ПЕРСПЕКТИВИ ВИКОРИСТАННЯ МЕТАБОЛІТІВ ACINETOBACTER CALCOACETICUS ІМВ В-7241 У СІЛЬСЬКОМУ ГОСПОДАРСТВІ
The possibility of simultaneous synthesis of surfactants with antimicrobial properties and phytohormones (auxins, cytokinins and abscisic acid) by Acinetobacter calcoaceticus ІMV B-7241 was revealed. It was shown that the survival of cells (105–107 cells/ml) of the Pseudomonas and Xanthomonas phytop...
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| Date: | 2015 |
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| Main Authors: | , , , |
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| Language: | English Ukrainian |
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Institute of Agrocultural Microbiology and Agro-industrial Manufacture of NAAS of Ukraine
2015
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| Online Access: | https://smic.in.ua/index.php/journal/article/view/141 |
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| Journal Title: | Agriciltural microbiology |
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Agriciltural microbiology| _version_ | 1871465585927258112 |
|---|---|
| author | Пирог, Т. П. Леонова, Н. О. Савенко, І. В. Іутинська, Г. О. |
| author_facet | Пирог, Т. П. Леонова, Н. О. Савенко, І. В. Іутинська, Г. О. |
| author_institution_txt_mv | [
{
"author": "Т. П. Пирог",
"institution": "Національний університет харчових технологій"
},
{
"author": "Н. О. Леонова",
"institution": "Інститут мікробіології і вірусології ім. Д. К. Заболотного НАН України"
},
{
"author": "І. В. Савенко",
"institution": "Національний університет харчових технологій"
},
{
"author": "Г. О. Іутинська",
"institution": "Інститут мікробіології і вірусології ім. Д. К. Заболотного НАН України"
}
] |
| author_sort | Пирог, Т. П. |
| baseUrl_str | https://smic.in.ua/index.php/journal/oai |
| collection | OJS |
| datestamp_date | 2026-07-22T10:10:42Z |
| description | The possibility of simultaneous synthesis of surfactants with antimicrobial properties and phytohormones (auxins, cytokinins and abscisic acid) by Acinetobacter calcoaceticus ІMV B-7241 was revealed. It was shown that the survival of cells (105–107 cells/ml) of the Pseudomonas and Xanthomonas phytopathogenic bacteria was within 0–33 % range after the treatment with surfactant solution (0.15 mg/ml) for 2 h. The dependence of qualitative and quantitative composition of auxins and cytokinins on the nature of carbon source in the medium cultivation (ethanol, glycerol, refined and frying sunflower oil) was shown. The maximum concentration of auxin (122.04 μ/l) and cytokinins (363.93 μ/l) was observed at cultivation of A. calcoaceticus ІMV B-7241 in medium with glycerol. The data obtained serve as the grounds for the development of technology of complex microbial preparations production with different biological properties for agricultural use. |
| doi_str_mv | 10.35868/1997-3004.21.18-24 |
| first_indexed | 2025-07-17T12:24:03Z |
| format | Article |
| fulltext |
PROSPECTS OF ACINETOBACTER CALCOACETICUS ІMV B-7241
METABOLITES USE IN AGRICULTURE
T. P. Pirog, N. O. Leonova, I. V. Savenko, G. O. Iutynska
In the last decade, the increasing resistance
of pathogen microorganisms to known biocides
is observed, that led to the search for new
alternative antimicrobial agents. From the
literature it is known that these agents are
microbial surfactants (SAS) [1-3]. Due to
ecological safety the SAS of microbial origin
can be used in medicine, agriculture sector and
food industry [1-5].
Mostly the literature reports of antifungal
activity of SAS on plant pathogens [6-8], but
except fungal diseases, agricultural crops are
increasingly influenced by bacterioses [9].
Earlier, we isolated oil-degrading bacteria
from oil-contaminated soil samples identified
as Acinetobacter calcoaceticus K-4 (IMV
B-7241) and the capacity of strain to synthesize
metabolites with surface-active and
emulsifying properties at various hydrophobic
and hydrophilic substrates was determined
[10-12].
In previous studies [13; 14] we also found
that the surfactants synthesized by A.
calcoaceticus IMV B-7241 possess
antimicrobial properties as for the number of
microorganisms (Bacillus subtilis BТ-2,
Esсherichia сoli ІЕМ-1, Candida tropicalis
PBT-5, Candida albicans D-6, Candida utilis
BVC-65). Meanwhile, it is shown that in some
cases the treatment of test culture suspension
with supernatant containing surfactants was
accompanied by the increase in the number of
living cells compared to their number before
the addition of surfactants [14]. Later it was
shown that aqueous phase, which remained
after the extraction of SAS from culture liquid
supernatant, stimulated the growth of some
bacteria cells. These unexpected results
enabled suggesting that investigated
strains-producers of SAS, in addition to
complex surfactants, synthesize other
biologically active compounds, including plant
hormones.
