Food Sci. Biotechnol.
19(1): 201-206 (2010)
DOI 10.1007/s10068-010-0027-9
RESEARCH ARTICLE
Xanthan Gum Production of Xanthomonas spp. Isolated from
Different Plants
Tuncay Gumus, A. Sukru Demirci, Mustafa Mirik, Muhammet Arici, and Yesim Aysan
Received: 11 September 2009 / Revised: 18 November 2009 / Accepted: 19 November 2009 / Published Online: 28 February 2010
© KoSFoST and Springer 2010
Abstract Xanthan gum were produced from the following Introduction
Xanthomonas strains; standard strain Xanthomonas
campestris NRRL B-1459 and isolated strains Xanthomonas
arbicola pv. juglandis, Xanthomonas axonopodis pv.
vesicatoria, Xanthomonas axonopodis pv. begonia,
Xanthomonas axonopodis pv. dieffenbachia. The viscosity
features of the xanthan gums obtained were determined at
25-80ºC with different pH values and were compared to
commercial xanthan gum. Our results indicate that X.
arbicola pv. juglandis showed the highest productivity
(8.22±1.52 g/L gum). This was followed by X. axonopodis
pv. begonia (7.74±1.30 g/L gum), and the control bacterial
strain X. campestris NRRL B-1459 (7.46±0.28 g/L gum).
X. axonopodis pv. vesicatoria showed the lowest
productivity (6.40±0.55 g/L gum). No xanthan gum could
be obtained from X. axonopodis pv. dieffenbachia.
Xanthan gum produced by X. axonopodis pv. vesicatoria
showed the highest viscosity value (428 mPa·sec at 1%
solution) in all Xanthomonas strains isolated from plants.
Keywords: Xanthomonas spp., xanthan gum, viscosity
feature
Tuncay Gumus ( ), A. Sukru Demirci, Muhammet Arici
Department of Food Engineering, Faculty of Agriculture, Namik Kemal
University, 59030 Tekirdag, Turkey
Tel: +90-282-293-1442; Fax: +90-282-293-1480
E-mail: tgumus@nku.edu.tr
Mustafa Mirik
Department of Plant Protection, Faculty of Agriculture, Namik Kemal
University, 59030 Tekirdag, Turkey
Yesim Aysan
Department of Plant Protection, Faculty of Agriculture, Cukurova
University, Adana, Turkey
Xanthan gum is an important biopolymer that is produced
efficiently by Gram-negative bacteria of the genus
Xanthomonas (1) and has been widely accepted commercially
(2) with an annual worldwide production of 30,000 tons,
corresponding to a market of $408 million (3-5). Xanthan
gum, a cream-colored, odorless, free-flowing powder,
hydrates rapidly in cold and hot water to give a reliable
viscosity even at low concentrations. It is highly resistant
to enzymatic degradation, extremely stable over a wide pH
range, and forms highly pseudoplastic aqueous solutions
(6). Xanthan gum, an extracellular high molecular weight
polysaccharide, has unique rheological properties which
have led to numerous applications such as stabilizing,
viscosifying, emulsifying, thickening, and suspending
agent in many fields (7). There is considerable interest in
increased industrial production of xanthan gum owing to
its extensive applications (8,9) and wide use in a broad
range of industries, such as in food, toiletries, oil recovery,
cosmetics, water-based paints, etc (10). The major
applications of xanthan gum are in the food industry as a
suspending and thickening agent. It also has wide
applications in the chemical industry (6). In the industrial
production of xanthan gum, the growth medium should be
as economical as possible (11). Xanthan is produced by
Xanthomonas campestris, a plant-associated bacterium that
is generally pathogenic for plants belonging to the family
Brassicaceae. Xanthomonas causes a variety of diseases on
the leaves, stems, or fruits; an example is ‘black rot’ of
crucifers such as cabbage, cauliflower, or broccoli (12).
