Exploiting expolysaccharides from lactic acid bacteria

Microbial exopolysaccharides (EPSs) synthesized by lactic acid bacteria (LAB) play a major role in the manufacturing of fermented dairy products. EPS production is characterized by a large variety in terms of quantity, chemical composition, molecular size, charge, type of sidechains and rigidity of the molecules. Monosaccharide unit's composition, linkages, charge and size determine the EPS' intrinsic properties and their interactions with other milk constituents. EPSs contribute to texture, mouthfeel, taste perception and stability of the final product. Furthermore, it was reported that EPS from food grade organisms, particularly LAB, have potential as food additives and as functional food ingredients with both health and economic benefits. A better understanding of structure-function relationships of EPS in a dairy food matrix and of EPS biosynthesis remain two major challenges for further applications of EPS and the engineering of functional polysaccharides.     

Biosynthesis, characterisation, and design of bacterial exopolysaccharides from lactic acid bacteria

Lactic acid bacteria (LAB) are characterised by their conversion of a large proportion of their carbon feed, fermentable sugars, to lactic acid. However, in addition to lactic acid production, the LAB are able to divert a small proportion of fermentable sugars towards the biosynthesis of exopolysaccharides (EPSs) that are independent of the cell surface and cell wall material. These microbial EPSs when suspended or dissolved in aqueous solution provide thickening and gelling properties, and, as such, there is great interest in using EPSs from food grade microorganisms (such as the LAB that are traditionally used for food fermentations) for use as thickening agents. The current review includes a brief summary of the recent literature describing features of the biosynthetic pathways leading to EPS production. Many aspects of EPS biosynthesis in LAB are still not fully understood and a number of inferences are made regarding the similarity of the pathway to those involved in the synthesis of other cell polysaccharides, e.g., cell wall components. The main body of the review will cover practical aspects concerned with the isolation and characterisation of EPS structures. In the last couple of years, a substantial number of structures have been published and a summary of the common elements of these structures is included as is a suggestion for a system for representing structures. A brief highlight of the attempts that are being made to design ‘tailor’-made polysaccharides using genetic modification and control of metabolic flux is presented.     

Structure and hydrodynamic properties of the extracellular polysaccharide from a mutant strain (RA3W) of Erwinia chrysanthemi RA3

The structure of the extracellular polysaccharide (EPS) produced by Erwinia chrysanthemi strain RA3W, a mutant strain of E. chrysanthemi RA3, has been determined using low pressure size-exclusion and anion-exchange chromatographies, high pH anion-exchange chromatography, glycosyl linkage analysis, and 1D 1H NMR spectroscopy. The polysaccharide is structurally similar, if not identical, to the family of EPS produced by such as E. chrysanthemi strains Ech9, Ech9Sm6, and SR260. The molecular weight of EPS RA3W by ultracentrifugation (sedimentation equilibrium) and light scattering is compared with those of other E. chrysanthami EPSs, as are the viscometric properties.     

Characterization of the LP28 strain-specific exopolysaccharide biosynthetic gene cluster found in the whole circular genome of Pediococcus pentosaceus

