Xanthan gum production by wild-type isolates of Xanthomonas campestris

Five newly-isolated strains of Xanthomonas campestris when compared with the standard strain, NRRL B-1459, showed higher broth viscosity and xanthan gum production. Evaluation of polysaccharide rheology is a very important determinant for selecting new xanthan-producing isolates.     

New Bacterial Polysaccharide from Arthrobacter

A bacterial strain (NRRL B-1973) isolated from soil at Guatemala City and tentatively identified as an Arthrobacter species produced a polysaccharide with unusual properties. Conditions were studied for the production of this microbial gum in shaken flasks and 20-liter fermentors. Suitable nutrients for optimal polysaccharide production included 3% glucose, 0.3% enzyme-hydrolyzed casein, magnesium sulfate, manganese sulfate, and potassium phosphate buffer (pH 7.0). Polysaccharide yields ranged from 40 to 45%, based on initial dextrose in the medium in 3- or 4-day fermentations. The gum was readily recovered from culture fluid by alcohol precipitation in the presence of an electrolyte. The Arthrobacter gum exhibited characteristics unique for a polyelectrolyte. Viscosity of solutions was not decreased by heating in the presence of salt, and the gum withstood a temperature of 121 C for 30 min. At polysaccharide levels above 0.75%, gels were formed when solutions were autoclaved with KCl. There was no significant change in viscosity over a pH range of 5.0 to 10.0.     

Preservation of Xanthomonas campestris on agar slopes: effects on xanthan production

Xanthomonas campestris NRRL B-1459 and a variant E2, when preserved on agar slopes (transferred monthly) over 11 months did not deteriorate in their ability to produce xanthan in quantity and quality, as determined by culture in 500-ml baffled flasks. Variations between 8 and 14% (with respect to the average) in the final xanthan concentration were observed for the E2 and B-1459 strains, respectively. A wide range of final viscosities was obtained; these were consistent with the changes in gum concentration. Differences were more likely associated with differences in fermentation kinetics rather than being inherent to the strains. The rheological quality of both polysacharides was relatively constant throughout the time of culture maintenance. Preservation of these bacteria on agar slopes was an adequate method, in contrast to previous reports. In the period studied, strain E2 produced higher gum titres and slightly lower gum quality compared to strain B-1459. Correspondence to: E. Galindo     

Chemical investigations on the gum exudate from sajna (Moringa oleifera)

The purified, whole-gum exudate from the drum-stick plant (Moringa oleifera) was found to contain l-arabinose, d-galactose, d-glucuronic acid, l-rhamnose, d-mannose, and d-xylose in the molar ratios of ～ 14.5:11.3:3:2:1:1. A homogeneous, degraded-gum polysaccharide consisting of d-galactose, d-glucuronic acid, and d-mannose in the molar ratios of ～ 11.7:3.9:1, was obtained on mild hydrolysis of the whole gum with acid. Permethylation studies were conducted on the whole gum, the degraded gum, and their carboxyl-reduced products, and the results were in good agreement with those obtained from periodate oxidation followed by Smith degradation. Also, isolation and characterization of the oligosaccharides obtained from the mother liquor during preparation of the degraded gum, and by graded hydrolysis of the degraded gum, were achieved. On the basis of the results obtained from these studies, a tentative structure was assigned to the average repeating-unit of the gum.     

Kinetics of polysaccharide B-1459 fermentation

Polysaccharide gum was made by fermentation with Xanthomonas campestris NRRL B-1459 in a medium of glucose, minerals, and distillers' solubles. The effect of distillers' solubles on growth rate can be described by the familiar saturation equation. Although a quasistoichiometric relationship was observed between nitrogen utilization and growth, total nitrogen supply was not growth limiting, nor was polymer formation growth associated. Cell growth primarily took place in the early part of the fermentation; polysaccharide biosynthesis occurred throughout the fermentation. Glucose was converted to polysaccharide at a fairly constant yield, which was 70–80% of glucose consumed, under optimum conditions. The kinetic patterns observed indicate that multistage continuous fermentation will be suitable for polysaccharide production.     

pH-Dependent interaction between sodium caseinate and xanthan gum

Xanthan gum and sodium caseinate are used to improve stability and texture of food. To investigate interactions between them, the effects of pH on structure of sodium caseinate–xanthan gum complex were analyzed. HCl titration showed that the absorbance of the mixture was different from that of sodium caseinate alone throughout the acidification, and that syneresis in the mixture was delayed in acidic pH. Rennet digestion clarified that xanthan gum retarded degradation of κ-casein at pH 2.7. Atomic force microscopy revealed that xanthan gum interaction with sodium caseinate was pH-dependent. Sodium caseinate particles were individually bound with xanthan gum at pH 6.6, and a side-by-side aggregation of sodium caseinate along xanthan gum was observed at pH 4.2. The mixture formed a network composed of rod-like fibers at pH 2.7. These results indicate that hydrophobic and electrostatic interactions play a role in the complex formation at neutral and acidic pH, respectively.     