Bacterial leaf spot, caused by X. campestris pv. vitians
(Brown) Dye, is an important pathogen of lettuce (Lactuca
sativa L.) worldwide. Since 1995, serious outbreaks of the
disease occurred on lettuce cultivars grown in Ohio, USA
(13).
202
It is important to screen new xanthan gum producers of
the Xanthomonas spp. with economical and functional
properties, with studies to optimize the yields and
productivity in the fermentative processes.
In this research, we investigated xanthan gum production
yield from following the Xanthomonas strains: the standard
strain X. campestris NRRL B-1459, and the isolates
Xanthomonas arbicola pv. juglandis, Xanthomonas
axonopodis pv. vesicatoria, X. axonopodis pv. begonia, and
X. axonopodis pv. dieffenbachia. The viscosity of the gums
were analyzed at 25-80ºC, at different pH values and
compared to commercial xanthan gum.
Material and Methods
Organisms Xanthomonas arbicola pv. juglandis (Ceviz
1), X. axonopodis pv. vesicatoria (XCVA3-1), X. axonopodis
pv. begonia (Xcb-9), and X. axonopodis pv. dieffenbachia
(Xad-2) were isolated from walnut (Juglans regia L),
pepper (Capsicum annuum L), begonia (Begonia X tuber
hybrid), and anthurium (Anthurium andraenum), respectively.
Identification of the strains was initially confirmed by
morphological, biochemical, and physiological tests including
potassium hydroxide solubility for Gram reaction, catalase
reaction, oxidative/fermentative metabolism, and hypersensitive
reaction on tobacco leaves. Identification of the strains was
confirmed by fatty acid methyl ester (FAME) analysis. The
4 strains of Xanthomonas spp. were isolated in Turkey (14-
17).
X. campestris NRRL-B 1459 was obtained from the
United States Department of Agriculture, Research- Education
and Economics Agricultural Research Service, Microbial
Genomics, and Bioprocessing Research Unit.
The bacteria were grown on glucose yeast extract agar
medium containing (g/L): glucose 20.0, yeast extract 10.0,
calcium carbonate 20.0, agar 14.0 at 30oC for 48 hr, stored
at 4oC and subcultured every 3 weeks.
Inoculum preparation Actively growing cells from a
newly prepared slant were inoculated into 100 mL yeast
extract malt (YM) liquid medium (Difco, Benton
Dickenson Co., Sparks, MD, USA) containing (g/L) malt
extract 3.0, peptone 5.0, yeast extract 3.0, glucose 20.0 at
pH 7.0 in a 250-mL Erlenmeyer flask. The culture was
incubated at 28-30oC for 36-48 hr in an incubator shaker.
The liquid culture was used to inoculate the final
fermentation medium. For the gum production test, 10%
(v/v) of the inoculum was added to the production medium
(18).
Production of xanthan gum Production of xanthan gum
was carried out in a fermentor with 1.5-L work volume
T. Gumus et al.
(Model Biostat B; Sigma, Braun, Germany). As showed in
the review of Rosalam and England (10), several researchers
indicated the optimal temperature of 28-30ºC, fermentation
time of 72 hr, stirring rate of 180 rpm, and initial pH of
7.2. Based on the results presented in the literatures,
fermentation operational conditions were determined as;
temperature 30ºC, pH 7.0, stirrer speed 200 rpm, and air-
flow rate 1 mL/min for 72 hr. The productivity of the 5
species of Xanthomonas by fermentation is compared with
the fixed fermentation condition.
Fermentation medium The medium consisted of (g/L)
glucose 40.0, citric acid 2.1, KH2PO4 2.866, MgCl2 0.507,
Na2SO4 0.089, H3BO3 0.006, ZnO 0.006, FeCl3 ·6H2O
0.020, and CaCO3 0.020. Glucose was used as carbon
source for the fermentation studies (19).
