Enzymatic conversions of polar lipids. Principles, problems and solutions

This text provides a brief overview of the principles of enzymatic lipid conversion and some recent advances in the enzymatic conversion of glycerophospholipids and galactolipids. Lipases and phospholipases are used to exchange fatty acids or the polar group in the lipids. The reactions can be carried out either as hydrolysis–esterification sequences or as one-step transferase reactions. The scope and limitations of the different methods are discussed.     

Ficin-catalyzed asymmetric aldol reactions of heterocyclic ketones with aldehydes

Ficin from fig tree latex displayed a promiscuous activity to catalyze the direct asymmetric aldol reactions of heterocyclic ketones with aromatic aldehydes. Ficin showed good substrate adaptability to different heterocyclic ketones containing nitrogen, oxygen or sulfur. The enantioselectivities up to 81% ee and diastereoselectivities up to 86:14 (anti/syn) were achieved under the optimized reaction conditions.     

Peroxidase-Catalyzed Oxidative Phenolic Coupling of Some Tripeptides Containing Tyrosine and Hydroxyphenylglycine Residues

Several tripeptides containing tyrosine or its mono or dichloro derivative; glycine; and 4-hydroxyphenylglycine or its monochloro derivative were prepared. They were subjected to horseradish peroxidase (HRP)-catalyzed oxidative phenolic coupling. Only the products of intermolecular reaction, primarily C-O and C-C coupled dimers, were obtained.     

Enhanced enzymatic hydrolysis of hydrothermally pretreated empty fruit bunches at high solids loadings by the synergism of hemicellulase and polyethylene glycol

In enzymatic hydrolysis, high lignocellulose loadings are required to obtain high sugar titers. However, the high solids loadings limit enzymatic hydrolysis. In this study, to overcome this limitation, the promoting and synergistic effects of the accessory agents of hemicellulase (i.e., Cellic HTec2) and polyethylene glycol (PEG) 8000 were investigated in the enzymatic hydrolysis of hydrothermally pretreated empty fruit bunches (EFBs). After the optimal addition of Cellic HTec2 and PEG, high enzymatic digestion of the pretreated EFBs was achieved owing to their synergistic effects, even at high solids loadings. For example, the enzymatic digestibility of pretreated EFBs at a 21.7% (w/v) solids loading with 10 FPU of Cellic CTec2/g glucan reached 72.5% when 2.7 mg of Cellic HTec2/g glucan and 62.5 mg of PEG/g glucan were used as the accessory agents. These results suggested that the optimal addition of accessory agents is effective for the enhanced hydrolysis of lignocellulose using even a commercial cellulase preparation.     

Cellulase Hydrolysis of Cotton Cellulose: The Effects of Mechanical Action,Enzyme Concentration and Dyed Substrates

Experiments with dyed and undyed cotton, using different cellulose concentration and with or with out mechanical action showed a high effect of those factors. The process with mechanical action have a higher weight-loss. Dyed cotton with vat and sulfur dyes have almost die same weight-loss than undyed cotton, but on reactive dyed cotton the cellulase hydrolysis is shorter. It was also verified a decrease of hydrolysis extent with an increase of the reactive dye concentration on the cotton substrate. Some synergism due to the enzyme concentration was apparently observed on the changes of the length of the leaving sugars.     

信号肽对亮氨酸脱氢酶在Bacillus subtilis中分泌表达的影响及酶学性质研究

根据信号肽N端电荷数,选择Sec及Tat两种途径的信号肽构建枯草芽孢杆菌穿梭质粒,首次实现Bacillus cereus源亮氨酸脱氢酶基因在Bacillus subtilis中的分泌表达。Tat途径信号肽Pho D促进蛋白质分泌的效果最好,胞外酶活力达20.25U/ml,为不添加信号肽的2.2倍,信号肽N端较多的电荷数,可能有利于多聚体蛋白的分泌。对表达产物进行纯化和酶学性质测定。结果表明,纯酶比酶活为13U/mg;L-Leucine为底物时酶的K_m为6.17mmol/L,V_(max)为14.49μmol/(L·min);底物特异性研究发现,酶与天然底物L-Leucine的亲和性最好,对一些脂肪族氨基酸也有活性,对芳香族氨基酸L-Phenylalanine无活性;酶的最适pH为10.5~12.0,pH稳定范围为5.0~11.0;最适反应温度为55℃;圆二色谱变温扫描酶二级结构变化,α螺旋含量随温度升高逐渐降低;差示扫描微量热技术(DSC)测定酶的解折叠温度(Tm值)为64.13℃,表明该酶具有较好耐热性。     

