Comparison of lipase-catalysed esterification in supercritical carbon dioxide and in n-hexane

Summary Oleic acid esterification by ethanol has been performed by an immobilized lipase fromMucor miehei in supercritical carbon dioxide and in n-hexane as solvents. In both media, determination of apparent kinetic constants has been achieved and influence of water content has been shown to be different due to various rates of water solubilities. Stability of the lipase has been proved to be correct and similar in both solvents. Inhibition by ethanol excess has been found but is greater in n-hexane. That can explain the higher initial velocities obtained in supercritical carbon dioxide for the highest ethanol concentrations.     

Cloning, mapping and gene product identification of rhaT from Escherichia coli K12

Rhamnose utilization requires the function of a specific rhamnose transport system. Rhamnose transport mutants have been isolated and characterized. The structural gene, rhaT, encoding the rhamnose permease has been cloned from Escherichia coli. rhaT has been mapped in the rha locus (87.7 min) by analysis of cotransduction with glpK and other rha markers. The precise location of the gene has been determined by complementation analysis of rhamnose transport mutants transformed with recombinant plasmids containing different fragments of the cloned region. Gene order (counterclockwise) is established as glpK . . . rhaT-rhaR-rhaS-rhaB-rhaA-rhaD. The gene product has been identified by expression of rhaT in a T7 RNA polymerase/promoter system. This 23 kDa protein has been assigned to the rhaT product and has been shown to be located in the cell membrane.     

Productivity improvement in xanthan gum fermentation using multiple substrate optimization

A novel and more comprehensive formulation of the optimal control problem that reflects the operational requirements of a typical industrial fermentation has been proposed in this work. This formulation has been applied to a fed-batch bioreactor with three control variables, i.e., feed rates of carbon source, nitrogen source, and an oxygen source, to result in a 148.7% increase in product formation. Xanthan gum production using Xanthomonas campestris has been used as the model system for this optimization study, and the liquid-phase oxygen supply strategy has been used to supply oxygen to the fermentation. The formulated optimization problem has several constraints associated with it due to the nature of the system. A robust stochastic technique, differential evolution, has been used to solve this challenging optimization problem. The infinite dimensional optimization problem has been approximated to a finite dimensional one by control vector parametrization. The state constraints that are path constraints have been addressed by using penalty functions and by integrating them over the total duration to ensure a feasible solution. End point constraints on final working volume of the reactor and on the final residual concentrations of carbon and nitrogen sources have been included in the problem formulation. Further, the toxicity of the oxygen source, H(2)O(2), has been addressed by imposing a constraint on its maximum usable concentration. In addition, the initial volume of the bioreactor contents and feed concentrations have been handled as decision variables, which has enabled a well-grounded choice for their values from the optimization procedure; adhoc values are normally used in the industry. All results obtained by simulation have been validated experimentally with good agreements between experimental and simulated values.     

Light birefringence and molecular orientation in erythrocyte membranes and surface layers

The light birefringence in the surface layer of erythrocytes has been determined. A mathematical expression for the phase difference caused by birefringence in the spherical layer has been derived. Thickness of the anisotropic layer of erythrocytes has been evaluated. Birefringence in surface layers of erythrocytes has been shown to be caused not only by the membrane, but also by close to it structured proteins.     

The effect of oligomycin on rat liver mitochondria respiring in state 4

It has been found that oligomycin inhibits up to at least 50% state-4 mitochondrial respiration. A time dependence of oligomycin inhibition has been shown. A titration curve for state-4 respiration of sigmoidal profile has been presented. The possibility of misreading this oligomycin effect, so far never reported, has been excluded by evaluating the quality of mitochondrial preparations used in respect to their morphological, functional and electrochemical properties. The conclusion has therefore been put forward that the most part of respiration in steady-state-4 is driven by ATP synthesis.     

Chlorogenic acid in a Nicotiana plumbaginifolia cell suspension

A phenylpropanoid compound has been characterized in a Nicotiana plumbaginifolia cell suspension. This compound has been isolated and purified by semi-preparative reverse phase-high performance liquid chromatography. Its structure has been identified by NMR spectroscopy as 5-O-caffeoylquinic acid, which is chlorogenic acid (CA). The influence of culture conditions on the accumulation of this metabolite by N. plumbaginifolia cell suspensions has been studied. Darkness strongly inhibits the CA accumulation. Moreover, it has been shown that feeding experiments with caffeic acid had a deleterious effect upon the CA content. This one was not influenced by a supplementation with quinic acid.     

Gramicidin S-synthetase. Preparation of the multienzymic complex with a high specific activity.