Xanthan concentration The final fermentation broth
was centrifuged at 10,000 × g for 30 min at 4oC to remove
the cells. The supernatant was collected and the gum was
precipitated by using isopropanol (1:2 v/v) then filtered
through a filter paper. The gum was dried at 50-60oC
overnight. The dry weight of the gum was determined
(20,21).
Preparation of gum solution Water solutions of xanthan
of 0.25, 0.50, and 1%(w/w) were prepared as described in
Doğan et al. (22).
Viscosity measurements In this study the viscosity of
the samples were measured using vibro-viscometer (SV-
10; A & D Co., Ltd., Tokyo, Japan). This type viscometer
measures viscosity by controlling the amplitude of the
sensor plates immersed in a sample and measuring the
electric current to drive the sensor plates. To perform a
measurement, firstly the thin sensor plates were immersed
into a sample. When the spring plates were vibrated with
a uniform frequency, the amplitude varied in response to
the quantity of the frictional force produced by the viscidity
between the sensor plates and the sample. The vibro-
viscometer controls the driving electric current to vibrate
the spring plates in order to make uniform amplitude. Since
the frictional force of viscidity is directly proportional to
the viscosity, the driving electric current (driving power)
for vibrating the spring plates with a constant frequency to
make uniform amplitude is also directly proportional to the
viscosity of each sample. The vibro-viscometer measures
the driving electric current to vibrate the sensor plates with
a uniform frequency and amplitude, and then the viscosity
is given by the positive correlation between the driving
electric current and the viscosity (23).
Statistical analysis The yield data of gum production of
Xanthan Gum Production by Xanthomonas spp.
Xanthomonas spp. was obtained from 5 replicates and
viscosity data of the gums was obtained from 3 replicates.
The data were analyzed by analysis of variance (ANOVA)
using the SPSS statistical package program, and differences
among the viscosity means were compared using the
Duncan’s multiple range test (24).
Results and Discussion
Production of xanthan gum The productivities, after
fermentation, of the 5 different strains of Xanthomonas
spp. isolated from different plants are given in Table 1.
Of the 5 different bacterial species used, X. arbicola pv.
juglandis showed the highest productivity (8.22±1.52 g/L
gum), X. axonopodis pv. begonia produced an average of
7.74±1.30 g/L gum while the control bacterial strain X.
campestris NRRL B-1459 produced an average of 7.46±
0.28 g/L gum. X. axonopodis pv. vesicatoria produced an
average of 6.40±0.55 g/L gum and showed the lowest
productivity. On the other hand, X. axonopodis pv.
dieffenbachia couldn’t produce any xanthan gum. The
xanthan gum production of Xanthomonas spp. followed the
sequence: X. arbicola pv. juglandis > X. axonopodis pv.
begonia >X. campestris NRRL B-1459> X. axonopodis pv.
vesicatoria > X. axonopodis pv. dieffenbachia. Rottova et
al. (25) tested 10 strains of Xanthomonas to produce
xanthan gum in standard medium and the best ones in
terms of productivity were X. campestris pv. mangi-
feraeindicae 1230 (8.93 g/L), X. campestris pv. campestris
254 (9.49 g/L), and X. campestris pv. campestris 1078
(9.67 g/L). Other reports indicate xanthan gum production
of 15.65 (26), 30 (27), 19 (28), 30 (29), 2.3-8.3 (30), and
17.7 g/L (31). The productivity obtained in this study is
comparable to those reported by Moreira et al. (30).
Some research related to xanthan gum production using
different substrates has been reported in literature. Moreno
et al. (32) studied xanthan gum production by X.
campestris NRRL B-1459 using melon as a substrate and
the maximum gum production obtained was 1.6 g/L.
Bilanovic et al. (33) investigated the use of citrus waste as
a low cost substrate for xanthan gum production and
Table 1. Productivity (g/L) of the 5 species of Xanthomonas spp. by fermentation processes
Trial X. juglandis X. begonia X. vesicatoria
1 7.8 6.4 7.0
2 6.1 7.3 5.5
3 7.9 7.4 6.6
4 10.1 7.7 6.5
5 9.2 9.9 6.4
Average 8.22±1.52A1) 7.74±1.30B 6.40±0.55D
1)Standard errors of the mean (n =5); A,BMeans within the same line with different superscript letters are significantly different (p <0.05).
