Haloperoxidases: Enzymatic Synthesis of alpha,beta-Halohydrins from Gaseous Alkenes

The enzymatic synthesis of alpha,beta-halohydrins from gaseous alkenes is described. The enzymatic reaction required an alkene, a halide ion, dilute hydrogen peroxide, and a haloperoxidase enzyme. A wide range of gaseous alkenes were suitable for this reaction, including those containing isolated, conjugated, and cumulative carbon-carbon double bonds. Chlorohydrins, bromohydrins, and iodohydrins could be formed. The combining of this enzymatic synthesis with a previously described enzymatic synthesis of epoxides from alpha,beta-halohydrins provides an alternate pathway, other than the well-known enzymatic direct epoxidation pathway, from alkene to an epoxide.     

Haloperoxidases: Enzymatic Synthesis of α,β-Halohydrins from Gaseous Alkenes

The enzymatic synthesis of α,β-halohydrins from gaseous alkenes is described. The enzymatic reaction required an alkene, a halide ion, dilute hydrogen peroxide, and a haloperoxidase enzyme. A wide range of gaseous alkenes were suitable for this reaction, including those containing isolated, conjugated, and cumulative carbon-carbon double bonds. Chlorohydrins, bromohydrins, and iodohydrins could be formed. The combining of this enzymatic synthesis with a previously described enzymatic synthesis of epoxides from α,β-halohydrins provides an alternate pathway, other than the well-known enzymatic direct epoxidation pathway, from alkene to an epoxide.     

有机介质中酶催化活性和选择性的调控

有机溶剂中酶的结构与功能与在水中有很大的不同,通过调整控制策略可系统地改善酶针对目标反应的活性和选择性,重点阐述了溶剂对酶催化反应的活性和选择性的影响,介绍了酶催化选择性的热力学预测模型。,     

Comparing impacts of physicochemical properties and hydrolytic inhibitors on enzymatic hydrolysis of sugarcane bagasse

An autohydrolysis pretreatment with different conditions was applied to sugarcane bagasse to compare the impacts of the physicochemical properties and hydrolytic inhibitors on its enzymatic hydrolysis. The results indicate that the autohydrolysis conditions significantly affected the physicochemical properties and inhibitors, which further affected the enzymatic hydrolysis. The inhibitor amount, pore size, and crystallinity degree increased with increasing autohydrolysis severity. Furthermore, the enzymatic hydrolysis was enhanced with increasing severity owing to the removal of hemicellulose and lignin. The physicochemical obstruction impeded the enzymatic hydrolysis more than the inhibitors. The multivariate correlated component regression analysis enabled an evaluation of the correlations between the physicochemical properties (and inhibitors) and enzymatic hydrolysis for the first time. According to the results, an autohydrolysis with a severity of 4.01 is an ideal pretreatment for sugarcane bagasse for sugar production.

     

The use of an enzymatic kit to measure plasma creatinine in the mouse and three other species

Plasma creatinine concentrations were determined in mice, rats, dogs and humans using a kinetic alkaline picrate method and an enzymatic method with a centrifugal analyser. Lower creatinine values were obtained for all four species using the enzymatic method, but the differences between methods were greater for mice and rats. Removal of creatininase from the enzymatic method reagents gave proportionally higher values for non-creatinine chromogens in mice and rats. A single enzymatic method was not specific for creatinine when used for these species.     

Some notes on the theory of reaction rates: Enzymatic reactions

A generalized model of an enzymatic reaction in a biological system is used for the construction of a possible model of the “concentrating mechanism” after the introduction of a tracer, and for the formulation of an excretion rate of substances reabsorbed or actively secreted into the urine by the kidneys. In considering this generalized model of the enzymatic reaction, the term “tracer” is used in a limited sense, and it is concluded that a linear system is suitable for the study of the kinetics of the tracer—even in the case when the transport of the tracer between compartments is due to an enzymatic reaction. The rate coefficients of the linear differential systems, which are dependent on the state of the enzymatic system, are functions of the amount of the substrates, as well as of the amounts of activators and inhibitors of the enzymatic reaction and of the amount of the substances, which are responsible for the enzyme production.     

