gamma-Linolenic acid production by solid-state fermentation of Mucorales strains on cereals.

Oleaginous fungi of the genus Mucorales were screened for gamma-linolenic acid (GLA) production on solid substrates containing moistened cereals. Cunninghamella elegans CCF 1318 produced the highest yields of GLA when cultivated on barley. Substrate moisture and cultivation temperature proved critical for effective GLA production. Vegetable oil supplied to the cultures improved GLA production. Rotating bottles and plastic bags were used as cultivation vessels to reproduce the conditions found in rotating drums and tray bioreactors, respectively. After 11 days of cultivation at 21 degrees C, C. elegans produced 14.2 mg of GLA per gram of dry substrate, composed of a mixture of barley, spent malt grains (SMG) and peanut oil. GLA represented 15.6% of the total fatty acids in the lipid extract.

Urea biosensor based on amperometric pH-sensing with hematein as a pH-sensitive redox mediator.

The natural dye hematein in water solution was used as a pH-sensitive redox-active mediator for amperometric pH-sensing. The electrochemical characteristics were studied using cyclic voltammetry and chronoamperometry. Several types of urea biosensors were constructed with urease on the surface of platinum and graphite composite electrodes or in the bulk of the graphite composite. They were used for the amperometric urea determination at a working potential of 0 mV (versus SCE) using 0.5 mM hematein. Detection limits and response linearity was in the micromolar range depending on the biosensor type, concentration and pH of buffers used. An interference study of various cations, anions, and substances, which may be present in real samples demonstrated good selectivity for the determination of urea. The biosensors showed good operational (>3 h) and storage (>3 months) stability. The results of urea determination in blood and urine obtained by biosensor correlated well with those obtained by a spectrophotometric reference method.

Selective and sensitive biosensor for theophylline based on xanthine oxidase electrode.

Milk and microbial xanthine oxidases (XOs) were used for the construction of amperometric enzyme electrodes. Substrate specificity differences of these enzymes were studied. Of the two enzymes, only the microbial XO was found to oxidize theophylline, but not theobromine and caffeine. The substrate specificity of microbial XO was affected by pH, where the optimum for xanthine was 5.5, while for theophylline it was in the range from 6.5 to 8.5. The theophylline biosensor showed a low detection limit of 2 x 10(-7) M and signal linearity up to 5 x 10(-5) M. The sensitivity of the microbial XO electrode to theophylline could be selectively eliminated by immersion in alkaline phosphate solution, thus allowing for the construction of a blank electrode for differential measurements. The feasibility of this approach has been demonstrated by the determination of free (unbound) and total theophylline in blood samples. The biosensor exhibited good operational (>6 h) and shelf (>3 months) stability when trehalose was used as a stabilizer of the biocatalytic layer.

Production of polyunsaturated fatty acids by Pythium ultimum in solid-state cultivation.

The oleaginous fungus Pythium ultimum was cultivated on various solid substrates in order to achieve fungal oil enriched in the polyunsaturated fatty acids arachidonic acid and eicosapentaenoic acid. Cultivation parameters, such as incubation temperature and time, substrate composition, and moisture were optimized, so as to obtain as much as 3.5 mg eicosapentaenoic acid and 2.6 mg arachidonic acid/g of wet substrate consisting of 28.5% pearled barley, 5.75% spent malt grains, 5.75% linseed oil, and 60% nutrient solution.

Succinoglycan production by solid-state fermentation with Agrobacterium tumefaciens.

Succinoglycan was produced by cultivating Agrobacterium tumefaciens on various solid substrates, including agar medium, spent malt grains, ivory nut shavings, and grated carrots, impregnated with a nutrient+ solution. Fermentations were performed on a laboratory scale, both under static conditions and with agitation, using bottles and a prototype horizontal bioreactor. Several fermentation parameters were examined and optimized, including carbon and nitrogen composition, water content and layer thickness of the substrate. The yields and rheological properties of the polymers obtained under different fermentation conditions were compared. The highest succinoglycan yield was achieved in static cultivation, reaching 42 g/l of impregnating solution, corresponding to 30 g/kg of wet substrate. The polymer production in the horizontal bioreactor was faster, but the final yield was lower (29 g/l of impregnating solution).

Simultaneous production and purification of Bacillus subtilis alpha-amylase.

Production of alpha-amylase with B. subtilis CCM 2722 in an aqueous two-phase polyethylene glycol/dextran system integrated with product purification by affinity chromatography on crosslinked starch during cultivation was studied. The medium was drawn from the bioreactor to the external settler during fermentation. After phase separation in the settler the dextran-rich bottom phase with cells was returned to the bioreactor. The PEG-rich top phase was pumped to the column with crosslinked starch and returned to the bioreactor after alpha-amylase adsorption. The same volumetric productivities, 0.53 U/mL/h, were reached in both batch and described process, but total productivity of the latter method was much higher owing to shortening upstream and downstream processing time. The enzyme of 98% homogenity in 95% yield was obtained after its elution from the column.