Detoxification of model phenolic compounds in lignocellulosic hydrolysates with peroxidase for butanol production from Clostridium beijerinckii.

In the present study, we investigated the peroxidase-catalyzed detoxification of model phenolic compounds and evaluated the inhibitory effects of the detoxified solution on butanol production by Clostridium beijerinckii National Collection of Industrial and Marine Bacteria Ltd. 8052. The six phenolic compounds, p-coumaric acid, ferulic acid, 4-hydroxybenzoic acid, vanillic acid, syringaldehyde, and vanillin, were selected as model fermentation inhibitors generated during pretreatment and hydrolysis of lignocellulose. The enzyme reaction was optimized as a function of the reaction conditions of pH, peroxidase concentration, and hydrogen peroxide to substrate ratio. Most of the tested phenolics have a broad optimum pH range of 6.0 to 9. Removal efficiency increased with the molar ratio of H(2)O(2) to each compound up to 0.5-1.25. In the case of p-coumaric acid, ferulic acid, vanillic acid, and vanillin, the removal efficiency was almost 100% with only 0.01 microM of enzyme. The tested phenolic compounds (1 g/L) inhibited cell growth by 64-74%, while completely inhibiting the production of butanol. Although syringaldehyde and vanillin were less toxic on cell growth, the level of inhibition on the butanol production was quite different. The detoxified solution remarkably improved cell growth and surprisingly increased butanol production to the level of the control. Hence, our present study, using peroxidase for the removal of model phenolic compounds, could be applied towards the detoxification of lignocellulosic hydrolysates for butanol fermentation.

Electro-biocatalytic production of formate from carbon dioxide using an oxygen-stable whole cell biocatalyst.

The use of biocatalysts to convert CO2 into useful chemicals is a promising alternative to chemical conversion. In this study, the electro-biocatalytic conversion of CO2 to formate was attempted with a whole cell biocatalyst. Eight species of Methylobacteria were tested for CO2 reduction, and one of them, Methylobacterium extorquens AM1, exhibited an exceptionally higher capability to synthesize formate from CO2 by supplying electrons with electrodes, which produced formate concentrations of up to 60mM. The oxygen stability of the biocatalyst was investigated, and the results indicated that the whole cell catalyst still exhibited CO2 reduction activity even after being exposed to oxygen gas. From the results, we could demonstrate the electro-biocatalytic conversion of CO2 to formate using an obligate aerobe, M. extorquens AM1, as a whole cell biocatalyst without providing extra cofactors or hydrogen gas. This electro-biocatalytic process suggests a promising approach toward feasible way of CO2 conversion to formate.

Efficient CO2-reducing activity of NAD-dependent formate dehydrogenase from Thiobacillus sp. KNK65MA for formate production from CO2 gas.

NAD-dependent formate dehydrogenase (FDH) from Candida boidinii (CbFDH) has been widely used in various CO2-reduction systems but its practical applications are often impeded due to low CO2-reducing activity. In this study, we demonstrated superior CO2-reducing properties of FDH from Thiobacillus sp. KNK65MA (TsFDH) for production of formate from CO2 gas. To discover more efficient CO2-reducing FDHs than a reference enzyme, i.e. CbFDH, five FDHs were selected with biochemical properties and then, their CO2-reducing activities were evaluated. All FDHs including CbFDH showed better CO2-reducing activities at acidic pHs than at neutral pHs and four FDHs were more active than CbFDH in the CO2 reduction reaction. In particular, the FDH from Thiobacillus sp. KNK65MA (TsFDH) exhibited the highest CO2-reducing activity and had a dramatic preference for the reduction reaction, i.e., a 84.2-fold higher ratio of CO2 reduction to formate oxidation in catalytic efficiency (kcat/KB) compared to CbFDH. Formate was produced from CO2 gas using TsFDH and CbFDH, and TsFDH showed a 5.8-fold higher formate production rate than CbFDH. A sequence and structural comparison showed that FDHs with relatively high CO2-reducing activities had elongated N- and C-terminal loops. The experimental results demonstrate that TsFDH can be an alternative to CbFDH as a biocatalyst in CO2 reduction systems.

Lipase-catalyzed enantioselective synthesis of (R,R)-lactide from alkyl lactate to produce PDLA (poly D-lactic acid) and stereocomplex PLA (poly lactic acid).

R-lactide, a pivotal monomer for the production of poly (D-lactic acid) (PDLA) or stereocomplex poly (lactic acid) (PLA) was synthesized from alkyl (R)-lactate through a lipase-catalyzed reaction without racemization. From among several types of lipase, only lipase B from Candida antarctica (Novozym 435; CAL-B) was effective in the reaction that synthesized (R,R)-lactide. Enantiopure (R,R)-lactide, which consisted of over 99% enantiomeric excess, was synthesized from methyl (R)-lactate through CAL-B catalysis. Removal of the methanol by-product was critical to obtain a high level of lactide conversion. The (R,R)-lactide yield was 56% in a reaction containing 100 mg of Novozym 435, 10 mM methyl (R)-lactate and 1500 mg of molecular sieve 5A in methyl tert-butyl ether (MTBE). The important monomer (R,R)-lactide that is required for the production of the widely recognized bio-plastic PDLA and the PLA stereocomplex can be obtained using this novel synthetic method.