Enzymatic membrane reactors for biodegradation of recalcitrant compounds. Application to dye decolourisation

Membrane bioreactors are being increasingly used in enzymatic catalysed transformations. However, the application of enzymatic-based treatment systems in the environmental field is rather unusual. The aim of this paper is to overview the application of enzymatic membrane reactors to wastewater treatment, more specifically to dye decolourisation. Firstly, the basic aspects such as different configurations of enzymatic reactors, advantages and disadvantages associated to their utilisation are revised as well as the application of this technology to wastewater treatment. Secondly, dye decolourisation by white-rot fungi and their oxidative enzymes are discussed, presenting an overall view from for in vivo and in vitro systems. Finally, dye decolourisation by manganese peroxidase in an enzymatic membrane reactor in continuous operation is presented.     

生物酶法合成L-精氨酸衍生物的研究进展

L-精氨酸是一种碱性氨基酸,具有多样化的官能团,是合成多种有用化合物的前体,其衍生物广泛应用于医疗、食品和化妆品等领域。L-精氨酸衍生物的合成方法有化学法、发酵法和酶法。在当前绿色经济和可持续发展的背景下,对比各种生产方法,生物酶法合成L-精氨酸衍生物具有明显优势。因此本文重点介绍了L-精氨酸衍生化的典型产品和合成技术,并介绍了生物酶法合成L-精氨酸衍生物未来可能的发展方向。     

pH catalyzed pretreatment of corn bran for enhanced enzymatic arabinoxylan degradation

Corn bran is mainly made up of the pericarp of corn kernels and is a byproduct stream resulting from the wet milling step in corn starch processing. Through statistic modeling this study examined the optimization of pretreatment of corn bran for enzymatic hydrolysis. A low pH pretreatment (pH 2, 150 °C, 65 min) boosted the enzymatic release of xylose and glucose and maximized biomass solubilization. With more acidic pretreatment followed by enzymatic hydrolysis the total xylose release was maximized (at pH 1.3) reaching ～ 50% by weight of the original amount present in destarched corn bran, but the enzyme catalyzed xylose release was maximal after pretreatment at approx. pH 2. The total glucose release peaked after pretreatment of approx. pH 1.5 with an enzymatic release of approx. 68% by weight of the original amounts present in destarched corn bran. For arabinose the enzymatic release was negatively affected by the acidic pretreatment as labile arabinosyl-linkages were presumably hydrolysed directly during the pretreatment. A maximum of 60% arabinose release was achieved directly from the optimal (acidic) pretreatment. The total content of diferulic acids, supposedly involved in the cross-linking of the arabinoxylan polymers, decreased by both alkaline and acidic pretreatment pH, with the loss by alkaline pretreatments being highest. No direct correlation between the enzymatic release of xylose and the content of diferulic acids in the substrate could be verified. On the contrary the enzymatic release of xylose was significantly correlated to the total release of arabinose, indicating that the degree of arabinosyl-substitutions on the xylan backbone is an essential parameter for enzymatic hydrolysis of corn bran arabinoxylan.     

植物酶系统对UV-B辐射的响应机制

紫外辐射作为一种自然界存在的环境因子,影响着植物的生长发育。在植物从太阳辐射中获得光照和热量的同时,不可避免地要受到UV-B辐射的胁迫。植物在长期对环境的适应中,在体内形成了防御系统,以保护自身的安全,酶系统是其中之一。植物酶系统在实施防护的同时也受到了UV-B辐射的影响,信号分子对调节酶响应UV-B胁迫起了关键作用。本文重点论述了UV-B辐射增强下,植物酶系统的变化、植物酶系统的响应机制以及酶系统与信号物质的关系,并对该领域未来的研究方向提出了展望。     

Gamma-glutamyltranspeptidase stimulates receptor activator of nuclear factor-kappaB ligand expression independent of its enzymatic activity and serves as a pathological bone-resorbing factor