We have previously isolated a lactic acid bacterium (LAB), Pediococcus pentosaceus LP28, from the longan fruit Euphoria longana. Since the plant-derived LAB strain produces an extracellular polysaccharide (EPS), in this study, we analyzed the chemical structure and the biosynthesizing genes for the EPS.The EPS, which was purified from the LP28 culture broth, was classified into acidic and neutral EPSs with a molecular mass of about 50 kDa and 40 kDa, respectively. The acidic EPS consisted of glucose, galactose, mannose, and N-acetylglucosamine moieties. Interestingly, since pyruvate residue was detected in the hydrolyzed acidic EPS, one of the four sugars may be modified with pyruvate. On the other hand, the neutral EPS consisted of glucose, mannose, and N-acetylglucosamine; pyruvate was scarcely detected in the polysaccharide molecule.As a first step to deduce the probiotic function of the EPS together with the biosynthesis, we determined the whole genome sequence of the LP28 strain, demonstrating that the genome is a circular DNA, which is composed of 1,774,865 bp (1683 ORFs) with a GC content of 37.1%. We also found that the LP28 strain harbors a plasmid carrying 6 ORFs composed of 5366 bp with a GC content of 36.5%. By comparing all of the genome sequences among the LP28 strain and four strains of P. pentosaceus reported previously, we found that 53 proteins in the LP28 strain display a similarity of less than 50% with those in the four P. pentosaceus strains. Significantly, 4 of the 53 proteins, which may be enzymes necessary for the EPS production on the LP28 strain, were absent in the other four P. pentosaceus strains and displayed less than 50% similarity with other LAB species. The EPS-biosynthetic gene cluster detected only in the LP28 genome consisted of 12 ORFs containing a priming enzyme, five glycosyltransferases, and a putative polysaccharide pyruvyltransferase.     

Advances in bacterial exopolysaccharides: from production to biotechnological applications

A vast number of bacterial extracellular polysaccharides (EPSs) have been reported over recent decades, and their composition, structure, biosynthesis and functional properties have been extensively studied. Despite the great diversity of molecular structures already described for bacterial EPSs, only a few have been industrially developed. The main constraints to full commercialization are their production costs, mostly related to substrate cost and downstream processing. In this article, we review EPS biosynthetic and fermentative processes, along with current downstream strategies. Limitations and constraints of bacterial EPS development are stressed and correlation of bacterial EPS properties with polymer applications is emphasized.     

Growth and exopolysaccharide (EPS) production by Oenococcus oeni I4 and structural characterization of their EPSs

AIMS: To study the influence of medium constituents on growth, and exopolysaccharide (EPS) production by a strain of Oenococcus oeni. The structure of one of the EPSs has also been characterized. METHODS AND RESULTS: EPS concentration was estimated by the phenol/sulfuric acid method. After purification and fractionation of crude EPSs, the sugar composition was determined by GLC-MS of the TMS methyl glycosides. The major polysaccharide is 2-substituted-(1-3)-beta-D-glucan. This structure was determined by methylation analysis and conventional (1)H- and (13)C-nuclear magnetic resonance spectroscopy. In addition, O. oeni synthesized two heteropolysaccharides, although a lesser proportion, constituted by galactose and glucose, and one of them also showed rhamnose. The sugar source has a clear influence on growth and EPS synthesis, and EPS production was not enhanced by adding ethanol or increasing the nitrogen source. EPS biosynthesis starts in the exponential growth phase, and continued during the stationary growth phase. CONCLUSIONS: Higher EPS yields were obtained on cultures grown on glucose + fructose. O. oeni produces a beta-glucan, as the predominant EPS, and it is also able to produce two heteropolysaccharides. Significance and Impact of the Study: This work provides a better understanding of EPS synthesis by O. oeni and shows the first EPS structure described for this species.     

Structural characterization of the exocellular polysaccharides produced by Streptococcus thermophilus SFi39 and SFi12.

We investigated the structures of the exopolysaccharides (EPSs) produced by Streptococcus thermophilus SFi39 and SFi12. Both polymers were found to have molecular masses of greater than 2 x 10(6) Da. The SFi39 EPS consisted of D-glucose and D-galactose in a molar ratio of 1:1, whereas the SFi12 EPS was composed of D-galactose, L-rhamnose, and D-glucose in a molar ratio of 3:2:1. Methylation analysis of and nuclear magnetic resonance spectra recorded from the native polysaccharide, as well as oligosaccharides released by partial acid hydrolysis, allowed the complete structural determination of the SFi39 EPS, which consists of the following tetrasaccharide repeating unit: [formula: see text] Similar spectra recorded only from the native polysaccharide were sufficient to allow the structural determination of the SFi12 EPS, which consists of the following hexasaccharide repeating unit: [formula: see text] This study shows that the texturizing properties of different S. thermophilus ropy strains are based on the production of EPSs exhibiting chemical similarities but structural differences.     