Ethanol production from acid hydrolysates based on the construction and demolition wood waste using Pichia stipitis.

The feasibility of ethanol production from the construction and demolition (C&D) wood waste acid hydrolysates was investigated. The chemical compositions of the classified C&D wood waste were analyzed. Concentrated sulfuric acid hydrolysis was used to obtain the saccharide hydrolysates and the inhibitors in the hydrolysates were also analyzed. The C&D wood waste composed of lumber, plywood, particleboard, and medium density fiberboard (MDF) had polysaccharide (cellulose, xylan, and glucomannan) fractions of 60.7-67.9%. The sugar composition (glucose, xylose, and mannose) of the C&D wood wastes varied according to the type of wood. The additives used in the wood processing did not appear to be released into the saccharide solution under acid hydrolysis. Although some fermentation inhibitors were detected in the hydrolysates, they did not affect the ethanol production by Pichia stipitis. The hexose sugar-based ethanol yield and ethanol yield efficiency were 0.42-0.46 g ethanol/g substrate and 84.7-90.7%, respectively. Therefore, the C&D wood wastes dumped in landfill sites could be used as a raw material feedstock for the production of bioethanol.

Enzymatic coproduction of biodiesel and glycerol carbonate from soybean oil and dimethyl carbonate.

The enzymatic coproduction of biodiesel and glycerol carbonate by the transesterification of soybean oil was studied using lipase as catalyst in organic solvent. To produce biodiesel and glycerol carbonate simultaneously, experiments were designed sequentially. Enzyme screening, the molar ratio of dimethyl carbonate (DMC) to soybean oil, reaction temperature and solvent effects were investigated. The results of enzyme screening, at 100 g/L Novozym 435 (immobilized Candida antarctica lipase B), biodiesel and glycerol carbonate showed conversions of 58.7% and 50.7%, respectively. The optimal conditions were 60 °C, 100 g/L Novozym 435, 6.0:1 molar ratio with tert-butanol as solvent: 84.9% biodiesel and 92.0% glycerol carbonate production was achieved.

Enhanced ethanol production from deacetylated yellow poplar acid hydrolysate by Pichia stipitis.

In this study, alkaline-pretreatment for the extraction of acetic acid from xylan of hemicellulose was introduced prior to concentrated acid hydrolysis of yellow poplar wood meal. Ethanol fermentability in deacetylated yellow poplar hydrolysate (DYPH) by Pichia stipitis was also investigated. The alkali-pretreatment conditions were evaluated in terms of temperature, reaction time, and alkalinity. 94% of the acetyl group in xylan of the yellow poplar hemicellulose fraction was extracted using 0.5% sodium hydroxide solution at 60 degrees C for 60 min. The cell growth and ethanol production of P. stipitis was strongly affected by acetic acid, either in synthetic medium with 7.1g/l of acetic acid added or in yellow poplar hydrolysate (YPH) containing 7.1g/l of acetic acid. On the other hand, ethanol production in DYPH was slightly higher than that of the control although cell growth decreased by 34%. In the case of DYPH, the ethanol yield, volumetric ethanol productivity, and theoretical yield percentage was 0.48 g/g, 0.40 g/lh, and 93.2%, respectively. Thus, the alkaline-pretreatment method greatly enhanced the ethanol fermentability of yellow poplar hydrolysate.

Cystadenoma of the seminal vesicle.

Primary tumors of the seminal vesicle are quite rare with most reported cases being carcinomas. However, benign tumors of the seminal vesicle are extremely rare. We report a case of a cystadenomas of the seminal vesicles in a 46-year-old asymptomatic man, which was detected incidentally by computed tomography.

Characterization of Pb2+ biosorption from aqueous solution by Rhodotorula glutinis.

The yeast Rhodotorula glutinis was examined for its ability to remove Pb(2+) from aqueous solution. Within 10 min of contact, Pb(2+) sorption reached nearly 80% of the total Pb(2+) sorption. The optimum initial pH value for removal of Pb(2+ )was 4.5-5.0. The percentage sorption increased steeply with the biomass concentration up to 2 g/l and thereafter remained more or less constant. Temperature in the range 15-45 degrees C did not show any significant difference in Pb(2+ )sorption by R. glutinis. The light metal ions such as Na(+), K(+), Ca(2+), and Mg(2+) did not significantly interfere with the binding. The Langmuir sorption model provided a good fit throughout the concentration range. The maximum Pb(2+ )sorption capacity q(max) and Langmuir constant b were 73.5 mg/g of biomass and 0.02 l/mg, respectively. The mechanism of Pb(2+) removal by R. glutinis involved biosorption by direct biosorptive interaction with the biomass through ion exchange and precipitation by phosphate released from the biomass.