Glycogen Metabolism in Aureobasidium Pullulans: A Glycogen Synthetase with Unusual Activation Properties

Abstract

Cultures of filamentous fungi that secrete significant amounts of exopolysaccharides are among the most difficult of fermentation fluids, presenting difficulties in the areas of aeration, agitation, mixing, and control that may in turn impact the physiology of the microorganism in an undesirable manner. The fungus Sclerotium glucanicum, which produces a potentially useful exopolysaccharide known as scleroglucan, illustrates many such difficulties. This review discusses in detail the range of physiological studies on the producing microorganism itself, including those concerning formation of “undesirable” byproducts, principally oxalate, but also, under certain conditions, other TCA cycle acids. In addition, the bioreactor technology in use for production of this type of biopolymer is discussed in relation to the difficulties such fluid types present. The potential of pneumatically agitated reactors for such production is evaluated, and the lack of fundamental studies on such reactors and on the hydrodynamics and mixing behavior of such complex fluids is pointed out.     

Rhizobium extracellular structures in the symbiosis

The extracellular and surface polysaccharides produced by Rhizobium species constitute a composite macromolecular interface between the bacterial cell and its environment. Several of these polysaccharides are involved in the complex series of interactions leading to the establishment of an effective Rhizobium-legume symbiosis. Extracellular heteropolysaccharides (EPSs) are found in culture supernatants, while capsular polysaccharides adhere to the cell surface. Cyclic (1–2)--d glucan is a periplasmic oligosaccharide that has also been found in the culture supernatants of some strains. The lipopolysaccharides (LPSs), which form part of the outer membrane and contain the O-somatic antigens, comprise the other major group of extracellular polysaccharides. In this review we will describe the major Rhizobium extracellular structures and their role in symbiosis with leguminous plants.The authors are with the Departamento de Microbiologia y Parasitologia, Facultad de Farmacia. Universidad de Sevilla, 41012 Sevilla, Spain     

Damages Caused by Irradiation in the Presence of NaCl in Macromolecular Components of E. coli

Damages in macromolecular components and in their functions of E. coli produced by gamma irradiation in the presence of higher concentration of NaCl were determined with reference to the basic mechanism for NaCl radiosensitization. As the results, inhibitions of DNA repair, of DNA degradation, of macromolecular syntheses and of RNA species formation were found in DNA repair proficient strains. The number of radiation-induced DNA-strand breaks was not increased by the presence of 1 m NaCl during irradiation. It was suggested that inhibition of DNA repair may be of primary importance among these lesions for NaCl radiosensitization.     

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.     

Role of plant growth-promoting rhizobacteria and their exopolysaccharide in drought tolerance of maize

The present study deals with the isolation and characterization of exopolysaccharides (EPS) produced by the plant growth-promoting rhizobacteria (PGPR) from arid and semiarid regions of Pakistan, and to investigate the drought tolerance potential of these PGPR on maize when used as bioinoculant alone and in combination with their respective EPS. Three bacterial strains Proteus penneri (Pp1), Pseudomonas aeruginosa (Pa2), and Alcaligenes faecalis (AF3) were selected as EPS-producing bacteria on the basis of mucoid colony formation. All these strains were gram negative, motile, and positive for catalase. Strain Pp1 was positive for oxidase test and was phosphate solubilizing, while Pa2 and AF3 were negative. The isolated strains were sequenced using 16SrRNA. Total soluble sugar, protein, uronic acid, emulsification activity, and Fourier-transformed infrared spectroscopy of EPS were determined. Drought stress had significant adverse effects on growth of maize seedlings. Seed bacterization of maize with EPS-producing bacterial strains in combination with their respective EPS improved soil moisture contents, plant biomass, root and shoot length, and leaf area. Under drought stress, the inoculated plants showed increase in relative water content, protein, and sugar though the proline content and the activities of antioxidant enzymes were decreased. The Pa2 strain isolated from semiarid region was most potent PGPR under drought stress. Consortia of inocula and their respective EPS showed greater potential to drought tolerance compared to PGPR inocula used alone.     

细虫草胞外多糖对小鼠腹腔巨噬细胞免疫功能研究

本实验在体外条件下,以人工发酵培养的细虫草胞外多糖OgE、OgE-F1和OgE-F2作用于小鼠腹腔巨噬细胞RAW264.7,通过测定其对巨噬细胞的增殖率、代谢MTT活力、NO分泌和吞噬能力的影响,评价细虫草胞外多糖的免疫调节活性。结果表明,细虫草多糖对巨噬细胞无细胞毒性,且能促进巨噬细胞代谢MTT活力;在0.2mg/mL^1.0mg/mL浓度范围内,多糖呈剂量依赖性的促进巨噬细胞分泌NO水平和吞噬能力。本研究表明,细虫草多糖能有效地增强小鼠巨噬细胞的活性,潜在地可改善小鼠的先天性免疫调节。     