Extracellular polysaccharides of a bacterium associated with a fungal canker disease of Eucalyptus sp

Extracellular polysaccharides (EPSs) produced by an Erwinia sp associated with a fungal canker disease of Eucalyptus were fractionated into one polysaccharide that was identified with that produced by Erwinia chrysanthemi strains SR260, Ech1, and Ech9, and the other distinctively different from any other EPS produced by E. chrysanthemi strains so far studied. Their structures were determined using a combination of chemical and physical techniques including methylation analysis, low pressure gel-filtration, and anion-exchange chromatographies, high-pH anion-exchange chromatography, mass spectrometry and 1D and 2D 1H NMR spectroscopy. The new polysaccharide, identified as EPS Teranera, has the following structure: [structure: see text] The molecular weights of the polysaccharides range from 3.2-6.2 x 10(5) and their hydrodynamic properties are those of polydisperse, polyanionic biopolymers with pseudoplastic, non-thixotropic flow characteristics in aqueous solutions.     

Partial characterization of extracellular polysaccharides from cyanobacteria

Four cyanobacterial strains, Cyanothece sp., Oscillatoria sp., Nostoc sp. and Nostoc carneum were studied for physico-chemical characterization of extracellular polysaccharide (EPS) secreted during the controlled growth condition. Hydrolyzed EPSs showed the compositional involvement of four sugar moieties viz. mannose, glucose, xylose and ribose in varying combinations. Infrared spectra of EPSs showed a specific absorbance of O-H stretching at 3448-3400 cm(-1), asymmetrical-symmetrical C-H stretching at 2924 and 2854 cm(-1) and a bending vibration of C-H at 1400-1380 cm(-1). Absorbance at 1259 and 1140 cm(-1) with Cyanothece sp. EPS, indicated the presence of sulfur containing functional group. Thermal gravimetric analysis and differential scanning calorimetric analysis confirmed the polysaccharides thermal stability as high as around 250 degrees C. In the presence of 0.1 M NaCl aqueous solution, the intrinsic viscosity of polysaccharides from Oscillatoria sp. and Nostoc sp. decreased 1.6 fold, whereas, 3-5 fold reduction in intrinsic viscosity was observed with commercially available guar and xanthan gum.     

Optimization of exopolysaccharide overproduction by Lactobacillus confusus in solid state fermentation under high salinity stress

It is believed that high concentrations of sodium chloride (NaCl) suppress the biosynthesis of exopolysaccharide (EPS) in lactic acid bacteria (LAB). Nevertheless, overproduction of EPSs due to high salinity stress in solid state fermentation performed on an agar surface was demonstrated in this study using a response surface methodology via a central composite design (CCD). Under optimized conditions with NaCl 4.97% and sucrose 136.5 g/L at 40.79 h of incubation, the EPS yield was 259% (86.36 g/L of EPS), higher than the maximum yield produced with the modified MRS medium containing only 120 g/L of sucrose without NaCl (33.4 g/L of EPS). Biosynthesis of EPS by Lactobacillus confusus TISTR 1498 was independent of biomass production. Our results indicated that high salinity stress can enhance EPS production in solid state fermentation.     

End plate spikes in the human electromyogram. Revision of the fusimotor theory.

'End plate spike' (EPS) is a spontaneous action potential of a normal striated muscle. EPSs are found in local 'active spots' of the muscle. The prevailing hypothesis about the origin of EPSs states that when a needle electrode affects a motor nerve branch near the neuromuscular junction at the end plate zone, an increased leakage of acetylcholine to the synaptic cleft ensues. This elicits postsynaptic action potentials of the muscle fibre which can be recorded as EPSs with the same needle electrode. Thus EPSs are thought to be caused by needle injury or irritation of the motor axon. We suggest that EPSs are action potentials of intrafusal muscle fibres and that 'active spots' are in fact muscle spindles. Waveform analysis reveals three types of EPSs: small EPSs, not propagated outside the active spot either: i) with negative onset; or ii) with short positive initial deflection; and iii) large EPSs resembling propagated motor unit potentials (MUPs) but with a typical EPS firing pattern, distinctly different from that of the MUPs. Study of EPS activation in different manoeuvres associates small EPSs with intrafusal gamma motor units and large MUP-like EPSs with beta motor units.     

