Effect of oxygen abstraction on the peroxidase–luminol–perborate system: Relevance to the HRP enhanced chemiluminescence mechanism

Abstraction of oxygen from the HRP enhanced chemiluminescence system has no significant effect on the chemiluminescence generated. It is, therefore, proposed that in the peroxidase-luminol-perborate system at pH 7.3, chemiluminescence is generated by a direct reaction of diazaquinones with hydrogen peroxide and not, as generally assumed, from the reaction of luminol radicals with the molecular oxygen.     

亚洲薄荷的两个化学型

亚洲薄荷的两个化学型桂新,周荣汉（安徽中医学院中药系合肥２３００３８）（中国药科大学植物化学分类研究室南京２１００３８）ＴｗｏｃｈｅｍｏｔｙｐｅｓｏｆＭｅｎｔｈａａｓｉａｆｉｃａＢｏｒｉｓｓ￥ＣｈｏｕＧｕｉ－Ｘｉｎ（ＡｎｈｕｉＣｏｌｌｅｇｅｏｆＴｒａ...     

Esterification of 2-methylalkanoic acids Catalysed by Lipase from Candida rugosa: Enantioselectivity as a Function of water Activity and Alcohol Chain Length

Esterifications catalysed by immobilised lipase from Candida rugosa (CRL) in cyclohexane at constant water activity (a_w = 0.76) were studied using 2-methyl substituted octa-, nona- or decanoic acids and n-alcohols of varying chain length as substrates. The importance of controlling the water activity and choosing the right alcohol for obtaining maximum enantioselectivity is demonstrated. The immobilised lipase was easily recovered without loss of activity and enantioselectivity.     

The placement, recovery, and loss of scatter hoards by eastern chipmunks,Tamias striatus

Although eastern chipmunks typically larder hoard food in theirburrows, some scatter hoarding occurs, especially by newly emergedjuveniles and by females with young. To examine patterns ofscatter hoard placement and their adaptive significance, weused standard food provisioning tests followed by continuousmonitoring to determine the longevity and fate of scatter hoards.Longevity of scatter hoards was low (median 74 min) and pilferagewas high (46%). Chipmunks placed scatter hoards away from thepatch, closer to their burrow and within the defended area ofprimary use. Scatter hoard placement was affected by the numberof competitors at the patch. However, juveniles and femaleswith young differed slightly in their responses. competitorsthat approached scatteihoards were sometimes chased away bythe scatter hoard's owner, and the scatter hoard was moved toanother location by the owner. Females with young recoveredtheir largest scatter hoards first, but there was little additionalpattern in scatter hoard recovery. An experiment using simulatedscatter hoards showed that scatter hoards farther from a patchof food lasted longer and that disturbing and marking scatterhoards reduced their longevity. These observations indicate:(1) that scatter hoard placement is more related to avoidingpilferage of completed caches than to rapidly sequestering fooditems from an ephemeral patch and (2) that optimal cache distributionmay be affected by the cache-defense ability of individuals,by the relationship between competitor density and cache securitynear the patch, and by the territorial behavior of neighboringconspecifics.     

Physico-chemical characteristics of a permanent Spanish hypersaline lake: La Salada de Chiprana (NE Spain)

La Salada de Chiprana Lake, located in the Ebro River basin, northeastern Spain, is the only permanent and deep water hypersaline ecosystem in all of western Europe. With a total surface of 31 ha and a maximum depth of 5.6 m, it has several basins bounded by elongated sandstone-bodies or ribbons which are paleochannels of Miocene age. Its salinity varied from 30 to 73 g 1^–1 during the 1989 hydrological cycle and the most abundant ions were magnesium and sulphate. Depth-time distributions of major physico-chemical variables demonstrated that the lake was stratified in two distinctive layers during most of the year. The chemocline disappeared only in October, with the complete overturn of the water column. In the deep water, three conditions occurred which allowed development of green sulphur bacteria populations: (1) oxygen depletion, (2) presence of hydrogen sulphide and (3) presence of light. Benthic microbial mats covered the sediments of shallow shores of moderate slope.     

Molecular integrity of monoclonal antibodies produced by hybridoma cells in batch culture and in continuous-flow culture with integrated product recovery