The effect of degradation on κ-carrageenan/locust bean gum/konjac glucomannan gels at acidic pH

The feasibility of textural and rheological modification of gels containing κ-carrageenan (KC) and locust bean gum (LBG) by addition of konjac glucomannan (KGM) was investigated. Special attention was paid to the effect of polysaccharide degradation during heating at acidic pH. The general effect of polysaccharide degradation was to decrease the Young's modulus, while the fracture strain in extension was scarcely affected unless the degradation was very severe.     

Purification and characterization of a pyruvated-mannose-specific xanthan lyase from heat-stable, salt-tolerant bacteria

A xanthanase complex secreted by a consortium of heat-stable, salt-tolerant bacteria includes a lyase that specifically removes terminal pyruvated beta-d-mannose residues from the side chains of xanthan gum. The enzyme was purified to homogeneity from the culture broth following ion-exchange chromatography and gel permeation chromatography. It consists of a single subunit of molecular weight 33,000. The enzyme is stable to 55 degrees C for more than 6 h in 20 mM sodium phosphate buffer (pH 5.0) containing 0.25 M NaCl. Optimal enzyme activity was observed at 0.05 M NaCl and a pH of 5. The enzyme has a pI of 3.7. It does not remove unsubstituted terminal beta-d-mannose residues from xanthan side chains nor does it hydrolyze p-nitrophenyl-beta-d-mannose. Treatment of xanthan with purified lyase results in a polysaccharide containing side chains terminating in an unsaturated 4,5-ene-glucuronic acid.     

Xanthan biopolymer production at increase concentration by pH control

Efficient production of xanthan gum by fermentation with Xanthomonas campestris NRRL B-1459 can be accomplished at concentrations of xanthan in the fermented broth > 3%. This level of more than twice that previously attained by us results from continuously controlling the fermentation pH with alkali. Only a slight decrease in fermentation rate and yield occurs. When ammonia is used for pH control, cell production more than doubles and fermentation time is shortened. However, xanthan yield is decreased by the diversion of additional sugar to growth.     

Characterization of an extracellular polysaccharide produced by a Pseudomonas strain grown on glycerol

A new extracellular charged polysaccharide composed mainly by galactose, with lower amounts of mannose, glucose and rhamnose, was produced by the cultivation of Pseudomonas oleovorans NRRL B-14682 using glycerol as the sole carbon source. Thermal and solid-state NMR analysis showed that this polymer is essentially amorphous, with a glass transition temperature of 155.7 degrees C. The exopolysaccharide aqueous solutions have viscoelastic properties similar to that of Guar gum, but with affinity to salts as a result of its polyelectrolyte character. In addition, the exopolysaccharide has demonstrated good flocculating and emulsifying properties and film-forming capacity. These properties make this polymer a good alternative to more expensive natural polysaccharides, such as Guar gum, in several applications in the food, pharmaceutical, cosmetic, textile, paper and petroleum industries.     

Genetic stability and xanthan gum production in Xanthomonas campestris pv. campestris NRRL B1459

A transposon (Tn5-SC) was constructed that can be used to quantify genetic deletions or amplifications. This transposon was used to evaluate the genomic stability of Xanthomonas campestris pv. campestris NRRL B1459 and we found that the genome of this bacterium is as stable as other Gram-negative bacteria or even more stable. Homologous recombination between plasmid sequences was determined in strain NRRL B1459 and was found to occur at a similar level to that reported for other Gram-negative bacteria. We report here that in X.c.c. NRRL B1459 there is no straightforward correlation between the occurrence of genetic rearrangements and frequency of homologous recombination. These data are discussed with respect to the reported instability of strain NRRL B1459 for xanthan gum production.     

