High bioreactor production and emulsifying activity of an unusual exopolymer by Chromohalobacter canadensis 28.

Unusual composition of an exopolymer (EP) from an obligate halophilic bacterium Chromohalobacter canadensis 28 has triggered an interest in development of an effective bioreactor process for its production. Its synthesis was investigated in 2-L bioreactor at agitation speeds at interval 600-1000 rpm, at a constant air flow rate of 0.5 vvm; aeration rates of 0.5, 1.0, and 1.5 vvm were tested at constant agitation rate of 900 rpm. EP production was affected by both, agitation and aeration. As a result twofold increase of EP yield was observed and additionally increased up to 3.08 mg/mL in a presence of surfactants. For effective scale-up of bioreactors mass transfer parameters were estimated and lowest values of KLa obtained for the highest productivity fermentation was established. Emulsification activity of EP exceeded that of trade hydrocolloids xanthan, guar gum, and cellulose. A good synergism between EP and commercial cellulose proved its potential exploration as an enhancer of emulsifying properties of trade emulsions. A pronounced lipophilic effect of EP was established toward olive oil and liquid paraffin. Cultivation of human keratinocyte cells (HaCaT) with crude EP and purified γ-polyglutamic acid (PGA) showed higher viability than control group.

Extracellular polymer substance synthesized by a halophilic bacterium Chromohalobacter canadensis 28.

Halophilic microorganisms are producers of a lot of new compounds whose properties suggest promising perspectives for their biotechnological exploration. Moderate halophilic bacterium Chromohalobacter canadensis 28 was isolated from Pomorie salterns as an extracellular polymer substance (EP) producer. The best carbon source for extracellular polymer production was found to be lactose, a sugar received as a by-product from the dairy industry. After optimization of the culture medium and physicochemical conditions for cultivation, polymer biosynthesis increased more than 2-fold. The highest level of extracellular polymer synthesis by C. canadensis 28 was observed in an unusually high NaCl concentration (15% w/v). Chemical analysis of the purified polymer revealed the presence of an exopolysaccharide (EPS) fraction (14.3% w/w) and protein fraction (72% w/w). HPLC analysis of the protein fraction showed the main presence of polyglutamic acid (PGA) (75.7% w/w). EPS fraction analysis revealed the following sugar composition (% w/w): glucosamine 36.7, glucose 32.3, rhamnose 25.4, xylose 1.7, and not identified sugar 3.9. The hydrogel formed by PGA and EPS fractions showed high swelling behavior, very good emulsifying and stabilizing properties, and good foaming ability. This is the first report for halophilic bacterium able to synthesize a polymer containing PGA fraction. The synthesized biopolymer shows an extremely high hydrophilicity, due to the simultaneous presence of PGA and EPS. The analysis of its functional properties and the presence of glucosamine in the highest proportion in EPS fraction clearly determine the potential of EP synthesized by C. canadensis 28 for application in the cosmetics industry.

Production and properties of two novel exopolysaccharides synthesized by a thermophilic bacterium Aeribacillus pallidus 418.

Synthesis of innovative exocellular polysaccharides (EPSs) was reported for few thermophilic microorganisms as one of the mechanisms for surviving at high temperature. Thermophilic aerobic spore-forming bacteria able to produce exopolysaccharides were isolated from hydrothermal springs in Bulgaria. They were referred to four species, such as Aeribacillus pallidus, Geobacillus toebii, Brevibacillus thermoruber, and Anoxybacillus kestanbolensis. The highest production was established for the strain 418, whose phylogenetic and phenotypic properties referred it to the species A. pallidus. Maltose and NH4Cl were observed to be correspondingly the best carbon and nitrogen sources and production yield was increased more than twofold in the process of culture condition optimization. After purification of the polymer fraction, a presence of two different EPSs, electroneutral EPS 1 and negatively charged EPS 2, in a relative weight ratio 3:2.2 was established. They were heteropolysaccharides consisting of unusual high variety of sugars (six for EPS 1 and seven for EPS 2). Six of the sugars were common for both EPSs. The main sugar in EPS 1 was mannose (69.3 %); smaller quantities of glucose (11.2 %), galactosamine (6.3 %), glucosamine (5.4 %), galactose (4.7 %), and ribose (2.9 %) were also identified. The main sugar in EPS 2 was also mannose (33.9 %), followed by galactose (17.9 %), glucose (15.5 %), galactosamine (11.7 %), glucosamine (8.1 %), ribose (5.3 %), and arabinose (4.9 %). Both polymers showed high molecular weight and high thermostability.