Role of Streptococcus thermophilus MR-1C Capsular Exopolysaccharide in Cheese Moisture Retention

Recent work by our group has shown that an exopolysaccharide (EPS)-producing starter pair, Streptococcus thermophilus MR-1C and Lactobacillus delbrueckii subsp. bulgaricus MR-1R, can significantly increase moisture retention in low-fat mozzarella (D. B. Perry, D. J. McMahon, and C. J. Oberg, J. Dairy Sci. 80:799–805, 1997). The objectives of this study were to determine whether MR-1C, MR-1R, or both of these strains are required for enhanced moisture retention and to establish the role of EPS in this phenomenon. Analysis of low-fat mozzarella made with different combinations of MR-1C, MR-1R, and the non-EPS-producing starter culture strains S. thermophilus TA061 and Lactobacillus helveticus LH100 showed that S. thermophilus MR-1C was responsible for the increased cheese moisture level. To investigate the role of the S. thermophilus MR-1C EPS in cheese moisture retention, the epsE gene in this bacterium was inactivated by gene replacement. Low-fat mozzarella made with L. helveticus LH100 plus the non-EPS-producing mutant S. thermophilus DM10 had a significantly lower moisture content than did cheese made with strains LH100 and MR-1C, which confirmed that the MR-1C capsular EPS was responsible for the water-binding properties of this bacterium in cheese. Chemical analysis of the S. thermophilus MR-1C EPS indicated that the polymer has a novel basic repeating unit composed of d-galactose, l-rhamnose, and l-fucose in a ratio of 5:2:1.Lactic acid bacteria (LAB) are a diverse group of industrially important, gram-positive, non-spore-forming microbes that produce lactic acid as a major product of carbohydrate fermentation. Many strains of LAB produce extracellular polysaccharides which may be tightly associated with the bacterial cell wall as capsules or liberated into the growth medium as a loose slime (5). The term exopolysaccharide (EPS) has been used to refer to either type of external polysaccharide. EPSs may be homopolysaccharides, composed of a single type of sugar monomer, or heteropolysaccharides, containing several types of sugar monomers (25). Extracellular homopolysaccharides are made by such LAB as Leuconostoc mesenteroides and Streptococcus mutans, while extracellular heteropolysaccharides are produced by several other species of LAB, including Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus (6).The ability to produce EPS is unstable in LAB and may be lost following numerous transfers, prolonged periods of storage, or incubation at temperatures above that optimal for growth (6, 24). This instability of EPS production in mesophilic LAB has been attributed to the fact that the genes involved in polymer production are plasmid encoded. In contrast, genes for EPS production in thermophilic LAB, such as S. thermophilus and L. delbrueckii subsp. bulgaricus, are believed to be chromosomally encoded (6). Consequently, the unstable nature of the EPS phenotype in thermophilic strains is not understood, but it may be related to mobile genetic elements or genomic instability (24).Because of the ability of EPSs to act as viscosifying, stabilizing, or water-binding agents in various foods, these polymers can act as effective natural alternatives to commercial stabilizers (6). For example, EPS-producing (EPS⁺) LAB are commonly used as starter cultures for yogurt manufacture because EPS improves the viscosity and texture of yogurt and decreases its susceptibility to syneresis (loss of whey from the curd) (14, 28).Analysis of cheese microstructure has shown that in full-fat or part-skim mozzarella, the fat and a large portion of the water are located within channels that are formed by fat globules when the cheese curd is heated and stretched (18, 20). In low-fat mozzarella, however, there are very few fat globules to break up the protein matrix, resulting in less space for water retention (20). As a consequence, the cheese has a tough and rubbery texture and requires more heat for melting (19). Merrill et al. showed that procedures which increased moisture levels in reduced- and low-fat mozzarella improved the body, texture, and functional properties of the cheese (19). In addition to enhanced functionality, the ability to increase cheese moisture level (even by as little as 1%) gives processors an important economic advantage in the highly competitive mozzarella industry (27).Since EPS has the capacity to bind significant amounts of water, it was the hypothesis of our group that EPS⁺ LAB may be useful for the production of reduced- and low-fat mozzarella. Work by Perry et al. (21) recently showed that an EPS⁺ starter pair, S. thermophilus MR-1C and L. delbrueckii subsp. bulgaricus MR-1R, could be used to significantly increase moisture levels in low-fat mozzarella. The objectives of this study were to determine whether MR-1C, MR-1R, or both of these strains are required for enhanced moisture retention and to establish the role of EPS in this phenomenon. The results showed that S. thermophilus MR-1C was responsible for the increased cheese moisture level and demonstrated that this effect required the bacterium’s capsular EPS.(Part of this research was presented at the 92nd Annual Meeting of the American Dairy Science Association, Guelph, Ontario, Canada, 22 to 25 June 1997.)     