Rheological investigation of synergistic interactions between galactomannans and non-pectic polysaccharide fraction from water soluble yellow mustard mucilage

Synergistic interactions between galactomannans (GMs) and non-pectic polysaccharides (NPP) from yellow mustard mucilage were investigated in the present study. Structural analysis revealed that NPP was mainly composed of β-1,4-d glucosidic linkage. Four types of GMs, namely fenugreek gum (FG), guar gum (GG), tara gum (TG) and locust bean gum (LBG) with mannose to galactose ratios (M/G) of 1.2, 1.7, 3.0 and 3.7, respectively, were blended with NPP at various ratios. The viscoelastic properties of the mixtures were measured in order to evaluate the effects of GM/NPP blending ratio, M/G ratio, total polysaccharide concentration and pH on the synergistic interactions. Results revealed that at a total polysaccharide concentration of 0.5% (w/w), the highest synergism occurred at the GM/NPP blending ratio of 3/7 for all four types of GMs. The interaction between TG and NPP showed the highest synergy, followed by LBG/NPP, FG/NPP and GG/NPP. At a higher total polysaccharide concentration (1.0% w/w), the mixture of TG and NPP still exhibited the highest synergy, however, the order of synergy between NPP and other GMs was changed as FG/NPP, LBG/NPP and GG/NPP. At the total polysaccharide concentration of 0.5% and GM/NPP blending ratio of 3/7, neutral pH (pH 6.5) showed the strongest synergy compared to that at pH 2.0 and pH 12.0. The mechanism of the synergistic effects could be explained by a combination of segregative association model and junction zone model.     

Physicochemical properties of exopolysaccharide produced by Lactobacillus kefiranofaciens ZW3 isolated from Tibet kefir

An exopolysaccharide (EPS) producing strain, ZW3, was isolated from Tibet kefir grain and was identified as Lactobacillus kefiranofaciens. FT-IR spectroscopy revealed the presence of carboxyl, hydroxyl, and amide groups, which correspond to a typical heteropolymeric polysaccharide. The GC analysis of ZW3 EPS revealed that it was glucogalactan in nature. Exopolymer showed similar flocculation stability like xanthan gum but better than guar gum with a melting point of 93.38 degrees C which is lower than xanthan gum (153.4 degrees C) and guar gum (490.11 degrees C). Compared with other commercially available hydrocolloids like xanthan gum, guar gum and locust gum ZW3 EPS showed much better emulsifying capability.     

Degradation of raw feather by a novel high molecular weight extracellular protease from newly isolated Bacillus cereus DCUW

Biotreatment of feather wastes and utilization of the degraded products in feed and foodstuffs has been a challenge. In the present study, we have demonstrated the degradation of feather waste by Bacillus cereus DCUW strain isolated during a functional screening based microbial diversity study on East Calcutta Wetland Area. A high molecular weight keratinolytic protease from feather degrading DCUW strain was purified and characterized. Moreover, utilization of degraded products during feather hydrolysis was developed and demonstrated. The purified keratinolytic protease was found to show pH and temperature optima of 8.5 and 50 degrees C, respectively. PMSF was found to inhibit the enzyme completely. The purified enzyme showed molecular weight of 80 kDa (from SDS-PAGE). The protease was found to have broad range substrate specificities that include keratin, casein, collagen, fibrin, BAPNA and gelatin. The protease was identified as minor extracellular protease (Vpr) by RT-PCR and northern blotting techniques. This is the first report describing the characterization of minor extracellular protease (Vpr) and its involvement in feather degradation in B. cereus group of organisms.     

An anionic galactomannan polysaccharide gum from a newly-isolated lactose-utilizing bacterium. I. Strain description and gum characterization.

As part of an effort to obtain microorganisms able to produce polysaccharide gums from whey and whey permeate, soil samples from farm fields regularly treated with whey were screened for bacteria able to produce gums from lactose. The most promising organism isolated (ATCC 55046) is a facultative anaerobe, tentatively identified as a new Erwinia species on the basis of biochemical and morphological tests. The organism produces a polysaccharide gum from lactose and other sugars (herein named lactan gum) composed of mannose, galactose, and galacturonic acid with an approximate molar ratio of 5:3:2 and containing no organic acid modifying groups. The weight average molecular weight of the gum is approximately 7 x 10(6). Aqueous solutions of lactan gum exhibit shear-thinning and elastic flow behavior with an estimated power law model flow index of 0.26 at 1% (w/w) gum. The viscosity of aqueous 1% (w/w) lactan gum solutions is stable over a pH range of 2-11, being particularly stable in alkaline environments. Aqueous 1% (w/w) gum solutions at pH 5-11 show excellent thermostability, retaining at least 80% of the original viscosity after being heated to 121 degrees C for 15 min. These flow properties indicate potential industrial applications in food and nonfood products requiring a moderate degree of thickening, wet-end additives and coating agents for paper products, ceramics, detergents, and binders for building materials.     