A novel bone-resorbing factor was cloned using an expression cloning technique, which involved a Xenopus oocyte expression system and an assay for osteoclast formation. A candidate clone was isolated from a BW5147 mouse T-lymphoma cell cDNA library. Sequencing analysis identified the factor as gamma-glutamyltranspeptidase (GGT), which is an enzyme involved in glutathione metabolism. The addition of purified GGT protein to mouse bone marrow culture effectively induced formation of osteoclasts. An antibody against GGT inhibited osteoclast formation but not the enzymatic activity. We also demonstrated that an inactive form of GGT, the enzymatic activity of which had been blocked by chemical modification with a specific inhibitor, acivicin, supported osteoclast formation. These results indicate that GGT acts on osteoclast formation independent of its own enzymatic activity. Furthermore, both native GGT and inactive GGT stimulated the expression of the receptor activator of nuclear factor-kappaB ligand (RANKL) mRNA and protein from bone marrow stromal cells. This report is the first demonstration of a novel biological activity of GGT protein in a manner independent of its enzymatic activity.     

Biocatalytic generation of Mn(III)-chelate as a chemical oxidant of different environmental contaminants

The objective of this study was to investigate the enzymatic generation of the Mn(3+) -malonate complex and its application to the process of oxidizing several organic compounds. The experimental set-up consisted of an enzymatic reactor coupled to an ultrafiltration membrane, providing continuous generation of Mn(3+) -malonate from a reaction medium containing versatile peroxidase (an enzyme produced by Bjerkandera adusta strain BOS55), H(2) O(2) , MnSO(4,) and malonate. The effluent of the enzymatic reactor was introduced into a batch-stirred reactor to oxidize three different classes of compounds: an azo dye (Orange II), three natural and synthetic estrogens, and a polycyclic aromatic hydrocarbon (anthracene). The enzymatic reactor provided the Mn(3+) complex under steady-state conditions, and this oxidative species was able to transform the three classes of xenobiotics considerably (90-99%) with negligible loss of activity.     

The effect of delignification of forest biomass on enzymatic hydrolysis

The effect of delignification methods on enzymatic hydrolysis of forest biomass was investigated using softwood and hardwood that were pretreated at an alkaline condition followed by sodium chlorite or ozone delignification. Both delignifications improved enzymatic hydrolysis especially for softwood, while pretreatment alone was found effective for hardwood. High enzymatic conversion was achieved by sodium chlorite delignification when the lignin content was reduced to 15%, which is corresponding to 0.30-0.35 g/g accessible pore volume, and further delignification showed a marginal effect. Sample crystallinity index increased with lignin removal, but it did not show a correlation with the overall carbohydrate conversion of enzymatic hydrolysis.     

Dynamic theory of enzymatic catalysis]

Formal kinetics of an enzymatic reaction is considered in terms of a dynamic scheme of catalysis. Conformation reconstructions of a liberated enzyme and e-s-complex are analysed upon the phase plane. An expression for the enzyme stationary activity depending on the rate of conformation reconstructions is obtained. The case of purely dynamic catalysis is analysed and possible initiation of enzymatic activity variation in time connected with initial synchronization of molecules is shown. This phenomenon is considered as a possible criterion of the dynamic character of enzymatic catalysis.     

Enzyme Kinetics for Complex System Enables Accurate Determination of Specificity Constants of Numerous Substrates in a Mixture by Proteomics Platform