Optimization of Exopolysaccharide Overproduction by Lactobacillus confusus in Solid State Fermentation under High Salinity Stress

It is believed that high concentrations of sodium chloride (NaCl) suppress the biosynthesis of exopolysaccharide (EPS) in lactic acid bacteria (LAB). Nevertheless, overproduction of EPSs due to high salinity stress in solid state fermentation performed on an agar surface was demonstrated in this study using a response surface methodology via a central composite design (CCD). Under optimized conditions with NaCl 4.97% and sucrose 136.5 g/L at 40.79 h of incubation, the EPS yield was 259% (86.36 g/L of EPS), higher than the maximum yield produced with the modified MRS medium containing only 120 g/L of sucrose without NaCl (33.4 g/L of EPS). Biosynthesis of EPS by Lactobacillus confusus TISTR 1498 was independent of biomass production. Our results indicated that high salinity stress can enhance EPS production in solid state fermentation.     

Complete Genome Sequence of Streptococcus thermophilus Strain MN-ZLW-002

Streptococcus thermophilus MN-ZLW-002 was originally isolated from traditionally fermented Chinese dairy products. One of the strain-dependent characteristics of this bacterium is its ability to produce exopolysaccharides (EPSs). This study determined and analyzed the genome sequence of MN-ZLW-002. Its complete genome comprised 2,046 genes and 1,848,520 nucleotides with an average GC content of 39%. The EPS cluster of MN-ZLW-002 includes 25 open reading frames (ORFs), and some results indicate a horizontal gene transfer between MN-ZLW-002 and other lactic acid bacteria (LAB).     

A positive correlation between bacterial autoaggregation and biofilm formation in native Sinorhizobium meliloti isolates from Argentina

Sinorhizobium meliloti is a symbiotic nitrogen-fixing bacterium that elicits nodule formation on roots of alfalfa plants. S. meliloti produces two exopolysaccharides (EPSs), termed EPS I and EPS II, that are both able to promote symbiosis. EPS I and EPS II are secreted in two major fractions that reflect differing degrees of subunit polymerization, designated high- and low-molecular-weight fractions. We reported previously that EPSs are crucial for autoaggregation and biofilm formation in S. meliloti reference strains and isogenic mutants. However, the previous observations were obtained by use of "domesticated" laboratory strains, with mutations resulting from successive passages under unnatural conditions, as has been documented for reference strain Rm1021. In the present study, we analyzed the autoaggregation and biofilm formation abilities of native S. meliloti strains isolated from root nodules of alfalfa plants grown in four regions of Argentina. 16S rRNA gene analysis of all the native isolates revealed a high degree of identity with reference S. meliloti strains. PCR analysis of the expR gene of all the isolates showed that, as in the case of reference strain Rm8530, this gene is not interrupted by an insertion sequence (IS) element. A positive correlation was found between autoaggregation and biofilm formation abilities in these rhizobia, indicating that both processes depend on the same physical adhesive forces. Extracellular complementation experiments using mutants of the native strains showed that autoaggregation was dependent on EPS II production. Our results indicate that a functional EPS II synthetic pathway and its proper regulation are essential for cell-cell interactions and surface attachment of S. meliloti.     

