Sequential enzymatic reactions and stability of biomolecules immobilized onto phospholipid polymer nanoparticles

Polymer nanoparticles for sequential enzymatic reactions were prepared by combining a phospholipid polymer shell with a polystyrene core. The active ester groups for the bioconjugation and phospholipid polar groups were incorporated into the phospholipid polymer backbone using a novel active ester monomer and 2-methacryloyloxyethyl phosphorylcholine. For the sequential enzymatic reactions, acetylcholinesterase, choline oxidase, and horseradish peroxidase-labeled IgG were immobilized onto the nanoparticles. As substrates, acetylcholine chloride, choline chloride, and tetramethylbenzidine were added to the nanoparticle suspension, the acetylcholine chloride was converted to choline chloride, the choline chloride was oxidized by choline oxidase, and hydrogen peroxide was then formed as an enzymatic degradation product. The hydrogen peroxide was used for the next enzymatic reaction (oxidized by peroxidase) with tetramethylbenzidine. The sequential enzymatic reactions on the nanoparticles via degradation products (hydrogen peroxide) were significantly higher than that of the enzyme mixture. This result indicated that the diffusion pathway of the enzymatic products and the localization of the immobilized enzyme were important for these reactions. These nanoparticles were capable of facilitating sequential enzymatic reactions.     

Enzymatic deinking of various types of waste paper: Efficiency and characteristics

The aim of the present study was to characterize the enzymatic deinking of various types of waste paper. Studies on the optimization of enzymatic deinking have been performed previously using commercially available enzyme preparations containing cellulase and hemicellulase. The enzymatic deinking of different types of waste paper demonstrated a high efficiency of 86.6% on laser-printed paper, but a low deinking efficiency of 12.9% was obtained with newspaper. All enzymatic treatments significantly improved the drainage rate of the deinked waste paper. Enzymatic deinking increased the tensile index of magazine paper but reduced the tensile index of bubble jet-printed paper, photocopy paper and newspaper. Enzymatic hydrolysis caused a 21.1% reduction in the tear index for bubble jet-printed paper, but a 3.1% increase in the tear index was obtained for laser-printed paper relative to respective blank. In addition, enzymatic hydrolysis increased the burst index by 4.7% relative to blank for laser-printed paper. However, photocopy paper showed the highest reduction (8.3%) in the burst index relative to blank. Taken together, these results suggest that enzymatic hydrolysis is both advantageous and detrimental to the mechanical properties of deinked paper. Thus, the proper regulation of enzymatic hydrolysis is crucial to improve the quality of recycled paper.     

Changes in the structural composition of hardwood kraft pulp during bleaching

The possibility of using xylanase preparations for hydrolyzing hemicelluloses in a non-bleached kraft pulp in order to facilitate its bleaching was studied. The effects of enzymatic preparations of fungal and bacterial origins were examined, and the optimal conditions for xylanase activity were determined. UV spectroscopy demonstrated that the treatment of kraft pulp with enzymatic preparations containing xylanase facilitated the subsequent removal of lignin and increased the brightness by 5%. The effect of enzymatic treatment was retained in the case of peroxide bleaching. The enzymatic preparations studied are promising for the development of chlorine-free pulp bleaching technologies.     

Novel enzymatic properties of DNA-Pt complexes

DNA-Pt complexes have shown novel enzymatic activity as a peroxidase similar to that of horseradish peroxidase in the colorimetric reaction with its substrate. The enzymatic activity of these complexes increased with increasing reaction time and pH in reaction solutions of DNA and K2[PtCl4]. This enhanced enzymatic activity was attributed to the increase in Pt conjugated to DNA in the complex. The enzymatic activity per unit mole of the DNA-Pt complex was significantly higher for complexes prepared with high molecular weight DNA because the enzymatic activity of the complex per repeat unit of DNA was almost constant for these complexes prepared under the same reaction conditions. All the DNA-Pt complexes in this study prepared with different DNA sequences (i.e., [A]20, [G]20, [C]20, [T]20, and [AG]10) exhibited peroxidase enzymatic activity. These complexes showed good thermal stability as compared to native horseradish peroxidase.     

