PURIFICATION AND PROPERTIES OF TRANSGLUTAMINASE FROM STREPTOVERTICILLIUM MOBARAENSE

Transglutaminase (TGase) was separated from the culture broth of an isolated strain of Streptoverticillium mobaraense. The crude enzyme was prepared by centrifugation, ultrafiltration, precipitation by alcohol, centrifugation and freeze‐drying. The yield after these processes was 65–70%. Then the enzyme was purified to homogeneity by chromatography on CM‐cellulose and Sephadex G‐75 on which the yields were about 70% and 80%, respectively; the purified folds reached 2.5–4.7 and 1.08–2.06, respectively. The molecular weight of this TGase was 39,500–40,100 Da by gel filtration chromatography. Optimum enzyme activity was observed in the pH range of 5.0–7.0, and it was maintained stable at 20–40C. The optimal temperature and pH was 52C and 6.0, respectively. At 1 mM and 5 mM metal ion or inhibitors concentration, TGase activity was strongly inhibited by Zn²⁺ and NEM, and not affected obviously by Ba²⁺, Ca²⁺, Co²⁺, Cu²⁺, Fe²⁺, Fe³⁺, Mg²⁺, Mn²⁺, Na⁺ as well as PMSF and EDTA. The effects of these additions on this TGase were compared with those of other microbial TGases.     

贝莱斯芽孢杆菌角蛋白酶的克隆表达、纯化及酶学性质

为了获得更好的羽毛粉降解酶,本文从贝莱斯芽孢杆菌基因组中筛选到一个角蛋白酶基因（baker1）,对其克隆并在大肠杆菌BL21（DE3）中异源表达,最后对重组角蛋白酶（BaKer1）进行了分离纯化、表征及应用研究。实验结果表明,BaKer1的最适反应pH为9.5,最适反应温度为55℃,在中温和碱性条件下具有较好的稳定性。1 mmol/L Ca²⁺对BaKer1有显著地促进作用,5 mmol/L Mn²⁺对BaKer1有轻微地抑制作用,其他金属离子对BaKer1影响不大。PMSF对BaKer1的活性具有显著抑制作用,表明BaKer1是典型的丝氨酸蛋白酶。分别以羽毛粉、偶氮酪蛋白、酪蛋白为底物测定了BaKer1的动力学参数,K_m值分别为5.06、1.09和1.39 mmol/L,k_cat/K_m值分别为1 058.24、2 536.54和3 733.79·(s·mmol/L)。BaKer1处理羽毛粉,能达到酸水解效果的 33.51%,说明它在羽毛粉降解过程中具有潜在的应用价值。     

Antibacterial activity of cranberry juice concentrate on freshness and sensory quality of ready to eat (RTE) foods

Ready-to-eat (RTE) foods are not further treated before consumption in such a way that may significantly reduce the microbial load, therefore the risk of foodborne disease must be considered. In this regard, the use of natural antimicrobial compounds is an interesting method to be considered. On this topic, the antibacterial activity of cranberry juice concentrate (CJC) have been evaluated in vitro and in situ against 3 foodborne pathogenic bacteria. Results showed a high antimicrobial effect with a noticeable inhibition capacity against Escherichia coli O157:H7, Listeria monocytogenes, Salmonella typhimurium. Acid sensitivity studies of bacteria indicated that at the same pH level (pH = 2.4) in presence of organic acid solution (citric and quinic acids), cranberry juice concentrate showed greater antibacterial effects than the acids due to their phenolic compounds. In situ studies showed 2.5, 1.8 and 5 log reduction of E. coli, L. monocytogenes and S. typhimurium, respectively in presence of cranberry juice concentrate, on pre-cut red peppers after 7 days of storage at 4 °C. A total inhibition of L. monocytogenes on fresh cranberry fruits in primary day of storage, was observed. Cranberries treated with CJC also showed a 3 log reduction of S. typhimurium after 4 days of storage at 4 °C. The results suggest that CJC can be an effective preservation, source of natural antibacterial, to protect the RTE foods from foodborne pathogens contamination without effecting on sensorial properties of treated samples and allow to maintain the freshness, sensory and the nutritional quality of RTE foods.     