In connection with the above the objective
of this work is the study of antimicrobial action
of surfactants of A. calcoaceticus IMV B-7241
as for phyto-pathogenic bacteria, and the ability
of IMV B-7241 strain to synthesize of plant
hormones.
Materials and methods. The main object
of the research was A. calcoaceticus K-4 strain,
registered in the Depository of microorganisms
of Zabolotny Institute of Microbiology and
Virology of National Academy of Sciences of
Ukraine under the number IMV B-7241.
SAS of A. calcoaceticus IMV B-7241 were
found to contain glyco- (trehalose mono- and
dimycolates, trehalose mono- and diacelates)
and aminolipids [11].
The research used phytopathogenic
bacteria from Ukrainian collection of
microorganisms (UCM): Pectobacterium
carotovorum UCM В-1095, Pseudomonas
syringae UCM В-1027, P. syringae pv.
atrofaciens UCM В-1015, P. syringae pv.
coronafaciens UCM В-1154, Xanthomonas
campеstris pv. campestris UCM В-1049.
The objects of the study were also
phytopathogenic bacteria from the collection of
the department of pathogenic bacteria of
D. K. Zabolotny Institute of Microbiology and
Virology of NAAS of Ukraine: P. corrugate
9070, P. savastanoi pv. glycinea 8571, X.
translucens pv. translucens 7696, X.
vesicatoria 7790.
For A. calcoaceticus IMV B-7241
cultivation the culture medium of such
composition was used (g/l): (NH2)2CO – 0.35;
MgSO4-7H2O – 0.1; NaCl – 1.0; Na2HPO4 –
0.6; KH2PO4 – 0.14; pH 6.8-7.0. In addition
yeast autolysate was added to the medium -
0.5% (volume fraction) and micronutrients
solution - 0.1% (volume fraction) [11]. The
source of carbon – ethanol, glycerine, and
sunflower oil refined and wasted after meat
frying at the concentration of 1% (volume
fraction). The culture from the exponential
growth phase grown on the above medium
composition was used as inoculants which
contained 0.5% of a corresponding substrate
(carbon source). The amount of seed material
(104-105 cells/ml) made 10% of the culture
medium volume. The cultivation of bacteria
was carried in 750 ml flasks with 100 ml of
medium on a shaker (320 rev./min.) at 28-30°C
for 120 hours.
From the culture liquid supernatant
containing SAS (preparation 1), SAS
(preparation 2) was isolated by the extraction
with the mixture of chloroform and methanol in
the ratio of 2:1 (Folch mixture). The aqueous
phase which remained after the extraction of
SAS, was conventionally called preparation 3
by us.
The cells of producers were separated by
centrifugation (5000 g) for 45 min., the
supernatant (preparation 1) was further
processed. To do this, 50 ml of the supernatant
were placed in a cylindrical separating 200 ml
funnel, 50 ml of Folch mixture was added, the
funnel was closed with ground stopper and
shaken (extracted lipids) for 5 min. The
resulting mixture was left in the separating
funnel after the extraction for phase separation,
after what the bottom fraction was poured off
(organic extract 1) and the aqueous phase was
subject to re-extraction as described above.
After the phase separation the bottom fraction
was poured off, thus getting organic extract 2.
At the third stage 50 ml of Folch mixture were
added to the aqueous phase, the extraction was
performed to obtain the organic extract 3.
Extracts 1-3 were combined and evaporated on
a rotary evaporator IP 1M2 (Russia) at 50°C
and absolute pressure of 0.4 atm. to get constant
weight. The dry residue was dissolved in sterile
drinking water to the original volume.
Preparations were sterilized at 112°C for 30
min. The concentration of SAS in preparations
1 and 2 was set by weighing technique after the
extraction with Folch mixture.
The antimicrobial properties of
preparations were determined as follows. In the
initial suspension of one-day phyto-pathogenic
test cultures grown on agar medium (wort
agar-agar and meat-and-peptone agar in the
ratio 1:1) at 30°C, the number of living cells
was determined by Koch’s method
(colony-forming units, CFU/ml). Then 1.5 ml
of test culture suspension were placed in test
tubes, added preparations 1-3 in the amount of
1.5 ml each and kept within 1 and 2 hours at the
temperature optimal for growth of test culture,
and what the number of living cells was
determined.