Strategies for the design and operation of enzymatic reactors for the degradation of highly and poorly soluble recalcitrant compounds

The presence of recalcitrant compounds in both wastewaters and soils is an important environmental problem. Oxidative enzymes from white-rot fungi have been successfully utilised for the in vitro degradation of xenobiotics, such as the azo dye Orange II and the polycyclic aromatic hydrocarbon anthracene (compounds with high and low solubilities, respectively). Two different reactor configurations are proposed: (i) an enzymatic membrane reactor for the treatment of soluble compounds, consisting of a continuous stirred tank reactor coupled to an ultrafiltration membrane to facilitate the retention and recycling of enzyme; and (ii) a two-phase enzymatic reactor for the degradation of poorly soluble compounds, consisting of an immiscible solvent, which contains the contaminant at high concentrations, and the aqueous phase containing the enzyme and cofactors involved in the catalytic cycle. In this paper, factors affecting the design and operation of both systems are discussed, and experimental results concerning the efficiency and stability of the processes are presented.     

Enzymatic Isolation of Archegonium of Pinus Yunnanensis Fr. and Observations on the Proembryo Development

The enzymatic isolation of megagametophyte from fixed and fresh of Pinus yunnanensis Fr. were macerated in a mixture solution of 3% pectinase and 3% cellulase at the 30℃ for 10–13 hours, and the archegonium has been isolated. Materials were stained with A-S, Feulgen and PAS, and following procedure was cleared in the lactophenol. The ventral canal and egg cell which from division of central cell, the functional sperm nucleus becomes pressed against the egg nucleus, and the new cytoplasm round the zygote nucleus were observated under the microscope of usual light. The four proembryonal nuclei divide simultaneously, resulting in eight nuclei arranged in two tiers of four cells each, and following development were formed wall. In the end, the proembryo of six- teen cells were formed. Two methods on enzymatic and dimethyl sulfoxide with potassium hydroxide to isolate the female sexual cells of gymnosperms have been compared, and several problems are discussed.     

酶降解法精制肝素钠

肝素钠在哺乳动物的脏器和体液如：肝、肺、脾及胃肠粘膜等中含量很高．肝素钠作为一种有效的抗凝剂已广泛应用于临床,生物学功能在于阻止血液中凝血酶的作用．其衍生物还具有降血脂、消炎、抗过敏、利尿及调节免疫功能等多种用途．肝素钠属于多糖类化合物,作为溶栓类药...     

Kinetics of Veratryl Alcohol Oxidation by Lignin Peroxidase and in situ Generated Hydrogen Peroxide in an Electrochemical Reactor

The cathodic reduction of oxygen to hydrogen peroxide, the current efficiency for the production of H₂O₂ and the oxidation of veratryl alcohol with an in situ generated hydrogen peroxide‐lignin peroxidase complex were studied in this paper. The complex was prepared by utilizing a novel preparation technique in an electrochemical reactor. The oxidation of veratryl alcohol (VA; 3,4‐dimethoxybenzyl alcohol) was carried out with or without lignin peroxidase under an electric field. The redox properties of veratryl alcohol on a carbon electrode in the presence of lignin peroxidase have been investigated using cyclic voltammetry. The kinetics of veratryl alcohol oxidation in an electrochemical reactor were compared to the oxidation when hydrogen peroxide was supplied externally. Further, the oxidation of veratryl alcohol by lignin peroxidase was optimized in terms of enzyme dosage, pH, and electrical potential. The novel electroenzymatic method was found to be effective using in situ generated hydrogen peroxide for the oxidation of veratryl alcohol by lignin peroxidase.