines are heterocyclic nitrogen-containing com-
pounds found mainly in processed food, where they are
created chemically in dry heating processes. They are also
found naturally in many vegetables, insects, terrestrial
vertebrates, and marine organisms. Some microorgan-
isms are known to produce pyrazines during their primary
or secondary metabolism (Woolfson and Rothschild,
1990; Maarse, 1991; Besson et al., 1997; Adams et al.,
2002; Beck et al., 2003; Rappert and Müller, 2005). Some
pyrazines exhibit bacterialcidal (MacDonald, 1973) or
chemiprotective (Kim et al., 1997) activities. Toxicologi-
cal studies of pyrazine derivatives and scientific data rele-
vant to the safety evaluation of the use of pyrazine
derivatives as flavoring ingredients have been intensively
reviewed by Adams et al. (2002); therefore, these data will
not be described here.
More than 70 volatile pyrazines with substituents
consisting only of carbon and hydrogen atoms, desig-
nated as alkylpyrazines, have been described (Wagner
et al., 1999). Alkylpyrazines are generally found in a
wide variety of foods, beverages and food processing
plants (Maarse, 1991; Rappert and Müller, 2005). They
are considered important trace flavor components in
many cooked, deep fat fried, roasted, and toasted foods
(Maga, 1992). These molecules are also used in the food
industry as additives for flavoring (Seitz, 1994). The for-
mation of alkylpyrazines has been widely investigated
and reviewed (Vernin and Parkanyi, 1982; Heath and
Reineccius, 1986; Ohloff et al., 1985; Amrani-Hemaimi
et al., 1995; Shu, 1999). Categorically, there are three
widely accepted mechanisms for the formation of
pyrazines: the Strecker degradation, the ammonia/
acyloin reaction, and the pyrolysis of hydroxyamino
acids (Hodge, 1967; Kato et al., 1970; Wang and Odell,
1973; Shu, 1999; Rappert and Müller, 2005). It was
found that pyrazine, methylpyrazine, ethylpyrazine,
2-ethyl-6-methylpyrazine, and 2,6-diethylpyrazine were
formed from serine, whereas 2,5-dimethylpyrazine, 2,6-
dimethylpyrazine, trimethylpyrazine, 2-ethyl-3,6-dim-
ethylpyrazine and 2-ethyl-3,5-dimethylpyrazine were
formed from threonine (Shu, 1999). The odor thresholds
(i.e., the lowest concentration at which an odor can be
detected) of 80 alkylpyrazines, most of them
synthesized, were determined by gas chromatography-
olfactometry. It was found that trimethylpyrazine had
the lowest threshold (50 ng/l air) amongst mono-, di-,
tri-, and tetramethylpyrazine. Substitution of the methyl
group in position 2 of trimethylpyrazine by an ethyl
group yielded 2-ethyl-3,5-dimethylpyrazine and resulted
in a 4500-fold lower odor threshold than that of trimeth-
ylpyrazine. Moreover, further substitution of methyl
group at position 3 with an ethyl group yielded DM,
which has an odor threshold as low as that of 2-ethyl-
3,5-dimethylpyrazine (Wagner et al., 1999). From the
aroma-extract dilution analysis (AEDA), among all of
the alkylpyrazines, DM is identified as one of the only
six alkylpyrazines that play a role in the aromas of food
(Grosch, 1993, 1994). DM plays the major role in the
aroma of a medium-roasted Arabica coffee sample
(Grosch, 2001). The compounds 2,5-Dimethylpyrazine
and tetramethylpyrazine are the main pyrazines detected
in many cocoa bean- or soybean-based fermented foods,
where they are recognized as important contributors to
their flavor (Besson et al., 1997; Rappert and Müller,
2005). Because pyrazine compounds are considered as
one of the major malodorous compounds in the exhaust
gas streams of various food industries (Rappert and
Müller, 2005; Ranau and Steinhart, 2004), a prompt
treatment of these compounds is necessary.
Little is known about microbial degradation of pyra-
zine compounds. The available literature contains only a
few reports that describe the transformation of pyrazines
to other compounds by bacterial strains. Kiener (1992)
showed that Pseudomonas putida, metabolizing toluene
via benzyl alcohol, produced 5-methylpyrazinecarboxy-
lic acid quantitatively from 2,5-dimethylpyrazine. This
strain could also transform 2,3,6-trimethylpyrazine to
5,6-dimethylpyrazine-2-carboxylic acid. It was found
that the resting cells of Pseudomonas acidovorans DSM
4746, Alcaligenes facalis DSM 6269, and Alcaligenes
eutrophus DSM 6920, grown on the appropriate pyridin-
ecarboxylic acids, were capable of oxidizing pyrazine-
carboxylic acid (Kiener et al., 1994). Tinschert et al.
(2000) reported that Ralstonia/Burkholderia sp. DSM
6920 could transform pyrazine-2-carboxylic acid and
5-methylpyrazine-2-carboxylic acid to 3-hydroxypyr-
azine-2-carboxylic acid and 3-hydroxy-5-methylpyr-
azine-2-carboxylic acid, respectively. However, among
these compounds, the strain could only grow on 5-meth-
ylpyrazine-2-carboxylic acid. In order to obtain effective
biological treatment of the air that contaminated with
pyrazines, it is necessary to have microbes that can effi-
ciently degrade these compounds. Since DM and DP
were identified as the key pyrazine compounds detected
in various food processing industries, especially for cof-
fee and chocolate processing plants, we selected these
two pyrazines as the representative of alkylpyrazines
and isolated bacteria that could utilize these compounds
as the sole carbon and energy source.
3.3.1. 2,3-Diethyl-5-methylpyrazine
Two bacterial strains, DM-8 and DM-11, that utilize
DM as a sole carbon and energy source were isolated.
Strain DM-8 was identified as Mycobacterium fortuitum.
 S. Rappert, R. Müller / Waste Management 25 (2005) 940–954 949