纯化大肠杆菌表达胆固醇氧化酶的两种亲和分离介质的研究

建立了从重组菌中高效亲和制备胆固醇氧化酶的方法,将一种源自Brevibacterium sp.(DQ345780)的胆固醇氧化酶基因转入Escherichia co●BL21(DE3)表达,,选择核黄素-5’磷酸(FMN)及7-氯-异咯嗪作为亲和配体,构建胆固醇氧化酶亲和制备介质。通过两种介质的一步亲和吸附,获得纯度较高的胆固醇氧化酶样本,蛋白回收率分别为9.4%与9.9%(质量百分比),活性回收率分别为85.2%与93.4%(活性百分比)。使用SDS-PAGE分析,纯化得到蛋白分子量为约50 000,纯度分别为98.0%与97.5%(纯度百分比)。通过静态吸附分析,胆固醇氧化酶相对两种亲和介质的最大理论吸附值分别为71.0与78.5 mg/g介质;解离常数分别为12.8和7.3μg/g介质。     

Preliminary oxidation effect on the conversion of coal in the reaction with methanol and sodium hydroxide

Coals of different rank were oxidized with performic acid to the stage of regenerated humic acids. Both crude and oxidized coals were treated with methanol and sodium hydroxide at 350°C. The final products obtained from the oxidized coals showed higher degree of hydrogenation and solubilization as compared with the corresponding products from crude coals.     

Large permittivity,low loss and defect structure in (Nb,Zn) co-doped SnO2 ceramics studied through a combined experimental and DFT calculational method

Dielectric ceramics with high permittivity and low loss are widely used in electronic components and devices. In this study, (Nb, Zn) co-doped Nb_xZn_ySn_1-x-yO₂ with different doping levels and Nb/Zn ratios was designed to tune the defect structure toward the optimal dielectric performance. The lattice parameters firstly increased from x = y = 0.01 to 0.03 and then decreased, while the oxygen vacancy concentration decreased with doping. The co-doped sample with x = y = 0.02 exhibits stable permittivity up to 800 with an ultra-low loss tanδ ∼0.03 at 40 Hz. DFT calculation showed that the oxygen vacancy was formed with single-Zn doping and co-doping at low doping level, while the hole was generated at higher doping level. The achieved large permittivity and low loss of the sample are related to both Electron-Pinned Defect Dipoles (EPDD) and Hole-Pinned Defect Dipoles (HPDD) effects in the lattice, which was determined by the relative positions of donor and acceptor dopants.     

11α hydroxylation of 16α, 17‐epoxyprogesterone in biphasic ionic liquid/water system by Aspergillus ochraceus

BACKGROUND: The improved efficiency of steroid biotransformation using the biphasic system is generally attributed to the positive effect on the solubility of substrate in aqueous media. A promising alternative for the application of organic solvents in biphasic systems is the use of ionic liquids (ILs). This study aims to investigate the applicability of the biphasic ILs/water system for 11α hydroxylation of 16α, 17‐epoxyprogesterone (HEP) by Aspergillus ochraceus. RESULTS: Of the seven ILs tested, [C₃mim][PF₆] exhibited the best biocompatibility, with markedly improved biotransformation efficiency. In the [C₃mim][PF₆]‐based biphasic system, substrate conversion reached 90% under the condition in which buffer pH, volume ratio of buffer to ILs, cell concentration, and substrate concentration were 4.8, 10/1, 165 g L^?1 and 20 g L^?1, respectively. This is more efficient than that of the monophasic aqueous system. The effects of the cations and anions of these ILs on the 11α hydroxylation of 16α, 17‐epoxyprogesterone (HEP) by A. ochraceus is also discussed. CONCLUSION: The above results showed that IL/water biphasic system improved the efficiency of 11α hydroxylation of 16α, 17‐epoxyprogesterone (HEP) by A. ochraceus, thus suggesting the potential industrial application of ILs‐based biphasic systems for steroid biotransformation. © 2012 Society of Chemical Industry     

