Abstract: Bioconversion of tannin to gallic acid from powder of teri pod (Caesalpinia digyna) cover was achieved by the locally isolated fungus, Rhizopus oryzae, in a bioreactor with a perforated float for carrying solid substrate and induced inoculum. Modified Czapek-Dox medium, put beneath the perforated float, with 2% tannic acid at pH 4.5, temperature 32°C, 93% relative humidity, incubated for 3 days with 3-day-old inoculum was optimum for the synthesis of tannase vis-a-vis gallic acid production. Conversion of tannin to gallic acid was 90.9%. Diethyl ether was used as the solvent for extraction of gallic acid from the fermented biomass.

Abstract: D.B. Wilson, D.C. Irwin: Genetics and Properties of Cellulases.- N.S. Mosier, P. Hall, C.M. Ladisch, M.R. Ladisch: Reaction Kinetics, Molecular Action, and Mechanisms of Cellulolytic Proteins.- J.S. Tolan, B. Foody: Cellulase from Submerged Fermentation.- P. Cen, L. Xia: Production of Cellulase by Solid-State Fermentation.- Y.Y. Lee, P. Iyer, R.W. Torget: Dilute-Acid Hydrolysis of Lignocellulosic Biomass.- T.W. Jeffries, N.-Q. Shi: Genetic Engineering for Improved Xylose Fermentation by Yeasts.- N.W.Y. Ho, Z. Chen, A.P. Brainard, M. Sedlak: Successful Design and Development of Genetically Engineered Saccharomyces Yeasts for Effective Cofermentation of Glucose and Xylose from Cellulosic Biomass to Fuel Ethanol.- H.M. Muhlemann, H.R. Bungay: Research Perspectives for Bioconversion of Scrap Paper. C.S. Gong, N.J. Cao, J. Du, G.T. Tsao: Ethanol Production from Renewable Resources.- G.T. Tsao, N.J. Cao, J. Du, C.S. Gong: Production of Multifunctional Organic Acids from Renewable Resources.

Abstract: We describe the biocatalytic production of 3-phenylcatechol from 2-phenylphenol with the whole cell biocatalyst Escherichia coli JM101 (pHBP461). The recombinant produces 2-hydroxybiphenyl 3-monooxygenase, an enzyme from Pseudomonas azelaica HBP1. This enzyme introduces a hydroxyl-group at the C3-position of a variety of 2-substituted phenols, such as 2-phenylphenol. This permits the biocatalytic production of 3-substituted catechols, which are difficult to synthesize chemically. Both 2-phenylphenol and 3-phenylcatechol are highly toxic to E. coli. The toxic effects of 2-phenylphenol were minimized by feeding this substrate to the reactor at a rate slightly below the maximum biooxidation rate. As a result, the substrate concentration in the reactor remained below toxic levels during the bioconversion. The toxic product formed was removed by continuous adsorption on the solid resin Amberlite XAD-4. To this end the reaction mixture, containing the biocatalyst, was pumped continuously through an external loop with a fluidized bed of the resin. This resin efficiently and quantitatively adsorbed both 3-phenylcatechol and the remaining trace amounts of 2-phenylphenol. Consequently, the concentrations of these compounds were kept at subtoxic levels (below 100 mg L-1) and gram amounts of 3-phenylcatechol were produced with space-time yields of up to 0.39 g L-1 h-1. The product was recovered from the resin by acidic methanol elution and purified by recrystallization from n-hexane resulting in overall yields exceeding 59%. The optimized system served as a surprisingly simple and efficient integrated process, that allows the bioconversion of toxic substrates to toxic products with whole cell biocatalysts.

Abstract: The effects of the steam-explosion treatment on aqueous fractionation and bioconversion of wheat straw have been investigated. The treatments have been carried out in batch and continuous reactors with capacity of 0.5 Kg/cycle and 150 Kg/h, respectively. The exploded materials have been sequentially extracted with water at 65°C and sodium hydroxide 1.5%. Analytical determinations of liquid fractions and solid residues have led to the fractionation pattern of the carbohydrates as monomers, oligomers, and polymers. Evaluations of total acidity, ash content, and lignin recovery have improved understanding of the process. This part of the work has allowed us to derive: the empiric relationship between the batch and the continuous reactors and the yield and availability of pentoses and hexoses in various phases. Selected samples have been tested in enzymatic-hydrolysis experiments, pointing out the effect of treatment severity and reactor used on the saccharification yield.