A new purification procedure for the multienzyme of gramicidin S-synthetase has been developed. In vitro proteolysis with partial inactivation is suppressed by protease inhibitors EDTA, phenylmethylsulfonylfluoride, and fast preparation methods during initial separation steps. Activity has only been assayed by the total reaction of gramicidin S-synthetase, not by partial reactions of amino acid activation. The assay has been improved by evaluation of inhibitory concentrations of buffers, salts, and the product gramicidin S. It has been demonstrated that the rate of peptide synthesis in extracts containing both enzymes of gramicidin S-synthetase depends on protein concentration in a second order function. The multienzyme or heavy enzyme has been purified about 1400-fold to a specific activity of 24 nM/min per mg of protein, and the relation of this activity to the calculated in vivo activity is discussed.     

人类活动影响下新疆生态环境的一些变化

人类活动引起的新疆生态环境变化主要有以下几方面:1.河流流程缩短,河水矿化度和泥沙含量增加;2.湖泊水位下降,湖面缩小或变干,湖水矿化度升高;3.森林破坏,面积缩小;4.草场的产草量降低,牧草质量下降; 5.土壤次生盐渍化、沼泽化和肥力下降;6.土地沙漠化不断扩大。     

Fertility factors in Pseudomonas putida: selection and properties of high-frequency transfer and chromosome donors.

The octane plasmid (OCT) in Pseudomonas putida strains has been shown to be transferred at low frequency. However, bacteria which had newly received this plasmid showed a transient increase in donor ability. Using Octane+ P. putida as the donor, the transfer of most chromosomal markers was shown to be independent of OCT transfer, whereas the mobilization of the octanoate catabolism genes (octanoic and acetate) was dependent on OCT plasmid transfer. The presence of a fertility factor termed FPo has been postulated to explain these results. Strains carrying only this fertility factor have been obtained from strains carrying both OCT and FPo plasmids. Strains in which the OCT plasmid was transferred at high frequencies have also been isolated, and chromosome mobilization by OCT and FPo has been compared. A different gradient of transmission by OCT and FPo has been observed. It has also been shown that chromosome transfer by OCT was dependent on the bacterial recombination system, whereas the chromosome transfer by FPo was unaffected by the presence of a rec mutation in the donor strain.     

Purification and substrate specificity of BcmI restriction endonuclease

A strain producing the new specific restriction endonuclease BcmI has been found in the Bacillus generum. The enzyme has been purified by chromatography on the blue sepharose, phosphocellulose PII, heparin sepharose. The analogous purification has been obtained when the blue sepharose has been substituted for the orange sepharose, the home produced sorbent. The BcmI enzyme has been shown by the substrate specificity definition to be an isoschizomer of the restriction endonuclease ClaI.     

Construction of recombinant plasmids encoding the biosynthesis of the beta-subunit of cholera toxin

The structure of the cholera toxin operon and the location of A and B toxin subunits have been studied by the Southern blot hybridization on filters. The gene coding for the synthesis of the cholera toxin B-subunit has been cloned in the vector plasmid pBR322. The structural gene of A-subunit has been partially deleted by the restriction endonuclease Bal31 digestion. The size of the 250 b. p. deletion has been defined by electron microscopy. The production of the cholera toxin B-subunit in Escherichia coli K12 cells has been studied.     

Determination of amylase activity in bovine serum and its association with the Ami and Amll loci

Amylase activity has been estimated in the sera of 213 cattle by an amyloclastic method with potato amylose as the substrate with the addition of CaCl₂. The method has been developed from that of Street and Close to overcome the problems of the high colour of bovine serum which would otherwise interfere with the assay. A relationship has been found between the serum amylase genotype detected on starch-gel and the total enzyme activity. The enzymes AmI-B and AmII-B in homozygous combinations have been found to have the highest activities. Age of animal has no significant effect on the total enzyme activity. Activity has been investigated in sera from three separate cattle populations and it has been found to be a highly heritable character. Heritability estimates of 49 % and 39 % by sib analysis and 75 % by the twin method have been obtained.     

一种植物生长激素-N'-(5-硝基-2-羟基苯亚甲基)-3,5-二羟基苯甲酰肼的合成、晶体结构和抗菌活性

目的:合成并表征一种属于植物生长激素的席夫碱类化合物.方法:由5-硝基水杨醛和3,5-二羟基苯甲酰肼在甲醇溶液中反应得到N′-(5-硝基-2-羟基苯亚甲基)-3,5-二羟基苯甲酰肼.结果:用单晶X-射线衍射法测得其结构并表征,同时通过抗菌实验证明其在一定浓度下具有明显的抗菌活性.结论:这种席夫碱类化合物在某种程度上具有明显的抗菌活性.     