203
highest gum production was 12 g/L. Lopez et al. (2) tested
4 strains of X. campestris to produce xanthan gum using
olive mill wastewaters (OMW). The most valuable strain
was X. campestris NRRL B-1459 S4LII because of its
ability to produce xanthan using 7% of OMW as the
nutrient source, producing 7 g/L of xanthan gum. Whey
was also used in earlier studies to produce xanthan gum
reaching a maximum xanthan gum production (1.2 g/100
mL of whey) with X. campestris XLM 1521. Another
study by Molina et al. (34) described the use of whey as a
carbon source and the authors reported that the maximum
gum yield was 14 g/L. In these studies, xanthan gum
production from different agricultural waste products as
substrates was compared with the polymer produced using
glucose and sucrose as substrate. According to Rosalam
and England (10), many attempts have been reported for
optimizing variables of the xanthan gum fermentation and
glucose is still the best in term of the products yield,
supply, and the product quality.
Viscosity of xanthan gum The effect of gum concentration,
pH, and temperature on the viscosity features of xanthan
gum produced using different Xanthomonas spp. and
commercial xanthan gum are given Table 2.
Viscosity measurements for the xanthan gums obtained
in this study and commercial gum were carried out in the
temperature range of 25-80oC, and at different pH values,
using 0.25, 0.50, and 1%(w/v) aqueous xanthan gum solutions
(Table 2). Apparent viscosity (AV) values exhibited
considerable differences at low concentrations for both the
commercial gum and the gums produced in the laboratory
at the same pH and temperatures. Generally, the AV of
solutions decreased with increasing temperature at all pH
values. The effect of pH and temperature on viscosity
features of water (the control liquid) are shown in Table 3
and was determined to be statistically insignificant (p>0.05).
At a concentration of 0.25%(w/v) gum solutions, the
gum produced from X. axonopodis pv. vesicatoria reached
maximum viscosity (66.0 mPa·sec) at 25oC and pH 5.0.
The lowest average viscosity value was determined as 4.50
mPa·sec with gum obtained from X. campestris at 80oC
and pH 3.5 (p <0.05).
X. diffenbachia X. campestris NRRL B-1459
- 7.4
- 7.6
- 7.0
- 7.7
- 7.6
0 7.46±0.28C
204 T. Gumus et al.

Table 2. Effect of gum concentration, pH, and temperature on viscosity features of xanthan gum produced from different