Exopolysaccharide Synthesized by Lactobacillus reuteri Decreases the Ability of Enterotoxigenic Escherichia coli To Bind to Porcine Erythrocytes

This study investigated the therapeutic potential of bacterial polysaccharides by employing a model system based on enteroxigenic Escherichia coli (ETEC)-induced hemagglutination of erythrocytes. Exopolysaccharides produced by strains of Lactobacillus reuteri inhibited ETEC-induced hemagglutination of porcine erythrocytes. No effect was observed for dextran produced from Weissella cibaria and commercially available oligo- and polysaccharides.Lactic acid bacteria (LAB) are important in food production due to their positive contribution to flavor and preservation of the final product. Some of these food-grade microorganisms are also valuable for their ability to synthesize exopolysaccharides (EPSs). EPSs are high-molecular-weight sugar polymers, which remain attached to the microorganism as capsular EPS or become excreted into the environment in the form of slime or ropy EPS (26, 30). LAB utilize two distinct biosynthetic pathways to produce EPSs. Heteropolysaccharides (HePSs) comprised of two to eight repeating units of monosaccharides are assembled by cell wall-bound glycosyltransferases in low quantities from intracellular sugar nucleotide precursors (4). Extracellular glycansucrases (glucan- or fructansucrases) synthesize homopolysaccharides (HoPSs) consisting of either glucose or fructose from sucrose, and their yield can be as high as 40 g liter⁻¹ (13, 19). EPS formation by glycansucrases has been reported for lactobacilli of the species Lactobacillus reuteri, L. pontis, L. panis, L. acidophilus, and L. frumenti (28, 29). HoPS synthesis and the corresponding genes have been especially well characterized in L. reuteri (27). All EPSs used in this study were HoPSs formed in the presence of sucrose. In addition to HoPS synthesis, glycansucrases are capable of producing oligosaccharides (OSs). Several OSs are found to have prebiotic properties as they can benefit host health by acting as nondigestible food ingredients and by enhancing the growth of desired microbial members of the gastrointestinal microbiota (7). Emerging research efforts have investigated potential applications of OSs as antiadhesive agents in preventing pathogen colonization. Shoaf et al. (24) suggested that commercial oligo- and polysaccharides reduce the adherence of enteropathogenic Escherichia coli (EPEC) to cell lines. Martín-Sosa et al. (16) reported antiadhesive properties of human milk oligosaccharides against human strains of enterotoxigenic Escherichia coli (ETEC) and uropathogenic E. coli. Similarly, Martín et al. (15) confirmed the binding of milk oligosaccharides to ETEC isolated from calves. In contrast, adhesion studies with EPS are limited.The inhibition of ETEC is of special interest to the swine industry because ETEC is the primary cause for diarrhea in neo- and postnatal piglets and results in substantial economic losses. We therefore aimed to test for antiadhesive properties of EPSs synthesized by LAB and commercially available prebiotics against porcine ETEC strains. Hemagglutination assays were used; these assays are generally accepted as an effective model system for testing ETEC adherence as hemagglutination resembles the attachment of K88-positive bacteria to the gut wall (10).     

Enhancing the Production and Monosaccharide Composition of Exopolysaccharides of Lactobacillus plantarum VAL6 by Applying Thermal Stress and Increased Carbon Dioxide Concentration

Microbiology - Lactobacilli are able to produce exopolysaccharides (EPSs) with a wide diversity in structure and composition. However, changes in EPS production under environmental challenges are...     

Comparative studies of lipopolysaccharide and exopolysaccharide from a virulent strain of Pseudomonas solanacearum and from three avirulent mutants.

