Yield and rheological properties of exopolysaccharide from a local isolate: Xanthomonas axonopodis pv. vesicatoria

Background: The aim of the present study was to evaluate gum productivity of a local strain, Xanthomonas axonopodis pv. vesicatoria, isolated from pepper plant, and its rheological behavior for the first time compared to the standard strain, Xanthomonas campestris DSM 19000 (NRRL B-1459). The influence of operational conditions (agitation rate and inoculum volume) on gum production and rheological properties of gums from the Xanthomonas strains were investigated. Results: The isolated strain of Xanthomonas showed similar xanthan yield compared to the standard strain. Furthermore, this study clearly confirmed that gum yield depended on bacterial strain, agitation rate, and inoculum size. The most suitable conditions for the gum production in an orbital shaker in terms of agitation rate and inoculum size were 180 rpm and 5%, respectively, resulting in an average production of 10.96 and 11.19 g/L for X. axonopodis pv.vesicatoria and X. campestris DSM 19000, respectively. Regarding the rheological properties, Ostwald-de-Waele and power law models were used to describe flow and oscillatory behavior of the gum solutions, respectively. Consistency of the novel gum solution remarkably was much higher than the commercial xanthan gum solution. Flow and oscillatory behavior and their temperature ramps showed that weak gel-like structure could be obtained with less gum concentrations when the novel gum was used. Conclusion: Therefore, yield and technological properties of the aqueous solutions of the exopolysaccharide synthesized by X. axonopodis pv. vesicatoria were observed to be more suitable for industrial production.

Structure of xanthan gum and cell ultrastructure at different times of alkali stress

Abstract The effect of alkali stress on the yield, viscosity, gum structure, and cell ultrastructure of xanthan gum was evaluated at the end of fermentation process of xanthan production by Xanthomonas campestris pv. manihotis 280-95. Although greater xanthan production was observed after a 24 h-alkali stress process, a lower viscosity was observed when compared to the alkali stress-free gum, regardless of the alkali stress time. However, this outcome is not conclusive as further studies on gum purification are required to remove excess sodium, verify the efficiency loss and the consequent increase in the polymer viscosity. Alkali stress altered the structure of xanthan gum from a polygon-like shape to a star-like form. At the end of the fermentation, early structural changes in the bacterium were observed. After alkali stress, marked structural differences were observed in the cells. A more vacuolated cytoplasm and discontinuities in the membrane cells evidenced the cell lysis. Xanthan was observed in the form of concentric circles instead of agglomerates as observed prior to the alkali stress.

Establishment of an inducing medium for type III effector secretion in Xanthomonas campestris pv. campestris

It is well known that the type III secretion system (T3SS) and type III (T3) effectors are essential for the pathogenicity of most bacterial phytopathogens and that the expression of T3SS and T3 effectors is suppressed in rich media but induced in minimal media and plants. To facilitate in-depth studies on T3SS and T3 effectors, it is crucial to establish a medium for T3 effector expression and secretion. Xanthomonas campestris pv. campestris (Xcc) is a model bacterium for studying plant-pathogen interactions. To date no medium for Xcc T3 effector secretion has been defined. Here, we compared four minimal media (MME, MMX, XVM2, and XOM2) which are reported for T3 expression induction in Xanthomonas spp. and found that MME is most efficient for expression and secretion of Xcc T3 effectors. By optimization of carbon and nitrogen sources and pH value based on MME, we established XCM1 medium, which is about 3 times stronger than MME for Xcc T3 effectors secretion. We further optimized the concentration of phosphate, calcium, and magnesium in XCM1 and found that XCM1 with a lower concentration of magnesium (renamed as XCM2) is about 10 times as efficient as XCM1 (meanwhile, about 30 times stronger than MME). Thus, we established an inducing medium XCM2 which is preferred for T3 effector secretion in Xcc.

Influence of agitation and aeration in xanthan production by Xanthomonas campestris pv pruni strain 101 / Influencia de la agitación y la aireación en la producción de xantano por Xanthomonas campestris pv. pruni cepa 101

Production, viscosity, and chemical composition of xanthan synthesized by bacterium Xanthomonas campestris pv pruni strain 101 were evaluated in bioreactor systems. During the process, the volumetric oxygen mass transfer coefficient (kLa) and the biomass were determined and the pH was monitored. The cultures were grown in a 3 l bioreactor, with aeration and agitation varying as follows: conditions (A) 300 rpm, 3 vvm and (B) 200 rpm, 2 vvm, at 28 °C. Our results showed that gum production was dependent on kLa, with a maximum yield of 8.15 g/l at 300 rpm, 3 vvm, 54 h of fermentation, kLa 21.4/h, while biomass was not affected. All aqueous solutions of 3% (w/v) xanthans synthesized showed a pseudoplastic behavior. The highest viscosity was reached under the strongest aeration/agitation conditions. All xanthan samples contained glucose, mannose, rhamnose, and glucuronic acid as their main components. The highest agitation and aeration rates used under condition A (300 rpm and 3 vvm) favorably influenced the yield and viscosity of the xanthan produced by bacterium X. campestris pv pruni 101 at different fermentation times.