The survival of pathogenic bacteria was
determined as the ratio of the number of cells in
variants treated with preparations 1-3 to the
number of cells in the initial suspension and
was expressed as a percentage.
Extracellular phytohormones - auxins,
cytokinins and abscisic acid (ABA) – were
isolated from A. calcoaceticus IMV B-7241
culture liquid supernatant by the method of
phytohormones redistribution in two
immiscible with each other phases of solvents:
ethyl acetate (for auxins and ABA), pH 3.0;
n-butanol (for cytokinins), pH 8.0 [15]. The
resulting extracts were evaporated in vacuum at
40-45°C, dry residue was dissolved in ethanol
and used for physical and chemical analysis of
phytohormones.
Prior purification and concentration of
phytohormones was performed on silica gel
plates “Silufol UV254” (Chemapol,
Czechoslovakia) in the mixture of solvents
used sequentially: chloroform, 12.5% solution
of aqueous ammonia, ethyl acetate, acetic acid
(20:1). Extracts of cytokinins, ABA and indole
compounds, purified in such a way, were
separated on plates with aluminium and silica
oxide (Merck, Germany) as described in the
work [16]. The numeric determination of
phytohormones was performed with a scanning
spectrodensitometer Sorbfil (Russia); synthetic
plant hormones of Sigma-Aldrich (Germany)
and Acros Organics (Belgium) were used as
standards. The number of extracellular
phytohormones was calculated in µg/l of
supernatant.
All experiments were performed in 3
repetitions, the number of parallel
measurements in experiments ranged from 3 to
5. The statistical analysis of the experimental
data was performed as previously described
[10-12]. The differences of averages were
considered probable at the significance level of
p <0.05.
The obtained results show the possibility
of using oil-containing industrial waste for
simultaneous synthesis both surfactants and
phytohormones. These data is the basis for the
implementation of non-waste technology,
giving an opportunity to get microbial
preparations with different biological
properties in one process. Thus, when SAS
preparations are obtained, precipitated cells can
be used for water purification from oil [12]; the
resulting culture liquid supernatant – for further
isolation of surfactants with antimicrobial
antiadhesive (including pathogenic bacteria)
properties [13; 14; 20]. Whereas, the aqueous
phase that remains after the extraction of SAS
contains plant hormones of auxin and cytokinin
origin, it can be used to stimulate the growth of
microorganisms and plants.
|
| id | oai:ojs2.smic.in.ua:article-141 |
| institution | Agriciltural microbiology |
| keywords_txt_mv | keywords |
| language | English Ukrainian |
| last_indexed | 2026-07-23T01:04:23Z |
| publishDate | 2015 |
| publisher | Institute of Agrocultural Microbiology and Agro-industrial Manufacture of NAAS of Ukraine |
| record_format | ojs |
| resource_txt_mv | smicinua/8e/5a622cd6a42a6105cacb05b0b7629c8e.pdf |
| spelling | oai:ojs2.smic.in.ua:article-1412026-07-22T10:10:42Z PROSPECTS OF ACINETOBACTER CALCOACETICUS ІMV B-7241 METABOLITES USE IN AGRICULTURE ПЕРСПЕКТИВИ ВИКОРИСТАННЯ МЕТАБОЛІТІВ ACINETOBACTER CALCOACETICUS ІМВ В-7241 У СІЛЬСЬКОМУ ГОСПОДАРСТВІ Пирог, Т. П. Леонова, Н. О. Савенко, І. В. Іутинська, Г. О. surfactants, antimicrobial action, phytopathogenic bacteria, phytohormones, biosynthesis поверхнево-активні речовини, антимікробна дія, фітопатогенні бактерії, фітогормони, біосинтез The possibility of simultaneous synthesis of surfactants with antimicrobial properties and phytohormones (auxins, cytokinins and abscisic acid) by Acinetobacter calcoaceticus ІMV B-7241 was revealed. It was shown that the survival of cells (105–107 cells/ml) of the Pseudomonas and Xanthomonas