Analysis of methanol–ethanol mixtures from falsified beverages using a dual biosensors amperometric system based on alcohol dehydrogenase and alcohol oxidase

A portable bienzymatic analytical system was developed for the chronoamperometric analysis of methanol–ethanol mixtures. The system consists of two biosensors, one based on alcohol dehydrogenase (ADH) that responds only to the ethanol and the second one based on alcohol oxidase (AOX) that responds to both methanol and ethanol. The transducers were screen-printed electrodes (SPEs) modified with mediators: Meldola blue for ADH and Co-phthalocyanine for AOX. The calibration graph of the ADH biosensor is linear between 0.3 and 8 mmol/L ethanol. The AOX biosensor is able to quantify both analytes in mixtures that contain methanol between 3 and 70 mmol/L and ethanol ranging from 15 to 110 mmol/L. Interferences due to non-specific oxidations from common oxidizable compounds like gallic acid and ascorbic acid were smaller in the case of transducer based on Meldola blue. The analytical system was successfully tested on real samples: non-alcoholised beer (NAB) spiked with ethanol or methanol and a falsified rose wine (FRW).     

Structural studies of enzyme-based microfluidic biofuel cells

An enzyme-based glucose/O₂ biofuel cell was constructed within a microfluidic channel to study the influence of electrode configuration and fluidic channel height on cell performance. The cell was composed of a bilirubin oxidase (BOD)-adsorbed O₂ cathode and a glucose anode prepared by co-immobilization of glucose dehydrogenase (GDH), diaphorase (Dp) and VK₃-pendant poly-l-lysine. The consumption of O₂ at the upstream cathode protected the downstream anode from interfering O₂ molecules, and consequently improved the cell performance (maximum cell current) ca. 10% for the present cell. The cell performance was also affected by the channel height. The output current and power of a 0.1 mm-height cell was significantly less than those of a 1 mm-height cell because of the depletion of O₂, as determined by the shape of the E–I curve at the cathode. On the other hand, the volume density of current and power was several times higher for the narrower cell.     

Phytoremediation of petroleum polluted soil

An experimental study of the rhizosphere effect on phytoremediation of petroleum polluted soil was carried out with three species of grasses, namely Pannicum, Eleusine indica （L.） Gaerth, and Tall Fescue. After a period of 150 days, this pot experiment showed that the rhizosphere of these three species accelerated the degradation of petroleum hydrocarbons to different extents. The results showed that the number of microorganisms in the rhizosphere increased by three orders of magnitude. The induction of the plant rhizosphere and the coercion influence of petroleum changed the species and activity of microorganisms. The degradation of petroleum hydrocarbons in the rhizosphere was 3-4 times that in unplanted soil. The dehydrogenase activity in the rhizosphere was 1.61-2.20 times that in unplanted soil, but the catalase activity was 0.90-0.93 times that in unplanted soil, and soil moisture content increased by 5% compared with the unplanted soil.     

Interaction of CO dehydrogenase with the cytoplasmic membrane monitored by fluorescence correlation spectroscopy

We have investigated the interaction of carbon monoxide dehydrogenase (CODH), an enzyme that catalyses the oxidation of CO in the aerobic eubacterium Oligotropha carboxidovorans, with the cytoplasmic membrane by using fluorescence correlation spectroscopy (FCS). Our results reveal that in vitro this interaction of CODH is specific for cytoplasmic membranes from CO-grown bacteria.     

Inhibition of Human DHODH by 4‐Hydroxycoumarins,Fenamic Acids,and N‐(Alkylcarbonyl)anthranilic Acids Identified by Structure‐Guided Fragment Selection

A strategy that combines virtual screening and structure‐guided selection of fragments was used to identify three unexplored classes of human DHODH inhibitor compounds: 4‐hydroxycoumarins, fenamic acids, and N‐(alkylcarbonyl)anthranilic acids. Structure‐guided selection of fragments targeting the inner subsite of the DHODH ubiquinone binding site made these findings possible with screening of fewer than 300 fragments in a DHODH assay. Fragments from the three inhibitor classes identified were subsequently chemically expanded to target an additional subsite of hydrophobic character. All three classes were found to exhibit distinct structure–activity relationships upon expansion. The novel N‐(alkylcarbonyl)anthranilic acid class shows the most promising potency against human DHODH, with IC₅₀ values in the low nanomolar range. The structure of human DHODH in complex with an inhibitor of this class is presented.     