Abstract: gamma-Decalactone (gamma-C10) is known to be highly toxic for the microorganims used for its production In this work, three techniques were studied in order to overcome this toxicity during a bioconversion process using ricinoleic acid as precursor of the lactone: in situ trapping in oily phases, in porous hydrophobic sorbents and in beta-cyclodextrins Oily phases added to the media (olive, Miglyol, tributyrin, and paraffin) had a protective effect on Sp salmonicolor, and they improved the lactone production beta-cyclodextrins, which have a hydrophobic cavity that can trap molecules such as gamma-C10, have been used to protect the yeasts The results showed insufficient preservation of cell viability Some sorbents (activated carbon and polystyrene-based sorbents) were successfully tested during bioconversion In all cases viability exceeded the reference one Nevertheless the aroma production was 30% lower than the reference All of these solutions led to some enhancement of the cell viability during bioconversion of methyl ricinoleate to gamma-C10 For improvement of the lactone production, the oil trapping method seemed to be the best with the experimental conditions tested

Abstract: High-density cultures of Pycnoporus cinnabarinus were tested with a view to optimisation of ferulic acid bioconversion into vanillin. The dry weight was increased fourfold by using glucose, fructose or a mixture of glucose and phospholipids as carbon source instead of maltose, the carbon source previously used. 5 mmol l−1 vanillin, i.e. 760 mg l−1, was produced over 15 days with glucose-phospholipid medium. In contrast, formation of vanillin was lower using glucose or fructose compared to the maltose control. A bioreactor (2 l) with a glucose-phospholipid medium gave a molar yield of vanillin of 61% (4 mmol l−1). An alternative strategy was to grow the fungus on a glucose or fructose medium for 3 days, then switch to maltose during the bioconversion phase: this method allowed 3.3 mmol l−1 vanillin to be obtained in 10 days. Many by-products such as methoxyhydroquinone and vanillyl alcohol were also produced.

Abstract: In this research study fungi isolated from effluent of ethanol factories were identified. Optimal conditions for single cell protein (SCP) production and COD reduction of sugar beet stillage is specified for a species of Hansenula in continuous culture. Under these conditions 5.7 g dm−3 biomass was produced and 31% of COD was reduced without addition of further nutrients in beet molasses stillage. By addition of nitrogen and phosphorus sources, biomass production and COD reduction increased to 8.5 g dm−3 and 35.7%, respectively. The crude protein content of SCP in the absence of additives was 39.6% but this was increased to 50.6% in the presence of additives. The amount of essential amino acids measured was greater than that of the reference and are comparable with some other food proteins, such as soya bean and fish meal. The final effluent of SCP production was recycled to fermentation stage. The results represent 70% reduction of stillage volume.

Abstract: At present, there is little information on the optimization of the degradation of polycyclic aromatic hydrocarbons (PAH) by deuteromycete filamentous fungi, a reaction catalyzed by cytochrome P450 monooxygenases. We utilized response-surface methodology to determine the optimal growth conditions for the oxidation of the PAH pyrene by Penicillium janthinellum SFU403, with respect to the variables glucose concentration, nitrate concentration and bioconversion time. Models were derived for the relationship between the variables tested and the level of the pyrene oxidation products, 1-pyrenol (1-PY) and pyrenequinones (PQ). Production of 1-PY and PQ were optimized by the same glucose and nitrate concentrations: 2.5% glucose and 1.5% sodium nitrate. The optimized 1-PY and PQ bioconversion times were 71 h and 73 h respectively. These conditions improved the yield of 1-PY by fivefold and PQ were more than 100-fold higher than the baseline levels obtained in this study. The optimized PQ yield represented 95% of the initial pyrene, thus the total optimised pyrene bioconversion to 1-PY and PQ was approximately 100%. Concentrations of glucose exceeding 4.0% repressed pyrene hydroxylation. Pyrene hydroxylation occurred almost exclusively during the deceleration phase of culture growth.

Abstract: A mixed solids waste (MSW) feedstock, comprising construction lumber waste (35% oven-dry basis), almond tree prunings (20%), wheat straw (20%), office waste paper (12.5%), and newsprint (12.5%), was converted to ethanol via dilute-acid pretreatment followed by enzymatic hydrolysis and yeast fermentation. The MSW was pretreated with dilute sulfuric acid (0.4% w/w) at 210°C for 3 min in a 4-L steam explosion reactor, then washed with water to recover the solubilized hemicellulose. The digestibility of water-washed, pretreated MSW was 90% in batch enzymatic hydrolysis at 66 FPU/g cellulose. Using an enzyme-recycle bioreactor system, greater than 90% cellulose hydrolysis was achieved at a net enzyme loading of about 10 FPU/g cellulose. Enzyme recycling using membrane filtration and a fed-batch fermentation technique is a promising option for significantly reducing the cost of enzyme in cellulose hydrolysis. The hexose sugars were readily fermentable using a Saccharomyces cerevisiae yeast strain that was adapted to the hydrolysate. Solid residue after enzyme digestion was subjected to various furnace experiments designed to assess the fouling and slagging characteristics. Results of these analyses suggest the residue to be of a low to moderate slagging and fouling type if burned by itself.