The composition of the Pseudomonas solanacearum lipolysaccharide (LPS) was found to be similar to that described for the LPS of enterobacteria. The lipid A contained fatty acids and glucosamine in a molar ratio of 5:2. The LPS fraction contained 2-keto-3-deoxyoctulosonic acid, L-glycero-D-mannoheptose, hexoses (glucose, rhamnose, and glucosamine), and a pentose (xylose). The LPSs from the wild-type strain (GMI1000), from the spontaneous rough mutant (GMI2000), and from their respective acridine orange-resistant (Acrr) mutants (GMI1178 and GMI2179) contained the same component sugars in their polysaccharide moieties, but the relative amounts of each sugar varied greatly. Spontaneous mutation to the rough type was characterized by a decrease in the ratio of rhamnose to glucose, whereas a reverse effect was seen for the acridine orange resistance mutation from the parent strains (GMI1000 and GMI2000) to the respective mutant strains (GMI1178 and GMI2179). The exopolysaccharide (EPS) from GMI1000 was found to be composed of two fractions: a heteropolysaccharide (galactosamine, glucose, and rhamnose) excluded from Sephadex G-50 and an additional glucan with a lower molecular weight. Strains GMI1000 and GMI1178 produced comparable amounts of EPS, GMI2179 synthesized less EPS, and GMI2000 produced no detectable EPS. High-pressure liquid chromatography and 13C nuclear magnetic resonance analyses revealed some differences between these EPSs. The glucan fraction seemed to be the major component of the EPS from GMI2179, whereas GMI1000 and GMI1178 EPSs contained both fractions and appeared to differ in the structures of their heteropolysaccharide fractions. Viscosity measurements confirmed differences between whole EPSs produced by the three strains.     

Diverse Exopolysaccharide Producing Bacteria Isolated from Milled Sugarcane: Implications for Cane Spoilage and Sucrose Yield

Bacterial deterioration of sugarcane during harvesting and processing is correlated with significant loss of sucrose yield and the accumulation of bacterial polysaccharides. Dextran, a homoglucan produced by Leuconostoc mesenteroides, has been cited as the primary polysaccharide associated with sugarcane deterioration. A culture-based approach was used to isolate extracellular polysaccharide (EPS) producing bacterial strains from milled sugarcane stalks. Ribosomal RNA sequencing analysis grouped 25 isolates into 4 genera. This study identified 2 bacterial genera not previously associated with EPS production or sucrose degradation. All isolates produced polysaccharide when grown in the presence of sucrose. Monosaccharide analysis of purified polymers by Gas Chromatography revealed 17 EPSs consisting solely of glucose (homoglucans), while the remainder contained traces of mannose or fructose. Dextranase treatment of polysaccharides yielded full digestion profiles for only 11 extracts. Incomplete hydrolysis profiles of the remaining polysaccharides suggest the release of longer oligosaccharides which may interfere with sucrose crystal formation.     

Neutral sugar composition of extracellular polysaccharides produced by strains of Butyrivibrio fibrisolvens

The extracellular polysaccharides (EPSs) produced by 37 isolates presently classified as Butyrivibrio species (or more specifically as Butyrivibrio fibrisolvens) were purified from glucose-grown cultures. The neutral sugar compositions of these EPSs were determined by both thin-layer and gas-liquid chromatographic techniques. Results showed that while the neutral sugar composition of the EPS was constant for a given strain, it varied considerably between strains. In addition, several acidic components in the EPS, of both known and unknown structure, were detected artifactually as acetylated lactones, the acetylated alditols derived from these lactone(s), or both. Two novel components, L-altrose and the acidic sugar 4-O-[1-carboxyethyl]-D-galactose, were common constituents of the EPS from some strains of B. fibrisolvens. These and other EPS compositional features were used to sort isolates of B. fibrisolvens into groups which may have taxonomic significance. A scheme for sorting isolates into these groups, and the relative relationships between groups, is proposed.