From the identification results, strain DM-11 merits rec- Control DP-45 Control-G DP-45-G

ognition as a novel species within the genus Mycobacte- 600 1,2E+08

rium, the strain was named Mycobacterium sp. DM-11.
500 1,0E+08
Under aerobic conditions with no additional carbon

DP Concentration (µM)

G r o wt h ( c e l l s / m l )
source, both strains utilized 0.5 mM DM in the culture 400 8,0E+07

medium within 3 d of cultivation. DM-11 exhibited faster 300 6,0E+07

growth than DM-8. For both strains, the maximum cell
200 4,0E+07
concentration was obtained after 4 d. The maximum cell
concentration of DM-11 in the culture medium was about 100 2,0E+07

4 times greater than that obtained with DM-8 (Fig. 3).

0 0,0E+00
The DM-degraders were tested as to whether or not they 0 0.5 1 1.5 2 2.5 3 3.5

could utilize other pyrazine compounds that have been Time (d)

detected in waste gases generated by the food industry
(Ranau and Steinhart, 2004) as the sole carbon and energy
source, i.e., the pyrazine compounds listed in the section
entitled materials and methods. It was found that, besides
metabolizing DM, strain DM-8 could grow also on EMP
as a sole source of carbon and energy. The degradation of
2,3,5-trimethylpyrazine by the strain DM-8 was only pos-
sible in the presence of DM in the culture medium. Strain
DM-11 could utilize 2,3,5-trimethylpyrazine, EMP, and
AP as the sole carbon and energy source for growth. In
the presence of DM in the culture medium, DM-11 could
further degrade other pyrazines such MP, 2,3-DMP, DP,
2,6-DMP, EP, and AP.
The effect of the addition of other odorous
compounds on the degradation of DM by DM-8 and
DM-11 was investigated. As shown in Table 2, the deg-
radation of DM by both of the isolated strains was
strongly inhibited when 0.5 mM of sulfur compounds
(such as DMTS, FM, or PT) was added into the culture
medium. The presence of amines (such as TEA and
TMA) in the culture medium accelerated the degrada-
tion of DM by DM-8, whereas these amines showed
no influence on the degradation of DM by DM-11. ET
slightly inhibited the DM degradation by DM-8 but
Control-G Control DM-8
600
DM Concentration (µM)
Fig. 4. Growth and degradation of 2,5-dimethylpyrazine (DP) by
Rhodococcus erythropolis DP-45. Growth conditions were the same as
described in Fig. 3, only DP was used as substrate instead of DM.
showed no effect on the degradation of DM by DM-
11. The degradation pathways of DM by DM-11 are un-
der investigation by the authors and will be published
subsequently.
3.3.2. 2,5-Dimethylpyrazine
A bacterial strain DP-45, which is able to grow
aerobically in mineral medium M1 with DP as a sole
source of carbon and energy, was isolated. The strain
was identified as Rhodococcus erythropolis. Under
aerobic conditions and with no additional carbon
source, strain DP-45 utilized 0.5 mM DP within 16 h
of cultivation. High rates of growth were found immedi-
ately after the inoculation of DP-45, and growth reached
the stationary phase after 24 h of cultivation (Fig. 4).
Besides DP, strain DP-45 could grow also on MP, 2,6-
DMP, 2,3,5-trimethylpyrazine (TMP, EP and EMP.
The degradation of other pyrazine derivatives (such as
P, DEP, 2,3-DMP, and DM) was only possible in the
presence of DP in the culture medium.
The influence of the presence of other odorous
DM-11 DM-8-G DM-11-G
compounds in the culture medium on the degradation
7,0E+07 of DP by strain DP-45 was investigated. As shown in
6,0E+07 Table 2, the degradation of 2,5-dimethylpyrazine by