Submerged culture process for biomass and exopolysaccharide production by Antarctic yeast: some engineering considerations.

Production of biomass and extracellular polysaccharide (EPS) from psychrophilic Sporobolomyces salmonicolor AL1 in a stirred bioreactor was studied. The aspects of production technical-scale parameters, namely, bioreactor flow field, biomass and EPS production rates, oxygen mass transfer per input power, as well as important product properties, such as rheology and stability of EPS mixtures, were considered. The bioprocess was found to proceed in non-Newtonian flow with consistency coefficient rising typically to 0.03 Pa.s(n) and flow index declining to 0.7. Flow modeling was carried out and showed good homogenization for substrate delivery at agitation rates exceeding 400 rpm. Agitation rates lower than 400 rpm were considered counterproductive due to flow field non-uniformity. The cell density reached 5 g/l and EPS production yield reached 5.5 g/l at production rate 0.057 g EPS/l per hour (0.01 g EPS/g biomass per hour). Oxygen uptake rate and oxygen transfer rate were in the range of 0.5-1.7 mmolO2/l per hour and 2-4.7 mmolO2/l per hour, respectively. The mass transfer coefficient at reaction conditions was found to be in the range [Formula: see text]. The bioprocess biological performance was higher at moderate agitation speed and revealed biomass diminution and cell inactivation by increasing impeller revolutions and shear rate. The product EPS was found to introduce shear-thinning behavior in water solutions with apparent viscosity of up to 30 mPa.s and to stabilize 1-2 % oil-in-water emulsions improving their lipophilic properties. The emulsion dispersion index was found to be comparable with the one of Arlacel 165, the emulsifier used in cosmetic. The long-term performance of the complex cream mixtures of the glucomannan prepared in commercial format was found promising for further application.

Optimization of rosmarinic acid production by Lavandula vera MM plant cell suspension in a laboratory bioreactor.

The all-round effect of dissolved oxygen concentration, agitation speed, and temperature on the rosmarinic acid production by Lavandula vera MM cell suspension was studied in a 3-L laboratory bioreactor by means of the modified Simplex method. Polynomial regression models were elaborated for description of the process of rosmarinic acid production (Y) in the bioreactor as a consequence of the variation of the dissolved oxygen (X(1)) concentration between 10% and 50%; agitation (X(2)) between 100 and 400 rpm; and temperature (X(3)) between 22 and 30 degrees C. The optimization made it possible to establish the optimal conditions for the biosynthesis of rosmarinic acid by L. vera MM: dissolved oxygen (X(1)*), 50% of air saturation; agitation (X(2)*), 400 rpm; and temperature (X(3)*), 29.9 degrees C, where maximal yield (Y(max)) of 3489.4 mg/L of rosmarinic acid was achieved (2 times higher compared with the shake-flasks cultivation).

Structural development of sucrose-sweetened and sucrose-free sponge cakes during baking.

The influence of sucrose, wheat starch and sorbitol upon the heat- and mass-exchanging processes forming the structure of sponge cake was studied. Under the influence of wheat starch and sorbitol the structure of the sucrose-free sponge cake was formed at more uniform total moisture release. This process was done at lower temperatures and smoother change of the sponge cake height with respect to the sucrose-sweetened sponge cake. The porous and steady structure of both cakes was finally formed at identical time--between 18th and 19th minute, at the applied conditions for baking of each batter (metal pan with diameter 15.4 cm and depth 6.2 cm containing 300 g of batter and placed in an electric oven "Rahovetz-02", Bulgaria for 30 min at 180 degrees C). The water-losses at the end of baking (10.30% and 10.40% for the sucrose-sweetened cake and sucrose-free cake, respectively) and the final temperatures reached in the crumb central layers (96.6 degrees C and 96.3 degrees C for the sucrose-sweetened cake and sucrose-free cake, respectively) during baking of both samples were not statistically different. The addition of wheat starch and sorbitol in sucrose-free sponge cake lead to the statistically different values for the porosity (76.15% and 72.98%) and the volume (1014.17 cm3 and 984.25 cm3) of the sucrose-sweetened and sucrose-free sponge cakes, respectively. As a result, the sucrose-free sponge cake formed during baking had a more homogeneous and finer microstructure with respect to that ofthe sucrose-sweetened one.