绿脓杆菌apoazurin在溶液中存在两种天然构象

如何解释绿脓杆菌apoazurin变性过程的复杂机制是一个有争议的问题.最近的研究表明apoazurin的复杂变性机制可以归结为其天然态存在着至少两种构象.利用内源荧光发射谱和圆二色谱进一步研究了apoazurin的脲变性机制,发现其稳态脲变性符合表观的二态过程,但其动力学为双相过程.在高浓度脲中快反应在几秒钟内完成,而慢反应要经过几个小时.快反应和慢反应的mu值分别为2.24和2.45kJ·mol-1·M-1,去折叠活化能的差值为22kJ·mol-1.时间分辨的荧光发射谱和圆二色谱可以用天然态和完全变性态的谱图通过一个固定的比例参数进行重建.结果表明,过去被广泛接受的存在着变性中间体的机制是不正确的,而apoazurin在天然态存在至少两种构象的假设是合理的.     

复乳法制备红细胞代用品

以单甲氧基聚乙二醇聚乳酸共聚物 [methoxypoly(ethyleneglycol) poly DL lactide,PELA]为膜材 ,用W /O/W的复乳 溶剂扩散法制备了包埋牛血红蛋白 (bovinehemoglobin ,BHb)的微胶囊作为人造红细胞。研究发现复乳过程搅拌分散速率、有机溶剂种类和固化方法对BHb活性有明显影响。当搅拌分散速率小于 90 0 0r/min、以乙酸乙酯为有机相时 ,采用复乳 溶剂扩散法包埋BHb的过程对BHb活性无明显影响。当搅拌分散速率高于 12 0 0 0r/min时 ,BHb活性降低。复乳 溶剂扩散法制备微胶囊过程中固化液体积与微胶囊中BHb活性密切相关 ,通过增大固化液和复乳液的体积比可较好地保持BHb的活性。最后制得了粒径 10 μm左右、包埋率 93%、P50 和Hill系数均接近于天然BHb的微胶囊。     

An Insight into the Active Site of Pseudomonas Fluorecens (P. cepacia) Lipase to Define the Stereochemical Demand for the Transesterification in Organic Solvents

The Pseudomonas fluorescens lipase-catalyzed transesterification of 2-methyl alkanols 1 and the 2-substituted oxiranemethanols 2 with vinyl acetate in organic solvents has been studied and the results discussed in terms of steric and electronic demand within the recently postulated models of the lipase active site.     

Two new 1,4-naphthoquinone derivatives from Impatiens balsamina L. flowers

Two new 1,4-naphthoquinone derivatives, balsaminone D (1), balsaminone E (2) along with two known compounds (3 and 4) were discovered from Impatiens balsamina L. flowers. Their structures were identified with spectroscopic methods including HR–EI–MS, 1D and 2D NMR, as well as the absolute configuration was determined by ECD calculation. In addition, new compounds 1 and 2 with IC₅₀ value of 30.54 and 40.67 μg/mL exhibited better activities against activated t-HSC/Cl-6 cells than positive control Silymarin and Fufang Biejia Ruangan Pian, of which the IC₅₀ value were 202.34 and 231.56 μg/mL, respectively.     

Resolution of N-(2-ethyl-6-methylphenyl) alanine by using microgel beads containing Pseudomonas cepacia lipase

Abstract

Pseudomonas cepacia lipase (PCL) was immobilized in alginate microgel beads by electrostatic dispersion. The high electrical potential applied in the immobilization process could significantly decrease the droplet size. The optimum conditions for lipase immobilization were 2% (w/v) alginate, 100 mM CaCl₂, 8 mg/mL enzyme, 4 kV electrical potential and 200 μm mean bead size. Under these conditions, 78.2 U/g of immobilized PCL activity was obtained with 39.1% retained activity and 57.2% immobilization efficiency. The immobilized PCL (PCL-CA) was subsequently used in the enantioselective hydrolysis of (R, S)-N-(2-ethyl-6-methylphenyl) alanine methyl ester. Although PCL-CA exhibited slightly lower activity than free PCL, it preserved the high enantioselectivity (E-value >?200), which afforded enantiomerically pure (R)-acid (99% e.e._p). Furthermore, PCL-CA exhibited higher thermal stability, storage and medium stability than that of free PCL. Batch-wise operational stability studies demonstrated that PCL-CA retained its initial activity for at least 10 cycles of hydrolysis.     