Xanthomonas spp. and commercial xanthan gum (mPa·sec)
X. gum pH Temp. X. juglandis X. begonia X. vesicatoria X. campestris Commercial
conc. (%) (oC)
25 14.9±0.2bC1) 33.5±0.3cB 58.5±0.1bA 07.5±0.1bD 15.3±0.1fC
3.5 40 10.6±0.3eC 20.0±0.2gB 48.5±0.1dA 05.7±0.3dB 10.9±0.3hC
60 08.4±0.1fC 14.5±0.2jB 38.0±0.3fgA 05.7±0.0dB 08.0±0.1jc
80 05.7±0.2hD 10.9±0.1kB 25.3±0.1iA 04.5±0.2eE 07.2±0.1kC
25 15.0±0.3bD 38.0±0.1aB 66.0±0.3aA 07.9±0.2abE 27.2±0.1aC
40 11.6±0.3dD 31.3±0.0dB 55.4±0.1cA 07.3±0.0BcE 22.3±0.1cC
0.25 5 60 07.5±0.5gD 27.6±0.2eB 47.0±0.1eA 06.5±0.2cdE 20.1±0.0dC
80 05.0±0.1hE 18.5±0.1hB 37.4±0.0gA 05.8±0.1dD 12.8±0.1gC
25 17.0±0.1aD 36.0±0.1bB 65.5±0.1aA 08.4±0.1aE 26.1±0.2bC
40 13.2±0.1cD 23.1±0.0fC 48.0±0.3dA 07.6±0.0abE 26.5±0.1abB
7 60 10.0±0.1eC 16.8±0.1iB 38.3±0.1fA 07.3±0.1bcD 17.1±0.2eB
80 07.3±0.1gC 10.5±0.2kB 27.0±0.1hA 06.5±0.2cdD 10.1±0.1lB
25 44.0±0.3cD .109±0.5cB .174±0.4bA 43.5±0.3dD 91.0±0.3cA
3.5 40 37.6±0.3dD 68.5±0.3iC .131±0.6eA 34.5±0.1hE 77.8±0.1eB
60 33.4±0.1fD 54.3±0.2jC .102±0.2hA 28.7±0.2jE 69.7±0.2fB
80 22.1±0.1iD 39.8±0.2kC 67.2±0.1kA 22.5±0.4kD 53.7±0.1jB
25 45.6±0.1dB .117±0.6bB .170±0.6cA 44.7±0.4cE 84.1±0.4cC
40 36.9±0.2dE 88.6±0.3eB .122±0.8fA 41.1±0.3eD 80.4±0.4dC
0.5 5 60 31.2±0.5gE 75.1±0.1gB 93.7±0.3iA 37.3±0.1gD 66.1±0.5hC
80 19.4±0.3jE 53.6±0.5jB 63.8±0.1lA 28.7±0.1jD 50.8±0.3kC
25 50.8±0.4aE .137±0.6aB .180±0.8aA 51.7±0.3aD 91.3±0.2aC
40 43.4±0.3cE .100±0.5dB .145±0.5dA 45.7±0.1bD 85.3±0.2bC
7 60 35.3±0.5eE 85.8±0.3fB .121±0.6gA 38.2±0.3fD 67.5±0.1gC
80 25.8±0.1hE 70.8±0.5hB 79.8±0.3jA 31.7±0.1iD 57.7±0.2iC
25 .140±0.5bD .342±0.9cC .415±0.6bB .110±0.6bE .450±0.7bA
3.5 40 85.6±0.2fE .265±0.8fC .361±0.8eB 92.7±0.4dD .421±0.4dA
60 60.8±0.1hE .194±0.5iC .294±0.7hB 77.6±0.3fD .351±0.6gA
80 41.2±0.1kE .111±0.8lC .184±0.5kB 51.3±0.4kD .276±0.7kA
25 .135±0.7cE .385±0.5bC .428±0.4aB .148±0.8aD .445±0.6cA
40 89.0±0.4eE .283±0.6eC .372±0.7dB 96.8±0.3cD .406±0.4fA
1 5 60 58.0±0.2iE .235±0.9hC .310±0.9gB 68.3±0.7hD .344±0.7iA
80 39.3±0.2lE .157±0.8kC .211±0.6jB 57.0±0.4jD .284±0.2jA
25 .149±0.6aD .409±0.5aB .374±0.6cC 87.8±0.2eE .452±0.5aA
40 91.7±0.5dD .302±0.5dC .320±0.6fB 71.7±0.3gE .415±0.6eA
7 60 69.5±0.4gD .261±0.8gB .255±0.5iC 64.1±0.7iE .350±0.3hA
80 43.0±0.5jD .192±0.7jB .133±0.4lC 38.6±0.5lE .276±0.6kA
1)Means within the same lineA,B and column a,b with different superscript letters are significantly different (p <0.05) (n =3); Data are mean±SE.