phytopathogenic bacteria was within 0–33 % range after the treatment with surfactant solution (0.15 mg/ml) for 2 h. The dependence of qualitative and quantitative composition of auxins and cytokinins on the nature of carbon source in the medium cultivation (ethanol, glycerol, refined and frying sunflower oil) was shown. The maximum concentration of auxin (122.04 μ/l) and cytokinins (363.93 μ/l) was observed at cultivation of A. calcoaceticus ІMV B-7241 in medium with glycerol. The data obtained serve as the grounds for the development of technology of complex microbial preparations production with different biological properties for agricultural use. Встановлено можливість одночасного синтезу штамом Acinetobacter calcoaceticus ІМВ В-7241 поверхнево-активних речовин (ПАР) з антимікробними властивостями і фітогормонів (ауксини, цитокініни й абсцизова кислота). Після обробки впродовж 2 год. розчином ПАР (0,15 мг/мл) виживання клітин (105–107 в мл) фітопатогенних бактерій родів Pseudomonas і Xanthomonas становило 0–33 %. Якісний і кількісний склад ауксинів і цитокінінів залежав від природи джерела вуглецю у середовищі культивування штаму ІМВ В-7241 (етанол, гліцерин, рафінована та відпрацьована соняшникова олія). Максимальна концентрація ауксинів (122,04 мкг/л) і цитокінінів (363,93 мкг/л) досягалася за умов росту A. calcoaceticus ІМВ В-7241 на гліцерині. Одержані дані є основою для розробки технології одержання комплексних мікробних препаратів з різними біологічними властивостями для використання у сільському господарстві. Institute of Agrocultural Microbiology and Agro-industrial Manufacture of NAAS of Ukraine 2015-08-06 Article Article Рецензована Стаття application/pdf application/pdf https://smic.in.ua/index.php/journal/article/view/141 10.35868/1997-3004.21.18-24 Agricultural microbiology; Vol. 21 (2015): Agriciltural microbiology; 18-24 Сільськогосподарська мікробіологія; Том 21 (2015): Сільськогосподарська мікробіологія; 18-24 1997-3004 10.35868/1997-3004.21 en uk https://smic.in.ua/index.php/journal/article/view/141/168 https://smic.in.ua/index.php/journal/article/view/141/169 Авторське право (c) 2015 T. P. Pirog, N. O. Leonova, I. V. Savenko, G. O. Iutynska https://creativecommons.org/licenses/by/4.0 |
| spellingShingle | поверхнево-активні речовини антимікробна дія фітопатогенні бактерії фітогормони біосинтез Пирог, Т. П. Леонова, Н. О. Савенко, І. В. Іутинська, Г. О. ПЕРСПЕКТИВИ ВИКОРИСТАННЯ МЕТАБОЛІТІВ ACINETOBACTER CALCOACETICUS ІМВ В-7241 У СІЛЬСЬКОМУ ГОСПОДАРСТВІ |
| title | ПЕРСПЕКТИВИ ВИКОРИСТАННЯ МЕТАБОЛІТІВ ACINETOBACTER CALCOACETICUS ІМВ В-7241 У СІЛЬСЬКОМУ ГОСПОДАРСТВІ |
| title_alt | PROSPECTS OF ACINETOBACTER CALCOACETICUS ІMV B-7241 METABOLITES USE IN AGRICULTURE |
| title_full | ПЕРСПЕКТИВИ ВИКОРИСТАННЯ МЕТАБОЛІТІВ ACINETOBACTER CALCOACETICUS ІМВ В-7241 У СІЛЬСЬКОМУ ГОСПОДАРСТВІ |
| title_fullStr | ПЕРСПЕКТИВИ ВИКОРИСТАННЯ МЕТАБОЛІТІВ ACINETOBACTER CALCOACETICUS ІМВ В-7241 У СІЛЬСЬКОМУ ГОСПОДАРСТВІ |
| title_full_unstemmed | ПЕРСПЕКТИВИ ВИКОРИСТАННЯ МЕТАБОЛІТІВ ACINETOBACTER CALCOACETICUS ІМВ В-7241 У СІЛЬСЬКОМУ ГОСПОДАРСТВІ |
| title_short | ПЕРСПЕКТИВИ ВИКОРИСТАННЯ МЕТАБОЛІТІВ ACINETOBACTER CALCOACETICUS ІМВ В-7241 У СІЛЬСЬКОМУ ГОСПОДАРСТВІ |
| title_sort | перспективи використання метаболітів acinetobacter calcoaceticus імв в-7241 у сільському господарстві |
| topic | поверхнево-активні речовини антимікробна дія фітопатогенні бактерії фітогормони біосинтез |
| topic_facet | surfactants antimicrobial action phytopathogenic bacteria phytohormones biosynthesis поверхнево-активні речовини антимікробна дія фітопатогенні бактерії фітогормони біосинтез |
| url | https://smic.in.ua/index.php/journal/article/view/141 |
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