Retinoid-binding proteins are phosphorylated in vitro by soluble Ca+2- and phosphatidylserine-dependent protein kinase from mouse brain

A direct radioimmunoassay of atrial natriuretic factor (ANF) has been developed. The method uses a synthetic 26 amino-acid fragment (8-33 ANF) of the native peptide. Antibodies have been prepared in rabbits immunized with the peptide coupled to thyroglobulin. The radiolabelled tracer prepared by iodination according to the Chloramine-T method has been purified by HPLC followed by affinity chromatography on Sepharose-4B anti-ANF. Dextran-coated charcoal has been used for separation of free from antibody bound radioactivity. Higher ANF content has been found in the right rat atrium than in the left. These results have been confirmed by bioassay.     

Transmembrane potential of liver mitochondria from hexachlorobenzene-and iron-treated rats

The respiratory parameters and the membrane potential of liver mitochondria from rats treated with either hexachlorobenzene, iron or hexachlorobenzene plus iron, to induce experimental porphyria, have been studied. Partial uncoupling of oxidative phosphorylation has been observed in mitochondria from hexachlorobenzene- and hexachlorobenzene plus iron-treated rats. Direct evidence has been presented that this uncoupling is due to the action of pentachlorophenol endogenously formed by metabolism of hexachlorobenzene. No irreversible damage of mitochondria membrane has been revealed under both these conditions. Normal oxidative phosphorylation has been found in mitochondria from rats treated with iron alone. In contrast, they presented an anomalous membrane potential, fully restored by oligomycin. A possible involvement of lipid peroxidation process, induced by iron, in causing these abnormalities has been suggested.     

pH-sensitive hydrogel based on a novel photocross-linkable copolymer

A pH sensitive hydrogel has been prepared by a UV irradiation technique. Starting polymer was the PHM (poly hydroxyethylaspartamide methacrylated) obtained from polyaspartamide (PHEA) partially derivatized with methacrylic anhydride (MA). This new copolymer has been further derivatized with succinic anhydride (SA) to obtain PHM-SA that has been cross-linked by UV irradiation to form a pH sensitive hydrogel. The network, recovered after washing as a powder, has been been characterized by FT-IR spectrophotometry and particle size distribution analysis. Moreover, to have information about water affinity of the prepared sample, swelling measurements have been carried out in aqueous media mimicking biological fluids. The possibility to employ the prepared hydrogel as a pH-sensitive drug delivery system (DDS) has been investigated. In particular, ibuprofen ((S)(+)4-isobutyl-alpha-methylphenyl-acetic acid), chosen as a model drug, has been entrapped into the PHM-SA hydrogel, and in vitro release studies have showed that its release rate depends on different swelling of the network as a function of the environmental pH.     

Degradation of polysaccharides and lignin in wood ozonation

Ozonation has been considered as a method for the pretreatment of plant biomass to obtain cellulose and monosaccharides. Ozone consumption by aspen wood with various moisture contents has been investigated. We have considered the gradual transformation of the substrate: wood to ozonated wood to cellulose-containing product (CP) to holocelluloze (HC) and to cellulose. Yields of ozonated wood (OW), the (CP), water-soluble ozonation products, HC, and cellulose have been determined. The lignin content in the CP has been estimated. Both HC and cellulose samples have been studied by IR spectroscopy. The degree of polymerization and molecular mass distribution of cellulose obtained from ozonated wood have been determined. It has been shown that wood destruction by ozone is accompanied by degradation of lignin, hemicelluloses, and cellulose.It has been found that physicochemical properties of cellulose obtained from ozonated wood can be regulated by the variation of the initial moisture content in the substrate. Both molecular ozone and radical species, which are generated in the course of ozone reactions with water present in the substrate structure, participate in wood destruction.     

5'-methylthioadenosine phosphorylase from the human prostate. 1. Purification and partial characterization]

5'-Methylthioadenosine phosphorylase has been purified approximately 340-fold in 20% yield from human prostate: the use of affinity chromatography by Sepharose-Hg has been found particularly advantageous. The enzyme has been partially characterized and an apparent Km of 2.5 x 10(-5) M has been calculated for 5'-methylthioadenosine. The reaction is activated by thiols and shows an absolute requirement for phosphate ions.     

Thermodynamic and kinetic study of thermal denaturation of Kunitz soybean trypsin inhibitor by differential scanning microcalorimetry

The thermal denaturation of soybean trypsin inhibitor (Kunitz inhibitor) has been studied in pH-region from 2.0 to 11.0 by differential scanning microcalorimetry. The thermodynamic characteristics have been determined. It has been established that the denaturation transition of protein may be described by a two-state model. It has been shown, that two side hydrogen bonds between carboxylate-ion and tyrosyl and carboxylate-ion and lysyl take part in the stabilization of the inhibitor's native structure. The activation of denaturation is accompanied by cleavage of one side hydrogen bond.