灵芝不同菌株胞外多糖的单糖组成和抗氧化活性分析

灵芝多糖是药用真菌灵芝的主要活性成分之一。早期研究发现不同灵芝子实体多糖的单糖组成和活性存在差异,但不同灵芝菌株胞外多糖的单糖组成和活性是否有区别仍不清楚。本研究通过液体发酵获得灵芝菌株5.26和5.616的胞外多糖,使用DEAE-cellulose和Sephadex G-200柱色谱分离纯化得到了两种多糖(5.26-2-1和5.616-2-1),并对5.26-2-1,5.616-2-1的单糖组成和抗氧化活性进行了分析。结果表明,5.26-2-1主要由甘露糖、半乳糖醛酸、半乳糖和葡萄糖组成,而5.616-2-1主要由甘露糖、半乳糖和葡萄糖组成。5.26-2-1中葡萄糖的摩尔百分比为63.97%,显著高于5.616-2-1 (29.3%),但半乳糖的摩尔百分比为9.34%,显著低于5.616-2-1 (42.78%)。当多糖质量浓度为2 mg/mL时,5.26-2-1的Fe²⁺螯合能力、对1,1-二苯基-2-三硝基苯肼(DPPH)和羟自由基(·OH)的最大清除率分别为71.9%、71.3%和60.8%,显著高于5.616-2-1的值(63.5%、60.4%和51.8%)。本研究有助于灵芝多糖的开发与利用,为进一步探究灵芝多糖的构效关系提供了重要信息。     

Involvement of a plasmid in exopolysaccharide production by Klebsiella oxytoca

A Klebsiella oxytoca isolate which can produce significant levels of an exopolysaccharide using whey as a growth substrate has been reported. The plasmid profile of this isolate was shown to be different from that of the non-exopolysaccharide-producing K. oxytoca ATCC 43863. Irreversible curing of the single plasmid in the K. oxytoca isolate was achieved using 30 g acriflavin/ml. The ability to produce the exopolysaccharide was lost with the curing of the plasmid (parent strain produced a medium viscosity of 1260 cP at 1 s^–1 compared to 1.6 cP at 200 s^–1 produced by the cured strain). However, the ability to metabolize lactose was not significantly affected by curing, and both the parent and the cured strain produced similar levels of viable cells (~10⁹ cfu/ml) after 62 h growth on lactose-rich medium. The exopolysaccharide-producing ability of the isolate was stable for at least 139 generations.     

Production of slime polysaccharide by EHEC and STEC as well as the influence of culture conditions on slime production in Escherichia coli O157:H7

AIMS: To quantify the slime polysaccharide, composed of colanic acid (CA), produced by enterohaemorrhagic and Shiga toxin-producing Escherichia coli (EHEC and STEC) and to determine the influence of culture conditions on CA production in E. coli O157:H7. METHODS AND RESULTS: The study examined the amounts of CA produced by EHEC and STEC, and evaluated the production of CA in E. coli O157:H7 as influenced by medium pH and incubation temperatures. The results indicated that the amounts of CA produced by EHEC and STEC vary to a great extent and CA production in E. coli O157:H7 is influenced by the tested culture conditions. CONCLUSIONS: The abilities of EHEC and STEC to produce CA differ. Medium pH and incubation temperature are among the important factors affecting CA production in E. coli O157:H7. SIGNIFICANCE AND IMPACT OF THE STUDY: Slime polysaccharide can affect the abilities of E. coli O157:H7 cells to combat environmental stress. This study contributes to a better understanding of the physiological factors influencing slime polysaccharide production in EHEC and STEC.     

Optimization of submerged culture requirements for the production of mycelial growth and exopolysaccharide by Cordyceps jiangxiensis JXPJ 0109

AIMS: The objective of the present study was to investigate the optimal culture requirements for mycelial growth and exopolysaccharide production by Cordyceps jiangxiensis JXPJ 0109 in submerged culture. METHODS AND RESULTS: The effects of medium ingredients (i.e. carbon and nitrogen sources, and growth factor) and other culture requirements (i.e. initial pH, temperature, etc.) on the production of mycelia and exopolysaccharide were observed using a one-factor-at-a-time method. More suitable culture requirements for mycelial growth and exopolysaccharide production were proved to be maltose, glycerol, tryptone, soya bean steep powder, yeast extract, medium capacity 200 ml in a 500-ml flask, agitation rate 180 rev min(-1), seed age 4-8 days, inoculum size 2.5-7.5% (v/v), etc. The optimal temperatures and initial pHs for mycelial growth and exopolysaccharide production were at 26 degrees C and pH 5 and at 28 degrees C and pH 7, respectively, and corresponding optimal culture age were observed to be 8 and 10 days respectively. According to the primary results of the one-factor-at-a-time experiments, the optimal medium for the mycelial growth and exopolysaccharide production were obtained using an orthogonal layout method to optimize further. Herein the effects of medium ingredients on the mycelial growth of C. jiangxiensis JXPJ 0109 were in the order of yeast extract > tryptone > maltose > CaCl2 > glycerol > MgSO4 > KH2PO4 and the optimal concentration of each composition was 15 g maltose (food-grade), 10 g glycerol, 10 g tryptone, 10 g yeast extract, 1 g KH2PO4, 0.2 g MgSO4, and 0.5 g CaCl2 in 1 l of distilled water, while the order of effects of those components on exopolysaccharide production was yeast extract > maltose > tryptone > glycerol > KH2PO4 > CaCl2 > MgSO4, corresponding to the optimal concentration of medium was as follows: 20 g maltose (food-grade), 8 g glycerol, 5 g tryptone, 10 g yeast extract, 1 g KH2PO4, and 0.5 g CaCl2 in 1 l of distilled water. CONCLUSIONS: Under the optimal culture requirements, the maximum exopolysaccharide production reached 3.5 g l(-1) after 10 days of fermentation, while the maximum production of mycelial growth achieved 14.5 g l(-1) after 8 days of fermentation. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report on the submerged culture requirements for mycelial growth and exopolysaccharide in C. jiangxiensis, and this two-step optimization strategy in this study can be widely applied to other microbial fermentation processes.