Extracellular activity in Cryptococcus neoformans strains isolated from AIDS patients and from environmental sources

Nineteen Cryptococcus neoformans strains isolated from AIDS patients and 16 from bird droppings were tested for their extracellular activity. Typical enzymatic activity that was different from other medically important yeasts was found. The results obtained may indicate that there are new extracellular enzymatic activities that imply a relationship between C. neoformans and its virulence. A correlation among the different enzymatic activities was also investigated and according to the results obtained no relationship was observed among any of the recorded extracellular enzymatic activities. Research on C. neoformans extracellular enzymatic activity is useful not only to better understand its metabolism but in particular to establish a possible relationship between its virulence and pathogenicity.     

Kinetic and enzymatic adsorption model in a recirculation hollow-fibre bioreactor

A general procedure has been developed to model the behaviour of enzymatic reactions in a membrane bioreactor. This procedure unifies the kinetics of the reaction and the adsorption of the enzyme or enzymatic complexes on the membrane, enabling the selection of the most appropriate kinetic model. The general procedure proposed has been particularized and applied to experimental results obtained with two enzymatic reactions carried out in a hollow-fibre reactor, enzymatic hydrolysis of lactose by β-galactosidase and glucose–fructose isomerization by glucose isomerase. The application of the general model has allowed us to determine the mechanism of the reaction for both kinetic reactions, assuming the adsorption of the enzymatic complex EGa for lactose hydrolysis and the adsorption of the free enzyme onto the membrane for glucose–fructose isomerization.     

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.     

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.

     

乙酸分级预处理甘蔗渣对纤维素酶解性能的影响

为提高甘蔗渣的纤维素酶解性能,采用乙酸脱木素结合碱脱乙酰基的预处理工艺 (Acetoline工艺) 对甘蔗渣进行预处理,考察了乙酸脱木素过程中若干因素对预处理结果的影响,并对预处理后甘蔗渣的纤维素酶解性能进行了研究。结果表明,经过Acetoline预处理后甘蔗渣在7.5％固体含量、15 FPU+10 CBU/g固体的纤维素酶和β-葡萄糖苷酶用量下酶解48 h,酶解聚糖转化率接近80％。与稀酸预处理相比,Acetoline预处理可以得到更高的酶解聚糖转化率。实验结果表明Acetoline工艺是一种可有效提高甘蔗渣纤维素酶解性能的预处理方法。     

Optimization of the unhairing leather processing with enzymes and the evaluation of inter-fibrillary proteins removal: an environment-friendly alternative

In this work is described the isolation of a new proteases-producing strain of Bacillus subtilis, screened from aerobic tannery sludge, to be applied in leather production. The optimization of culture conditions to enhance the proteolytic activity was carried out using central composite design. The enzymatic extract was characterized and the hide unhairing and the inter-fibrillary removal capabilities of the enzymatic extract were evaluated by scanning electron microscopy and by the determination of proteoglycans and glycosaminoglycans. The leather quality obtained with this enzymatic preparation was assessed for possible damages to hide collagen by measuring the amount of hydroxyproline released into the reaction medium. Temperature was the most significant factor for culture conditions optimization. The crude enzymatic extract showed the best values for proteolytic activities at pH 9 and 10, temperature between 37 and 55?°C, and showed good thermal stability up to 45?°C. The treated hides presented few remaining hairs; for the enzymatic process, the removal of inter-fibrillary proteins was approximately fourfold for glycosaminoglycans and sixfold for proteoglycans, when compared with the conventional unhairing process. The enzyme application was successful for hide treatment, suggesting that this enzymatic preparation can be used in an environment-friendly leather production to replace the conventional chemical process.     

Change in composition of structural components of leaf cellulose sulfate during fermentation bleaching by xylanases

The possibility of the use of xylanase preparations for hydrolysing hemicelluloses in a non-bleached kraft pulp in order to facilitate its bleaching was studied. The effects of enzymatic preparations of the fungal and bacterial origins were examined, and the optimal conditions for xylanase activity were determined. UV spectroscopy demonstrated that the treatment of kraft pulp with the enzymatic preparations containing xylanase facilitated the subsequent removal of lignin and increased the brightness by 5%. The effect of enzymatic treatment was retained in the case of peroxide bleaching. The enzymatic preparations studied are promising for the development of chlorine-free pulp bleaching technologies.     