500
strain DP-45 in shaking culture was strongly inhibited
G rowth (ce lls /m l)

5,0E+07
400
by the presence of 0.5 mM sulfur containing com-
4,0E+07
300 pounds, such as DMTS, FM, and PT. Other pyrazine
3,0E+07
compounds such as DM or IP showed no influence (in
200
2,0E+07 the case of DM) or only a slightly inhibited (in the case
100 1,0E+07 of IP) the degradation of DP by DP-45. The presence of
0 0,0E+00 0.5 mM DMDS, amines (TEA, TMA) or ET in the cul-
0 1 2 3 4 5 6 7
ture medium had no effect on the DP degradation by
Time (d)
DP-45.
Fig. 3. Growth and degradation of 2,3-diethyl-5-methylpyrazine
(DM) by Mycobacterium fortuitum DM-8 and Mycobacterium sp.
DM-11 in mineral medium M1 at 25 °C. Cells were incubated in closed
4. Discussion
100-ml vials containing 30 ml medium supplemented with 0.5 mM DM
and were shaken at 110 rpm on a rotary shaker. DM concentration
(solid line) was measured by Headspace-GC and cell count (dashed Public concern and awareness about the quality of
line) was determined under microscope. our environment is currently a strong driving force for
950 S. Rappert, R. Müller / Waste Management 25 (2005) 940–954
improved pollution control. Microbial control of
pollution is a highly effective, economically viable
alternative method of pollution control. Bioremediation
is becoming the technology of choice for the remediation
of many contaminated environments. Many of the
processes currently used for the treatment of various
waste effluents and contaminated environments rely
enormously on microbial activity. Although microbial
control of pollution is not a new concept and a number
of biological treatment technologies have been
developed, some pollution problems, such as the odor
problem, remain to be solved.
The successful application of biological treatment is
based on specific knowledge of the chemical structure
of the target compound(s), the use of appropriate micro-
organism(s) to attack the pollutant(s), and the operation
of the process under appropriate conditions, either
in situ or ex situ. These criteria hold true for the biolog-
ical treatment of odorous compounds. It is necessary to
understand the microbial degradation of the odorous
compounds under various conditions both in the
laboratory and, when possible, at the polluted site. This
information is very important to aid our understanding
of biological degradation of odorous compounds in
natural ecosystems and to help in the development of
effective strategies for biological treatment of odorous
compounds. The fundamental steps to accomplish this
goal are: (1) to have the right microorganisms that can
degrade the compounds and (2) to investigate the ability
of the strains to degrade the compounds under different
environmental conditions. The later fundamental step is
important because the efficiency of biodegradation de-
pends on culture conditions such as the concentration
of the compound, of oxygen, and of other nutrients,
and on pH, temperature and the presence of inhibitors
(Van Lith et al., 1990; Bohn, 1996; Deshusses, 1997;
Le Cloirec, 1998; Lewandowski and DeFilippi, 1998;
Skladany et al., 1998; Devinny et al., 1999; Ramirez-Lo-
pez et al., 2000).
It was found that P. asaccharolytica DSM 44247,
which was initially isolated as a dimethyl disulfide de-
grader (Reichert et al., 1998), could utilize DMTS as a
sole carbon and energy source. This is the first report
that described the microbial degradation of DMTS
and proposed the metabolic pathway of DMTS by
the selected strain (see Section 3.1). The DMTS was
first converted into DMDS then into DMS and finally
to sulfate and carbon dioxide. It is worthwhile to
further investigate the mechanism that the strain
employed to degrade DMTS (Me2S3) into DMDS
(Me2S2). The answer will provide us with the neces-
sary knowledge to understand the biochemistry of