Many important experiments in proteomics including protein digestion, enzyme substrate screening, enzymatic labeling, etc., involve the enzymatic reactions in a complex system where numerous substrates coexists with an enzyme. However, the enzyme kinetics in such a system remains unexplored and poorly understood. Herein, we derived and validated the kinetics equations for the enzymatic reactions in complex system. We developed an iteration approach to depict the enzymatic reactions in complex system. It was validated by 630 time-course points from 24 enzymatic reaction experiments and was demonstrated to be a powerful tool to simulate the reactions in the complex system. By applying this approach, we found that the ratio of substrate depletion is independent of other coexisted substrates under specific condition. This observation was then validated by experiments. Based on this striking observation, a simplified model was developed to determine the catalytic efficiencies of numerous competing substrates presented in the complex enzyme reaction system. When coupled with high-throughput quantitative proteomics technique, this simplified model enabled the accurate determination of catalytic efficiencies for 2369 peptide substrates of a protease by using only one enzymatic reaction experiment. Thus, this study provided, in the first time, a validated model for the large scale determination of specificity constants which could enable the enzyme substrate screening approach turned from a qualitative method of identifying substrates to a quantitative method of identifying and prioritizing substrates. Data are available via ProteomeXchange with identifier PXD004665.     

Mitochondrial enzymatic adaptation of skeletal muscle to endurance training.

Some mitochondrial enzymatic activities (succinate dehydrogenase, NADH cytochrome reductase, cytochrome oxidase) were studied in the gastrocnemius and soleus muscle of the rat. The modifications of the enzyme activity, induced by endurance training, were found to be functions of 1) daily work load and 2) total training time. The treatment with an effective dose of vasodilating substances (papaverine, nicergoline, dipyridamole, and bamethan) showed that 1) nicergoline, bamethan, and dipyridamole were differently able to shorten the time of appearance of the increase in the enzymatic activities; 2) however, long-term treatments with these drugs did not prove able to modify the plateau level of the enzymatic activity increase, for a given amount of endurance training; 3) the pharmacodynamic effect on enzymatic activities was in no way related to the vasodilating effect of these drugs, since the effect was not observed with papaverine. The transition from a given level of endurance training to a lower one led to a proportional decrease of the mitochondrial enzymatic activities, thus pointing out the relation between amount of training and enzymatic activity. The drugs studied were unable to modify the decrease of enzymatic activity induced by lower work load.     

Coordination of lymphocyte enzyme systems and resistance of mice to the action of staphylococcus toxin]

The activity of acid phosphatase and some dehydrogenases was studied in the blood peripheral lymphocytes of intact mice. Then this mice were given an intraperitoneal injection of LD50 of staphylococcus toxin; in 2 days 42 mice perished and 38 survived. The groups of survived and perished animals differed (the difference was statistically significant) by the extent of coordination of the enzymatic lymphocyte systems: the correlation of enzymatic indices in the survived animals was greater than in the perished ones. The data obtained are discussed from the aspect of a priori intoxication prognosis and the significance of the enzymatic coordination.     

Structural effects on enzymatic degradabilities for poly[(R)-3-hydroxybutyric acid] and its copolymers.

Poly[(R)-3-hydroxybutyric acid] and its copolymers were prepared by biosynthetic and chemosynthetic methods. The films of polyesters were prepared by both the solution-cast and melt-crystallized techniques. The enzymatic degradation of polyester films was carried out at 37 degrees C in an aqueous solution (pH 7.4) of PHB depolymerase from Alcaligenes faecalis. The rate of enzymatic erosion on the solution-cast films increased markedly with an increase in the fraction of second monomer units up to 10-20 mol% to reach a maximum value followed by a decrease in the erosion rate. Analysis of the water-soluble products liberated during the enzymatic degradation of polyester films showed the formation of a mixture of monomers and oligomers of (R)-3HB and hydroxyalkanoic acids units, suggesting that the active site of PHB depolymerase recognizes at least three monomeric units as substrate for the hydrolysis of ester bonds in a polymer chain. The rate of enzymatic erosion of melt-crystallized polyester films decreased with an increase in crystallinity. PHB depolymerase predominantly hydrolyzed the polymer chains in the amorphous phase and subsequently eroded crystalline phase. In addition, the enzymatic degradation of crystalline phase by PHB depolymerase progressed from the edges of crystalline lamellar stacks. The enzymatic erosion rate of crystalline phase in polyester films decreased with an increase in the lamellar thickness.