蒸汽爆破处理沙柳原料酶解条件优化

研究蒸汽爆破预处理对沙柳原料酶解效果的影响,通过响应曲面实验设计法优化蒸汽爆破处理沙柳原料的酶解工艺。结果表明,蒸汽爆破预处理沙柳原料的最佳蒸汽爆破处理条件：压力3.5 MPa、维压时间300 s; 蒸汽爆破最佳酶解条件：pH 4.8、温度53.5 ℃、 每克底物酶加量29.8 FPU。在最优条件下,蒸汽爆破处理沙柳原料的酶解率可以达到最大值87.92%,并验证了数学模型的有效性,试验结果表明蒸汽爆破预处理可以有效提高沙柳原料的水解率。     

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.     

Organosolv pretreatment by crude glycerol from oleochemicals industry for enzymatic hydrolysis of wheat straw

In order to defray the cost of biodiesel production, the ensuing work was to further investigate utilization of the crude glycerol (CG) from oleochemicals industry in the atmospheric autocatalytic organosolv pretreatment (AAOP) to enhance enzymatic hydrolysis.

The AAOP–CG enabled wheat straw to achieve with reasonable enzymatic hydrolysis yields, reaching 75% for the wet substrate and 63% for the dried. Lipophilic compounds from the CG formed pitch deposition on the fiber, which was responsible for low delignification (30%) and also troublesome in practical operation. Pitch deposits itself had no significant role on enzymatic hydrolysis. A striking finding of the lignin recondensation and/or lignin–carbohydrate complex helped explain why dried pretreated wheat straw had a low enzymatic hydrolysis yield. The CG was suitable for the AAOP to enhance enzymatic hydrolysis of lignocellulosic biomass. But it was advisable to remove lipophilic compounds from crude glycerol before utilization.     

表面活性剂对木质纤维素酶解产糖的影响研究进展

木质纤维素是生产生物燃料乙醇的主要原料,其含量丰富、绿色环保以及可再生性,因此有效地利用木质纤维素有望解决能源短缺问题。表面活性剂能够有效地促进木质纤维素的酶解反应,通过探讨不同表面活性剂对酶解反应的影响及机理,为实际的酶解过程找到合适表面活性剂提供一定的理论指导。     

The enzymatic activity from the sediment of the Gilau dam reservoir - Cluj county

The enzymological studies on the sediment of the accumulation lake that has the main purpose of supplying drinking water to the city of Cluj-Napoca and the nearby villages, were aimed at the comprehensive understanding of the complex processes that happen in these habitats of special significance. In the sediment samples the following enzymatic activities have been quantitatively determined: phosphatase, actual and potential dehydrogenase, catalase, urease and protease. Non-enzymatic catalytic activity was also measured. Based on the relative values for the enzymatic activities, the enzymatic indicator of the sediment quality (EISQ) was calculated (ranging from 0.1 to 0.7). The enzymatic activities have been qualitatively determined for maltase, saccharase, lactase, cellobiase, amylase, dextranase, levanase, cellulase and inulinase. The correlation between the enzymatic and bacteriologic potential was statistically calculated.     

Isolation of bovine retinal pigment epithelial cells using adhesion to agarose: demonstration of cellular and regional heterogeneity.

The retinal pigment epithelium (RPE) shows cell heterogeneity in morphology and enzymatic activity. Routine isolation procedures for RPE cells may reduce enzymatic activity and prevent the quantification of regional enzymatic differences in vivo. We developed a new technique for the isolation of RPE cells based on adhesion of the cells to agarose. The morphology of the isolated cells resembled that of RPE cells in vivo. The cells were viable in the dye exclusion test and showed a histochemical staining pattern as RPE cells in vivo. With this technique, quantitative regional differences in the enzymatic activities were detected.     

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.     

发酵产丁二酸过程中废弃细胞的循环利用

对厌氧发酵产丁二酸后的废弃细胞进行破壁处理,考察了以细胞水解液作为有机氮源重新用于丁二酸发酵的可行性。比较了超声破碎、盐溶、酶解3种方法破碎细胞获得的水解液作为氮源发酵产丁二酸的效果,结果表明酶解制得的细胞水解液效果最佳。以总氮含量为1.11g/L的酶解液(相当于10g/L酵母膏)作为氮源发酵,丁二酸产量可达42.0g/L,继续增大酶解液用量对耗糖、产酸能力没有显著提高。将细胞酶解液与5g/L酵母膏联用发酵36h后,丁二酸产量达75.5g/L,且丁二酸生产强度为2.10g/(L·h),比使用10g/L酵母膏时提高了66.7%。因此,厌氧发酵产丁二酸结束后的废弃细胞酶解液可以替代原培养基中50%的酵母膏用于发酵。