The molecular integrity of monoclonal antibodies (MCAB) produced by murine hybridoma cell line TB/C3 was studied in batch and continuous-flow cultures. In batch culture, one band of MCAB was detected initially by Western blotting of sodium dodecyl sulfate (SDS)-polyacrylamide gels run under unreduced conditions, but heterogenous MCAB bands appeared as the culture aged. The latter were due to the degradation of MCAB by proteases active at the neutral pH of the culture. The deleterious effect of proteases was minimized in the continuous-flow cultures which were integrated for product recovery. The MCAB of high quality was purified over 26 days from a culture grown at a dilution rate of 0.025 h(-1) (experiment 1). However, at a lower dilution rate of 0.015 h(-1) (experiment 2), the integrity of MCAB was compromised after the initial 13 days of culture. This was shown to be due to the variation in the carbohydrate content of MCAB produced, as judged by the increased sialylation of heavy chains and the varied reactivity of MCAB with lectins (Maackia amurensis agglutinin, Galanthus nivalis agglutinin, and Datura stramonium agglutinin) as the age of the culture increased. The concentration of the purified MCAB samples by enzyme-linked immunosorbent assay (ELISA) (used normally) was usually higher than that estimated by absorbance at 280 nm. Best correlation between the two methods (ELISA-280 nm ratio of 1.02-1.25) was obtained with experiment 1 samples. This ratio increased in experiment 2 and batch culture samples as the heterogeneity of MCAB produced increased, being 1.03-2.94 and 2.53-4.62, respectively. Therefore, ELISA overestimated MCAB concentration when the molecular integrity of the latter was compromised. The ELISA-A(280) nm ratio might hence provide a useful indicator for assessing the quality of MCAB produced. Comparison of SDS-polyacrylamide gels stained with Coomassie Brilliant Blue R and silver showed that the former correlated better with the MCAB activity stain, whereas the silver stained both the protein- and carbohydrate-rich components. Comparison of the patterns produced with these two stains might therefore offer another parameter to monitor the overall integrity of MCAB produced. Finally, the data presented have important implications on the validity of using long-term and intensive cultures for generating MCAB because such cultures would be subjected to the additive effects reported for batch and continuous modes of growth. (c) 1993 John Wiley & Sons, Inc.     

Emulsion liquid membrane extraction of lactic acid from aqueous solutions and fermentation broth

Studies on the batch extraction of lactic acid using an emulsion liquid membrane system are reported. The membrane phase consists of the tertiary amine carrier Alamine 336 and the surfactant Span 80 dissolved in n-heptane/paraffin and aqueous solutions of sodium carbonate in the internal phase. The effects of internal phase reagent, extraction temperature, and initial external phase pH on the extraction efficiency and the emulsion swelling are examined. A statistical factorial experiment on extraction from clarified lactic acid fermentation broth was carried out to obtain knowledge of the performance of the extraction system from a broth. The extraction efficiency from the fermentation broth is found to be lower as compared to aqueous solutions of pure lactic acid. The effect of pH and the presence of other ionic species on selectivity are discussed. (c) 1993 John Wiley & Sons, Inc.     

Development of a recovery and recycle process for a pseudomonas lipase used for large-scale enzymatic synthesis

Enzymes are potential catalysts for a wide range of large-scale chemical synthesis steps, particularly when the creation of a specific chiral center is desired. The efficient recycling of the enzyme catalyst and the removal of carryover impurities were crucial factors in the improvement of a stereoselective ester hydrolysis step used in the synthesis of a selective leukotriene antagonist. In this enzymatic reaction step, the substrate and product were both largely insoluble, while the enzyme was soluble in the aqueous reaction mixture. Microfiltration and ultrafiltration of the slurry reaction mother liquor indicated near 100% enzyme protein recovery, while activity recovery was about 70% to 80%. These activity losses might be accounted for by enzyme degradation (1 to 2 mg/L . h) during the 40-hour reaction period. Dissolved impurities, principally a diacid byproduct, in the enzyme recycling stream were reduced 60% to 70% by either lowering the solution pH to 4.0 or raising the solution ionic strength to 1 M. (c) 1993 John Wiley & Sons, Inc.     

Continuous measurement of microbial heat production in laboratory fermentors

The possibility of continuously measuring the heat produced by microorganisms in an ordinary laboratory fermentor was studies. An inventory of the heat flows influencing the temperature of the culture was made. The magnitude and standard deviation in these heat flows were studied from theoretical and practical viewpoints. Calibration procedures were tested, and a model describing the heat flows in steady state and during dynamic conditions was made. Microbial heat production could be calculated accurately with the help of this model, appropriate temperature measurements, and equipment properties measured during the calibration procedures. It was found that the measurement of heat production could be done with an accuracy similar to that in the O(2) uptake measurement. (c) 1993 John Wiley & Sons, Inc.     

Kinetics of competitive inhibition and cometabolism in the biodegradation of benzene, toluene, and p-xylene by two Pseudomonas isolates

Two Pseudomonas species (designated strains B1 and X1) were isolated from an aerobic pilot-scale fluidized bed reactor treating groundwater containing benzene, toluene, and p-xylene (BTX). Strain B1 grew with benzene and toluene as the sole sources of carbon and energy, and it cometabolized p-xylene in the presence of toluene. Strain X1 grew on toluene and p-xylene, but not benzene. In single substrate experiments, the appearance of biomass lagged the consumption of growth substrates, suggesting that substrate uptake may not be growth-rate limiting for these substrates. Batch tests using paired substrates (BT, TX, or BX) revealed competitive inhibition and cometabolic degradation patterns. Competitive inhibition was modeled by adding a competitive inhibition term to the Monod expression. Cometabolic transformation of nongrowth substrate (p-xylene) by strain B1 was quantified by coupling xylene transformation to consumption of growth substrate (toluene) during growth and to loss of biomass during the decay phase. Coupling was achieved by defining two transformation capacity terms for the cometabolizing culture: one that relates consumption of growth substrate to the consumption of nongrowth substrate, and second that relates consumption of biomass to the consumption of nongrowth substrate. Cometabolism increased decay rates, and the observed yield for strain B1 decreased in the presence of p-xylene. (c) 1993 Wiley & Sons, Inc.