the reaction and, therefore, further promote our
understanding of biological degradation of other poly-
sulfur compounds. However, this mechanism remains
to be fully described.
Although the degradation of TEA in biofilters was re-
ported, no information about the degradation of this
compound by pure microorganism has been found.
Two bacterial strains, Pseudomonas citronellolis RA1
and M. dienhoferi RA2, that are able to mineralize
TEA, as the sole source of carbon and energy, were iso-
lated in our laboratory. These strains could also grow in
the mineral medium containing diethylamine as a sole
substrate. The metabolic pathway of TEA by these bac-
terial strains is under investigation.
The pyrazines-degraders have been isolated in our
laboratory. M. fortuitum DM-8 and Mycobacterium
sp. DM-11 was isolated as strains that can utilize DM,
as a sole carbon and energy source. R. erythropolis
DP-45 was isolated as a 2,5-dimethylpyrazine degrader.
Interestingly, it was found that these strains have a
broad substrate spectrum. They can utilize several of
the tested alkylpyraines, which were detected in waste
gases from food industries (Ranau and Steinhart,
2004), as sole carbon and energy source, in addition to
their own substrate. Moreover, co-metabolic degrada-
tion of some pyrazine compounds that normally could
not be utilized as a sole substrate for growth of the
isolated strains was observed. The broad substrate
spectrum and the co-metabolic degradation ability of
these isolated strains indicate high potential for applying
them to the odor treatment facilities, because the odor-
ous waste gases usually contain a mixture of pyrazines
compounds (Rappert and Müller, 2005; Ranau and
Steinhart, 2004).
Since the waste gas streams generally contain a mix-
ture of chemical compounds, the effect of the presence
of other odorants in the culture medium on the degrada-
tion of the odorant of interest by the selected bacteria
was investigated. It was found that at the concentration
used (0.5 mM); sulfur containing compounds such as
DMDS, DMTS, FM and PT strongly inhibited the deg-
radation efficiency of the target odorants by the selected
strains. On the other hand, the presence of most of the
pyrazine compounds had no influence on degradation
of amines, ester, and most of other pyrazines. However,
all the pyrazine compounds tested strongly inhibited
degradation of sulfides by P. asaccharolytica DSM
44247. The presence of amines showed no influence or
did not accelerate the degradation rate of some odorous
compounds tested. The presence of ET had no effect on
pyrazine degradation of the pyrazines degraders, but its
presence did inhibit the degradation of DMDS and
DMTS degradation by P. asaccharolytica DSM44247.
Moreover, the presence of ET significantly inhibited
the degradation of trimethylamine by A. aminovorans.
These examples show that it is not sufficient to under-
stand the biodegradation of pure compounds by pure
cultures. From our results, we would conclude, for
example, that in a mixture of pyrazines and sulfur com-
pounds no degradation should occur. The fact that
 S. Rappert, R. Müller / Waste Management 25 (2005) 940–954
chemical compounds support or inhibit the microbial
degradation of the other chemical compounds is not
new. Nevertheless, data on mechanisms of degradation
and inhibitory effects are very important for designing
high efficiency odor treatment systems.
Acknowledgements
The financial support of the Federal Ministry of Edu-
cation and Research (BMBF) for the work presented
here is acknowledged and is a part of the project ‘‘Inno-
vative methods for the collection and reduction of odor
pollution from agriculture and food industry.’’ We
thank Dr. A. Lipski for providing P. asaccharolytica
DSM 44247.
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