Antioxidative activities of bran extracts from twenty one pigmented rice cultivars

Ethanol–water (70:30, v/v) extracts from the bran of rice seeds from twenty one pigmented and one nonpigmented rice cultivars were evaluated for antioxidative activities using the following tests: inhibition of peroxidation of linoleic acid; inhibition of peroxidation of rabbit lipid erythrocyte membranes; reduction of potassium ferricyanide, and scavenging of superoxide anions and hydroxyl radicals. With some exceptions, extracts from the pigmented rice seeds had higher antioxidative activity than did the nonpigmented variety. The following pigmented cultivars had the highest antioxidative activities in all tests: Jumlalocal-1, Parnkhari 203, DZ78, LK1-3-6-12-1-1, and Elwee. A significant correlation was also noted between reducing power, inhibition of erythrocyte ghost membrane peroxidation, and superoxide anion and hydroxyl radical scavenging. The results suggest that: (a) ferricyanide reducing power might be a useful and simple index for large-scale evaluation of antioxidative potencies of natural products present in rice; (b) pigmented rice varieties with high antioxidative activities provide a source of antioxidants and a genetic resource to develop new health-promoting rice cultivars.     

Effect of gene disruption of succinate dehydrogenase on succinate production in a sake yeast strain

Succinate dehydrogenase (SDH) of Saccharomyces cerevisiae consists of four subunits encoded by the SDH1, SDH2, SDH3, and SDH4 genes. We determined the effect of SDH deficiency on the productivity of organic acids in a sake yeast strain Kyokai no. 9. The SDH activity of single disruptants was retained at 30-90% of that of the wild-type strain, but the activity disappeared in double disruptants of the SDH1 and SDH2 or SDH1b (the SDH1 homologue) genes. Two double disruptants showed no growth on a medium containing glycerol as the sole carbon source, while the single disruptants could utilize glycerol. These results indicate that double disruption of the SDH1 and SDH2 or SDH1b genes is required for complete loss of SDH activity and that the SDH1b gene compensates for the function of the SDH1 gene. The sdh1 sdh1b disruptant showed a marked increase in succinate productivity of up to 1.9-fold along with a decrease in malate productivity relative to the wild-type strains under shaking conditions. Under both static and sake brewing conditions, the productivity of these organic acids in the disruptants was virtually unchanged from that in the wild-type strain. Furthermore, SDH activity was undetectable in the wild-type and the disrupted strains under static conditions. These results suggest that SDH activity contributes to succinate production under shaking conditions, but not under static and sake brewing conditions.     

生物法处理有机废水的实验研究

本采用序列间歇式反应器（SBR）研究了生物法处理制药有机废水的可行性。实验结果表明，在进水COD为523－2149mg/L，曝气16h时，COD去除率为75－90％，出水COD可达到国家行业废水排放标准。废水中有机污染物通过活性污泥微生物作用被降解，COD浓度降低，基质性脱氢酶活性（DHAs)水平也降低。总脱氢酶活性与活性细菌数目（ABN）的对数显相关。     

乳酸脱氢酶高产菌株的选育及条件研究

以植物乳酸短杆菌RS2-2(Lactobacillus plantarum)为出发菌株,经紫外线、亚硝基胍单独处理和复合处理,获得1株乳酸脱氢酶高产菌株U-N-B14,通过优化实验对该菌株生产乳酸脱氢酶的营养要求和发酵条件进行了研究,优化后的发酵培养基组成为(g/L):酵母膏10,麦芽糖12,乙酸铵4,K2HPO44,NaCl 8,吐温80 2mL/L。发酵条件是:培养温度30℃,pH值6.5,培养时间24 h,接种量8%。在此条件下,1.5 m3罐中试生产的乳酸脱氢酶产量可达1 497 U/g。