一株产高温蛋白酶嗜热菌A-2的筛选鉴定及酶学特性分析

本研究为从云南腾冲热泉中分离纯化得到一株产高温蛋白酶的菌株并对其进行驯化培养,用以探究该菌株的生长条件及酶学特性,通过选择培养基筛选能够分解脱脂奶粉产蛋白酶的菌株,应用常规方法液体培养菌体,探究温度、pH、碳源、氮源对菌株生长情况的影响,并采用福林酚法测蛋白酶活性。并提取蛋白酶液对酶的最适pH、温度以及热稳定性、pH稳定性进行研究。结果发现通过含脱脂奶粉的固体培养基筛选得到一株产蛋白酶菌株A-2,经过生理生化试验和16S rDNA鉴定知该菌种属于Aneurinibacillus属。酵母粉、葡萄糖、55℃、pH值7.5分别为菌株生长的最适氮源、碳源、温度和pH。此外该菌株所产的蛋白酶最适温度为60℃,在pH值7~9具有较好的酶活性。因此,该菌株为嗜热芽孢杆菌,所产的碱性蛋白酶具有较高的耐受温度和pH稳定性,为进一步开发利用提供参考的价值。     

Influence of some physicochemical factors on the viscosity of aqueous levan solutions of Zymomonas mobilis

Zymomonas mobilis strain 113 “S” produces levan – an extracellular, viscous, biologically active, non-toxic fructose polymer with a unique structure and extraordinary properties. This polysaccharide was isolated at two different degrees of purity by alcohol precipitation from aqueous solutions and was characterized with respect to some rheological properties and stability of viscous solutions. The effects of temperature, pH and salt concentration on the viscosity of 1–3% levan solutions were examined. The viscosity of levan solutions was found to be quite stable and reversible at room temperature over a wide range of pH from 4 to 11. The viscosity was slightly affected by increased salt concentration. Levan solutions were rather stable at high temperatures (up to 70°C, 1 h, pH 6), where the viscosity could be almost completerly restored (up to 80–100%). Therefore, the degradation of the polymer structure under these conditions is probably insignificant. Temperatures of 70–100°C with a pH of less than 3.5 caused irreversible degradation of the levan structure. The above-mentioned properties of levan, obtained from Zymomonas mobilis 113 “S”, demonstrated the potential for the development of various therapeutic forms of pharmacologically-active levan and their application in medicine as well as in the food and other industries.     

出芽短梗霉Aureobasidium sp. SRF的菌株特性分析及胞外多糖结构鉴定

菌株SRF是1株从意大利树莓(Rubus corchorifolius)果实表面分离、可产胞外多糖的新菌株。在鉴定其分类归属的基础上,对其产生的胞外多糖进行了结构分析和发酵条件优化,为寻找微生物多糖提供新的菌株,为开发利用资源微生物提供借鉴。通过形态学和ITS序列对比分析进行菌株鉴定;通过薄层层析和红外光谱分析,确定胞外多糖结构;通过单因素检测试验,确定影响产糖量的主要因素;响应面Plackett-Burman和Box-Behnken设计筛选发酵产胞外多糖的最优条件。结果表明,出发菌株SRF隶属于出芽短梗霉属,命名为Aureobasidium sp. SRF;SRF所产胞外多糖为普鲁兰多糖;单因素检测表明,对多糖产量影响最大的因素为碳源浓度、氮源浓度、无机离子浓度,其次是碳源、氮源、无机离子、pH值;根据响应面结果确定最优发酵条件为麦芽糖8%(质量分数)、酵母提取物3%(质量分数)、钙离子0.3 g/L、pH 6,产糖量达5.93 g/L。SRF是1株来源于树莓浆果表面,可产胞外普鲁兰多糖的出芽短梗霉新菌株,是1株产微生物多糖的候选菌株。     

Production of carbohydrates by the marine diatom Chaetoceros affinis var. willei (Gran) Hustedt. II. Preliminary investigation of the extracellular polysaccharide

The isolation of an extracellular polysaccharide from cultures of Chaetoceros affinis var. willei (Gran) Hustedt is described. The polysaccharide behaved as a homogeneous, polyanionic compound in free-boundary electrophoresis at both pH 2 and 7. It contained sulphur, presumably as sulphate half ester groups (8.7% of SO₂Na), and the following monosaccharides were tentatively identified: rhamnose, fucose, arabinose, and galactose, with the two former constituting 63% of the polysaccharide preparation. The main cellular polysaccharide was a glucan and could be extracted from the cells by dilute acid. The remaining material gave, after hydrolysis, a complex mixture of monosaccharides with rhamnose as the major component. It is concluded that the extracellular polysaccharide is probably excreted from healthy cells.     

Optimization and characterization of an extracellular polysaccharide produced byPaecilomyces lilacinus

Summary A new strain of the fungusPaecilomyces lilacinus has been isolated which produces a viscous extracellular polysaccharide in a simple medium. The polysaccharide consists of glucose and galactose moieties. The viscosity of the polysaccharide was unchanged by a range of temperature and pH.