Se evaluó la producción, viscosidad y composición química del xantano sintetizado por la bacteria Xanthomonas campestris pv. pruni cepa 101 en un fermentador. Durante el proceso se controló el pH y se determinaron el coeficiente de transferencia de masa de oxígeno (kLa) y la producción de masa celular seca. Los cultivos se realizaron en un fermentador de 3 l variando la aireación y la agitación, en las siguientes condiciones: (A) 300 rpm, 3 vvm y (B) 200 rpm, 2 vvm; a 28 °C. Nuestros resultados mostraron que la producción de goma fue dependiente del kLa, con un rendimiento máximo de 8,15 g/l a 300 rpm y 3 vvm a las 54 h de fermentación, kLa de 21,4/h, mientras que la producción de biomasa no se afectó. Todas las soluciones acuosas de xantano al 3% (m/v) sintetizadas presentaron comportamiento pseudoplástico. La mayor viscosidad se alcanzó en la condición de aireación/agitación más intensa. Todas las muestras de xantano contenían glucosa, manosa, ramnosa y ácido glucurónico como constituyentes principales. La mayor tasa de agitación y aireación utilizada en la condición A (300 rpm y 3 vvm) influyó favorablemente en el rendimiento y la viscosidad del xantano producido por la bacteria X. campestris pv. pruni 101 a diferentes tiempos de fermentación.

Mutageneisis of xanthomonas campestris and selection of strains with enhanced xanthan production

Xanthan gum is a microbial polysaccharide of great commercial importance as it has unusual rheological properties in solution and consequent range of applications. In this study, a series of mutants were isolated from Xanthomonas campestris PTCC 1473 by ethyl methanesulfonate mutagenesis. The polysaccharide yield of one mutant, XC1473E2, was 30% better than that of the parent strain. It also showed higher xanthan formation and glucose consumption rates compared to the parent strain. Xanthan produced by the mutant had enhanced viscosity, higher pseudoplasticity and larger molecular weight. Since mutant XC1473E2 appeared white on agar plates, it underwent pigment extraction with methanol. Contrary to the parent strain, the mutant showed no absorption at 443nm, i.e. the wavelength related to yellow pigment. This finding suggested that yellow pigmentation and normal xanthan biosynthesis are not necessarily concurrent. In general, mutant XC1473E2 seems to be a strain with interesting characteristics for use in commercial production of xanthan

Antagonism of Yeasts to Xanthomonas Campestris PV. Campestris on Cabbage Phylloplane in field

Twenty yeast isolates, obtained from cabbage phylloplane, were evaluated for antagonistic activity against Xanthomonas campestris pv. campestris, in field. Plants of cabbage ev. Midori were pulverized simultaneously with suspensions of antagonists and pathogen. After 10 days, plants were evaluated through percentage of foliar area with lesions. Percentage of disease severity reduction (DSR per cent) was also calculated. Yeast isolates LR32, LR42 and LR19 showed, respectively, 72, 75 and 79 (per cent) of DSR. These antagonists were tested in seven different application periods in relation to pathogen inoculation (T1=4 d before; T2=simultaneously; T3=4 d after; T4=4 d before + simultaneously; T5=4 d after + simultaneously; T6=4 d before + 4 d after; T7=4 d before + simultaneously + 4 d after). The highest DSRs were showed by LR42 (71 per cent), LR42 (67 per cent), LR35 (69 per cent) and LR19 (68 per cent) in the treatments T7, T4, T5, and T6, which significantly differed from the others. The same yeast antagonists were also tested for back rot control using different cabbage cultivars (Fuyutoyo, Master-325, Matsukaze, Midori, Sekai I and Red Winner). The DSRs varied from 58 to 61 (per cent), and there was no significant difference among cultivars

Produccion de xantano por Xanthomonas campestris en un medio de cultivo no convencional / Xanthan production by Xanthomonas campestris in no-conventional culture medium

Se determinó que X. campestris vp. Ocumo, exhibió mayor capacidad para crecer y producir xantano que las otras dos variedades de X. campestris analizadas. Utiliza una amplia diversidad de fuentes de carbohidratos. Sin embargo, la cepa no produce el exopolisacárido cuando la fuente de carbohidrato proviente de materiales ricos en lignocelulosa. El Jarabe Glucosado FAVEPRO (JGF) fue la fuente de carbono con la que se logró el mayor rendimiento de xantano (23g/l) con la mayor viscosidad (7000 cps). Las condiciones óptimas de producción determinadas en fiolas de 1 L (volumen de trabajo 10 L) fueron las siguientes: 5 por ciento de azúcares totales, 0,05 por ciento de urea como fuente de nitrógeno, 0.5 por ciento de fosfato dipotásico, pH 7.5, 10 por ciento de inóculo, temperatura 30 grados Celsius, agitación entre 250-1000 rpm y aireación entre 0.3-1.0 vvm. X. campestris vp. Ocumo, fue también capaz de producir xantano (10g/l), y aumentar la viscosidad (hasta 1500 cps) de un medio basado en el extracto ácido soluble de corteza de yuca molida (EACY).

Producción de gomas xánticas / Production of xanthan gums

En el período que va desde febrero has agosto de 1990, se realizó un relevamiento de Xanthomonas campestris en la zona de Luján, provincia de Buenos Aires, de 70 muestras de vegetales infectados. Se comprobó si las 50 cepas de Xanthomonas campestris obtenidas presentaban la capacidad de producir goma xantan, así como el porcentaje de conversión de sustrato en goma y el recuento de Xanthomonas campestris viables/ml de cultivo. Los mayores números de cepas productoras del polisacárido extracelular se obtuvieron de alfalfa. Asimismo, la Xanthomonas campestris pv. alfalfae presentó elevados números de células viables y porcentajes de conversión de glucosa en goma xántica