On the kinetics of phase separation in aqueous two-phase systems

The effect of the tie-line location (phase volume ratio) on the kinetics of phase separation in batch PEG/salt aqueous two-phase systems (ATPS) has been investigated. PEG/sulphate systems with a stability ratio (sr) of 0.34 and 0.37 and relative tie-line lengths in the range 0.1 to 0.6 for a continuous top phase and in the range 0.03 to 0.15 for a continuous bottom phase were used in the batch studies. A continuous settler was designed with three different inlet geometries. Phase separation is much faster when the bottom phase is continuous and in this case the location on the tie-line and the presence or absence of Bacillus subtilis extract makes little difference. When the top phase is continuous the relative sizes of the phases (phase ratio, R, relative distance on tie-line, rd) has an important effect, the larger the top phase (larger R and rd) the slower the phase separation. The presence of Bacillus extract also makes the operation slower which is more marked at the largest values of R (and rd). At the largest volume ratios (R or rd) three different settling regions have been recognised, a region of coalescence, a region of drops moving to the interphase and a region where drops queue at the interphase to coalesce into the large phase. A modified correlation that takes into account the location on the tie-line and thus volume ratio (R) and relative distance (rd) has been proposed and successfully tested. The behavior of batch and continuous systems in the presence and absence of Bacillus subtilis extract in systems with continuous bottom phase was also studied. The settling velocity was lower in the continuous than in the batch systems, and in both cases the initial rate was lower in the presence of Bacillus extract.     

Solvatochromic studies in polyethylene glycol–salt aqueous biphasic systems

The polarities of the co-existing phases of a polyethylene glycol (PEG)-2000–K₃PO₄ aqueous biphasic system (ABS) have been examined using Reichardt’s carboxylated pyridinium-N-phenoxybetaine dye as a probe. Using this probe, the polarities of these phases have been compared to those of conventional solvent extraction systems and micellar systems using values obtained from the literature. In general, these extraction systems are comparable in polarity to rather polar solvents. Data on the free energy of transfer of solvents suggests that this may be due to the failure of the probe to account for the real polarity of the salt-rich phase compared to the polymer-rich phase. Examination of the monophasic region of these systems suggests that the reason for this is that the probe is partitioned to a discreet solvent domain dominated by PEG, even though phase separation of the solution is not observed. The use of linear free energy relationships for the characterization of ABS is briefly discussed.     

Chemoenzymatic synthesis and properties of Schiff bases containing (R)-1-(9-anthryl)ethylamine

Racemic 1-(9-anthryl)ethylamine (10), obtained in 70% overall yield from commercial 9-cyanoanthracene, was kinetically resolved by the Candida antarctica A lipase-catalyzed acetylation with isopropyl acetate as acyl donor, affording (R)-(+)-10 with 95.8% enantiomeric excess (e.e.) (E-value 43.5), which afforded Schiff bases (R)-4 and(R)-8. (1)H-NMR, CD, and MM2 calculations offer a consistent picture of the conformational properties of these potential ligands and an explanation for the limited enhancement of enantioselectivity in cyclopropanation of styrene by their Cu(I) complexes, as compared with previously studied ligands in this series.     

Determination of low levels of poly(ethylene glycol) 400 in plasma and urine by capillary gas chromatography-selected ion-monitoring mass spectrometry after solid-phase extraction

A convenient and sensitive method for the quantitative determination of poly(ethylene glycol) 400 in plasma and urine with capillary gas chromatography-mass spectrometry has been developed. The sample preparation involves solid-phase extraction with subsequent derivatization with heptafluorobutyric anhydride, which proved to give the most stable derivative. The derivatization procedure was optimized using experimental design, and different solid-phase extraction columns were evaluated. The limit of quantitation was 1 μmol/l (0.4 μg/ml) for both plasma and urine.     

圆二色光谱揭示光系统Ⅱ在热处理过程中叶绿素荧光动力学变化的原因

研究了光系统Ⅱ(PSⅡ)在热处理过程中的叶绿素a荧光和圆二色(CD)光谱.在30℃～40℃热处理过程中,PSⅡ的叶绿素荧光参数Fo′保持稳定不变;当温度大于40℃时,Fo'逐渐升高并在55℃达到最大值.在PSⅡ颗粒和富含捕光色素(LHCⅡ)的复合物的热处理过程中,具有超大振幅的CD异常信号出现,并且在40℃时,677 nm的异常CD峰强度达到最大.这些结果暗示在PS Ⅱ颗粒热处理过程中,PSⅡ颗粒中的LHCⅡ的聚集状态和异常CD信号相关,并且也可能是影响Fo′的一个重要因素.     

响应面法优化洋葱假单胞菌产脂肪酶液体发酵工艺

用响应面法对洋葱假单胞菌G-63液体发酵产脂肪酶条件进行了优化。首先运用单因子试验筛选出麦芽糖和豆粉水解液为最适碳源和氮源。在此基础上,通过Plackett-Burman设计试验,对影响产酶条件的11个相关因子进行评估并筛选出具有显著效应的3个因子:橄榄油、豆饼粉水解液以及初始pH值。在用最陡爬坡实验逼近以上3个因子的最大响应区域后,采用响应面分析法,确定出橄榄油、豆粉水解液的最佳浓度和最佳初始pH值分别为4.337%,1.956%和8.38。优化后液体发酵培养基中脂肪酶活力提高到44.39 U/mL,比初始酶活13.45 U/mL提高了3.3倍。