Abstract: The production of low molecular weight esters as flavor compounds by biotechnological processes has a potential interest for the food industry. The use of natural available substrates and enzymes is an essential part of the process design, because the products may obtain natural label. In this study, direct esterification of citronellol and geraniol with short-chain fatty acids catalyzed by free lipase from Mucor miehei was performed with high yields in n-hexane. The effects of the acid:alcohol ratio on the bioconversion rate of increasing chain length esters was investigated. To reach the optimum yield, substrates and enzyme concentration were determined. The inhibiting effects of acid are strongly attenuated by reducing the quantity of acid and increasing the amount of enzyme in media following the optimum values. Improvements have been made to increase the ester purity. The consumption of excess substrate by adding calculated amounts of acid gives a 10% yield enhancement, and leads to 100% pure terpenyl esters. The first steps to a scale-up application were attempted using a reactor that allowed us to produce ester quantities up to 100 cm3. Separation and purification of the products were treated with success, underlining the lipase stability and efficiency under the conditions of this study. The ability to recover the enzyme, and reusing it in bioconversions, plays a major role in reducing the cost of the overall process.

Abstract: Cellulose-rich residue was obtained by a pectinase treatment of sugar beet pulp and was enzymatically degraded to produce cellobiose. From sugar beet pulp, the hydrolysis yielded 5.3% of cellobiose and 3.8% of glucose. The cellobiose-rich fraction was added in a culture medium of the filamentous fungus Pycnoporus cinnabarinus to test it as an activator of vanillin production. Adding the sugar beet-derived cellobiose-rich fraction to a 3-day-old culture yielded a 3.3-fold increase in vanillin production when compared to a culture devoid of cellobiose, while commercial cellobiose yielded a 3.1-fold increase.

Abstract: In this study, a facultative bacterium that converts fumarate to succinate at a high yield was isolated. The yield of bioconversion was enhanced about 1.2 times by addition of glucose into culture medium at an initial concentration of 6 g/L. When the initial cell density was high (2 g/L), the succinate produced at pH 7.0 for initial fumarate concentrations of 30, 50, 80, and 100 g/L were 29.3,40.9,63.6, and 82.5 g/L, respectively, showing an increase with the initial fumarate concentration. The high yield of 96.8%/mole of fumarate in just 4 h was obtained at the initial fumarate concentration of 30 g/L. Comparing these values to those obtained with low cell culture (0.2 g/L), we found that the amount of succinate produced was similar, but the production rate in the high cell culture was about three times higher than was the case in the low cell culture. This strain converted fumarate to succinate at a rate of 3.5 g/L · h under the sparge of CO2.

Abstract: Recombinant Escherichia coli cells expressing the toluene dioxygenase (TDO) genes from Pseudomonas putida convert indene to cis-1S,2R-indandiol, a potentially important intermediate for the chemical synthesis of the HIV-1 protease inhibitor, Crixivan. A bioconversion process was developed through optimization of medium composition and reaction conditions at the shake-flask and 23-1 fermentor scales. A cis-1,2-indandiol productivity of approx. 1000 mg/l was achieved with construct TDO123, which represents a 50-fold increase over the initial titer. Varying the bioconversion conditions did not change the enantiomeric excess (e.e.) for the 1S,2R enantiomer from about 30%, suggesting that toluene dioxygenase intrinsically converts indene to 1S,2R- and 1R,2S-indandiols at a ratio of 2:1. Further inclusion of the Pseudomonas dehydrogenase gene in construct D160-1 led to the production of chirally pure cis-1S,2R-indandiol (e.e. > 99%) as a result of the selective degradation of the 1R,2S enantiomer, with the overall yield (650 mg/l) proportionally reduced. A single stage process was developed for D160-1 and scaled up to the 23-1 fermentor, achieving a cis-1S,2R-indandiol titer of 1200 mg/l.

Abstract: Many pseudomonads and other bacteria can grow on aliphatic and aromatic hydrocarbons that occur in the environment. We are examining the potential of such organisms as biocatalysts for the oxidation of a variety of substituted aliphatic and aromatic compounds. In order to attain a high production rate of oxidation products via such biotransformations, we have focused on two-liquid phase culture systems. In these systems cells are grown in liquid media consisting of an aqueous phase containing water soluble growth substrates and droplets of a water immiscible organic solvent containing bioconversion substrates and products.

Abstract: Bioconversion of fumarate to succinate by Enterococcus sp. RKY1 was enhanced when Tween surfactant, organic solvent, and vegetable oil were added to the fermentation medium. The maximum amount of succinate produced was 80.4 g/l after a 24 h incubation when Tween 80 was added to the culture to a final concentration of 0.1 g/l. Triton X-100 was observed to damage the enzymes and inhibit the formation of succinate. The addition of 10 ml/l acetone increased the production of succinate by 110%. Vegetable oils used were found to be effective for succinate production as well as for the cell growth. Similar productivity increases were obtained with corn oil and Tween 80 plus biotin with the total productivity being 3.6 g/l/h, and 3.5 g/l/h, respectively, which was approximately 25% greater than that of the control. Therefore, these results indicate that corn oil can be considered the most appropriate agent for the production of succinate where succinic acid was primarily used in the production of food, medicine, and cosmetics.