The apparent viscosity values of xanthan gum solutions
at 0.25% concentration exhibited viscosity values higher
than commercial gum, while the viscosity value of the gum
produced from X. campestris NRRL B-1459 was lower
than commercial gum. Similarly, the maximum viscosity of
aqueous solutions containing 0.5% of xanthan gum was
180 mPa·sec with gum produced from X. axonopodis pv.
vesicatoria at 25oC and pH 7.0. At this concentration, the
lowest average viscosity was determined as 19.4 mPa·sec
with gum obtained from X. arbicola pv. juglandis at 80oC
and pH 5.0 (p <0.05). For the 1% gum solutions, the
viscosity was in the range of 38.6 (X. campestris NRRL B-
1459, at 80oC and pH 7.0) to 428 mPa·sec (X. axonopodis
pv. vesicatoria, at 25oC and pH 5.0). Additionally, the
average viscosity value was determined as 450 mPa·sec for
the commercial gum at 25oC and pH 3.5 (p <0.05).
The viscosity of xanthan solution depends on both the
temperature at which the viscosity is measured and the
temperature at which the xanthan is dissolved. The
viscosity decreases with increasing temperature. This
Xanthan Gum Production by Xanthomonas spp.
Table 3. Effect of pH and temperature on the viscosity features
of water
pH Temperature ( C)
o
Viscosity (mPa·sec)
25 00.85±0.0 1)
40 0.95±0.0
3.5
60 0.73±0.0
80 0.57±0.0
25 1.34±0.0
40 0.73±0.0
5 strains showed lower viscosity. These gums produced can
60 0.73±0.0
80 0.48±0.0
25 1.03±0.0
40 1.05±0.0
7 and chemical properties of xanthan gum.
60 0.79±0.0
80 0.65±0.0
Data are mean±SE (n =3).
1)
behavior is fully reversible between 10 and 80oC (35). In
the current study, the viscosity values of the gum solutions
decreased with increasing temperature and, as expected,
the viscosity of the solutions increased with statistical
significance when the gum concentration was increased.
The viscosity values, however, were not influenced by
pH changes and did not show any linear correlation with
pH values. Rottova et al. (25) studied xanthan gum
production and rheological behavior using 10 different
strains of Xanthomonas and they determined the apparent
viscosity using 3% aqueous solutions of the gums at 25oC.
The gum produced by X. campestris pv. mangiferaindicae
1230 presented the best AV (approx. 1,800 mPa·sec).
Furthermore X. axonopodis pv. manihotis 1182 (approx.
1,000 mPa·sec) and Xanthomonas spp. 1537 (approx.
900 mPa·sec) also showed high viscosity. Other viscosity
values reported for X. campestris pv. campestris 254, X.
campestris pv. arracaciae, X. melonis, and X. campestris
pv. campestris 1167 were 104, 71, 108, and 173 cp
(mPa·sec) respectively.
Ashtapture and Shah (36) reported the AV of
approximately 200 cp for 0.5% solutions of the biopolymer
at 30oC. Rogovin et al. (37) studied xanthan gum
production with X. campestris. These authors used
fermentation media containing 3% dextrose, seeded with
5% inoculum of X. campestris NRRL B-1459 and cultured
the mix aerobically at 28oC for 96 hr. The viscosities of
aqueous solutions containing 1 and 2% of the polymer
were 3,000 and 11,000 cp, respectively. Yaseen et al. (38)
investigated rheological properties of 12 gum solutions at
concentrations of 0.05, 0.1, and 0.5%. The viscosities of
the xanthan gum solutions were an average of 16.62,
29.03, and 92.59 mPa·sec respectively.
In the current study, the amount and viscosity values of
xanthan gum produced using 4 strains Xanthomonas
205
isolated from different plants and the X campestris NRRL
B-1459 control strain were determined. A comparison of
the viscosity values obtained was made with a commercially
obtained gum. This research has shown that using different
strains Xanthomonas isolated from plants it is possible to
produce xanthan gum economically in a laboratory setting.
While xanthan gum produced from some strains
reported in the current study exhibited higher viscosity than
commercial gum, the gum obtained using some other
find applications in the food industry as emulsifiers,
stabilizers, and suspension agents. However, further studies
are needed to investigate the rate of production, rheological
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