Ultrasensitive strain sensors made from metal-coated carbon nanofiller/epoxy composites

A set of resistance-type strain sensors has been fabricated from metal-coated carbon nanofiller (CNF)/epoxy composites. Two nanofillers, i.e., multi-walled carbon nanotubes and vapor growth carbon fibers (VGCFs) with nickel, copper and silver coatings were used. The ultrahigh strain sensitivity was observed in these novel sensors as compared to the sensors made from the CNFs without metal-coating, and conventional strain gauges. In terms of gauge factor, the sensor made of VGCFs with silver coating is estimated to be 155, which is around 80 times higher than that in a metal-foil strain gauge. The possible mechanism responsible for the high sensitivity and its dependence with the networks of the CNFs with and without metal-coating and the geometries of the CNFs were thoroughly investigated.     

Effect of charge trapping species of cupric ions on the photocatalytic oxidation of resorcinol

The photocatalytic oxidation of resorcinol, a potent endocrine disrupting chemical, in oxygenated aqueous suspensions of pure and cupric ions modified Degussa P25 titanium dioxide has been investigated at pH 3.0 ± 0.5. The initial rate of photocatalytic oxidation of resorcinol increased until an optimum dissolved cupric ions concentration was reached at 1.04 mM. At the optimum concentration of cupric ions, the initial rate of photocatalytic mineralisation and degradation of resorcinol was improved by 400%. The observed beneficial effect of cupric ions on the initial rate of resorcinol oxidation could be attributed to the formation of complex and its participation in the photoredox cyclic reaction.Two of the initial oxidation intermediates detected were 1,2,3-trihydroxybenzene and 1,2,4-trihydroxybenzene. These intermediates were formed via hydroxylation of the aromatic ring of resorcinol. Evidences have revealed that 1,2,4-trihydroxybenzene interacted strongly with cupric ions forming copper(II)-trihydroxybenzoate complexes that possessed good adsorption onto TiO₂ surface. These dual-effects help to draw the metal ions closer to the photocatalyst surface and subsequently trigger the electron trapping mechanism by cupric ions. As a result, this improved the charge carriers’ separation. Furthermore, in the presence of oxygen, reoxidation of photoreduced cupric ions occurred and this eliminated the possibility of copper photodeposition, while inducing a photoredox cyclic reaction to regenerate copper species that may potentially act as co-catalyst for the oxidation of 1,2,4-trihydroxybenzene. In contrast, no obvious complex formation was seen between 1,2,3-trihydroxybenzene and cupric ions. This pointed to an intriguing finding which indicates that the positioning of functional group on benzene ring influences the role of cupric ions.     

重组大肠杆菌1,2,4-丁三醇合成途径的平衡优化

1,2,4-丁三醇(1,2,4-butantriol,BT)属于非天然化学品,是军工材料1,2,4-丁三醇三硝酸酯的前体。在重组大肠杆菌中,木糖经脱氢、脱水、脱羧和还原合成BT。但途径各反应不平衡使得中间代谢物积累限制菌体生长和产物合成。本研究首先通过CRISPR/Cas9敲除基因yjh G和yqh D构建无本底表达宿主菌,随后利用不同启动子组合调节BT合成途径中基因xdh、yjh G和yqh D的表达。结果发现,以P_inv表达醇脱氢酶基因yqh D使BT产量达到14.4g/L;以P_tac表达脱氢酶基因xdh和P_{rrn HP1}表达脱水酶基因yjh G的组合方式,BT产量达到15.6g/L,比对照菌株KXW3009分别增加5.9%和14.7%。本研究通过对中间代谢物木糖酸合成和代谢的组合优化,促进了BT的合成,为后续研究提供了借鉴。