双水相体系酶法合成L-苯丙氨酸的研究

以天冬氨酸转氨酶为催化剂,开展了双水相体系中苯丙酮酸转氨反应制备L-苯丙氨酸的研究。双水相体系质量组成为聚乙二醇4 000(20%)/Na2HPO4(16%)。苯丙酮酸钠盐、细胞和L-苯丙氨酸在双水相体系中的分配系数分别为8.03,31.7和0.74。当底物浓度为43.4 g/L时,该体系酶法合成L-苯丙氨酸得率为78.1%,比在水溶液体系中提高了27.4%。     

双水相体系萃取精氨酸脱亚胺酶

报道了利用聚乙二醇/硫酸铵双水相体系从自溶NJ402菌粗提酶液中分离纯化精氨酸脱亚氨酶(ADI)的研究结果,为精氨酸脱亚氨酶的分离纯化提供了一种方法。在双水相体系中采用聚乙二醇(PEG)与(NH4)2SO4为组成成分,考察了聚乙二醇(PEG)平均相对分子质量、PEG质量分数、(NH4)2SO4质量分数、pH及NaCl质量分数对精氨酸脱亚氨酶分离纯化效果的影响。最佳双水相体系萃取条件为:聚乙二醇(PEG)平均相对分子质量为1 000,w(PEG1000)=15%,w[(NH4)2SO4]=20%,pH=6.5,室温下从自溶NJ402菌粗提酶液中分离纯化精氨酸脱亚氨酶,纯化倍数达到2.35倍,萃取率达91.1%。     

双水相法回收异戊酰螺旋霉素生产废水中的螺旋霉素

建立了乙醇- K2HPO4双水相体系萃取螺旋霉素的方法,对测得的双水相体系的双节线数据进行拟合,并系统研究了K2HPO4浓度、乙醇浓度、螺旋霉素初始浓度、萃取温度和体系pH对分配系数和萃取率的影响。结果表明,双水相萃取螺旋霉素是自发进行、吸热熵增的过程。当K2HPO4质量分数为20%,乙醇质量分数为16%,萃取温度为25℃,体系pH为9.2~9.5时,分配系数可达36.66以上,萃取率可达97.11%以上。其中,当体系"pH = 9.5" 时,分配系数达到47.52,萃取率达到97.97%。乙醇-K2HPO4双水相体系萃取螺旋霉素的纯化倍数及萃取率高,而且所用成相物质乙醇和K2HPO4可以回收重复使用,避免了二次污染,为处理含螺旋霉素的异戊酰螺旋霉素生产废水提供了新的方法。     

近平滑假丝酵母ATCC7330中羰基还原酶双水相萃取工艺研究

研究了聚乙二醇(PEG)/硫酸铵[(NH_4)_2SO_4]双水相体系萃取近平滑假丝酵母ATCC 7330粗酶液中羰基还原酶的工艺,考察了无机盐种类、PEG分子量及其质量分数、(NH_4)_2SO_4质量分数、萃取温度、萃取时间等因素对羰基还原酶纯化倍数的影响。确定最优的双水相萃取条件为:(NH_4)_2SO_4质量分数15%、PEG1000质量分数19%、萃取温度16℃、萃取时间15 min,在此条件下,羰基还原酶的纯化倍数可达6倍。为近平滑假丝酵母ATCC 7330中羰基还原酶在手性化合物的绿色合成中的应用奠定了基础。     

PEG-(NH_4)_2SO_4双水相萃取法提取壳聚糖酶的研究

采用PEG-(NH4)2SO4双水相体系直接从Bacillussp.LS发酵液上清液中分离壳聚糖酶。研究了体系中PEG分子量、PEG质量分数、(NH4)2SO4质量分数、NaCl质量分数和pH值对壳聚糖酶分配系数及萃取率的影响。结果表明,室温下双水相萃取最佳条件为:PEG600 20%、(NH4)2SO420%、NaCl 0.1%、pH值6.0,在此条件下壳聚糖酶分配系数达5.91,萃取率达88.7%。     

聚乙二醇/硫酸铵双水相体系萃取猪胰蛋白酶

采用聚乙二醇(PEG)/硫酸铵[(NH4)2SO4]双水相体系对猪胰蛋白酶分离进行了研究.通过综合考察酶分配系数、蛋白质分配系数、相比和回收率,探讨了PEG400质量分数,(NH4)2SO4质量分数、NaCl质量分数以及pH值对胰蛋白酶萃取的影响,并通过正交实验进一步优化实验条件,结果表明(NH4)2SO4质量分数和PEG浓度对胰蛋白酶的萃取影响大,在PEG400质量分数为24%、(NH4)2SO4质量分数为21%、pH值为4.2所组成的双水相体系下,可获得酶的高分配系数8.48,提取的胰蛋白酶活力达到1780U/mL.     

双水相体系萃取木瓜蛋白酶的研究

采用聚乙二醇(PEG)/(NH4)2SO4双水相体系对木瓜蛋白酶进行萃取分离,研究了PEG相对分子量、PEG质量分数、(NH4)2SO4质量分数和pH值对木瓜蛋白酶分配系数及酶活力回收率的影响.结果表明,最佳萃取条件为:PEG4000质量分数6%、(NH4)2SO4质量分数18%、pH值6.0,在此条件下,木瓜蛋白酶的...     

双水相萃取藻蓝蛋白的机理

采用荧光光谱的方法对双水相萃取藻蓝蛋白(PC)的机理进行了研究。不同分子量的聚乙二醇(PEG)对PC均有荧光猝灭作用。PEG与PC相互作用的热力学研究结果表明,PEG1000与PC的结合常数大于PEG2000和PEG4000,两者是熵驱动下的疏水作用力结合。不同的无机盐对PEG1000与PC结合的影响不同,其中Na2SO4使PEG1000-PC体系熵增效果最显著。同步荧光光谱的研究表明,PEG1000使PC色氨酸残基微环境的疏水性增强;Na2SO4通过促进PEG1000与PC的结合而使色氨酸残基微环境的疏水性进一步增强。单因素扫描实验的结果表明,PEG1000/Na2SO4体系双水相萃取纯化藻蓝蛋白(PC)的能力最强,符合PEG1000与PC分子相互作用的热力学规律。     

PEG/（NH4）2SO4双水相萃取分离茶氨酸的研究

采用聚乙二醇(PEG)/(NH4)2SO4双水相体系萃取分离生产茶多酚所得废液中的茶氨酸,考察了PEG分子量与含量、硫酸铵含量、pH、温度、加盐(KCl、KBr、KI)、茶氨酸含量对双水相及萃取分离茶氨酸的影响。结果表明,PEG/(NH4)2SO4双水相萃取分离茶氨酸的适宜条件是:PEG平均分子量为4 000,质量分数为10%,硫酸铵质量分数为15%,pH约为6,30℃。在此条件下,茶氨酸的分配系数K1=0.16,蛋白质的分配系数K2=0.28,糖类的分配系数K3=9.8,茶氨酸在下相的萃取率为89.5%,可以将茶多酚废液中的茶氨酸与糖类及其他有颜色的杂质分开。     

蒲公英总黄酮在聚乙二醇-硫酸铵双水相体系中的分配与提取

蒲公英中内含的黄酮类物质具有较高药用价值,用双水相萃取法提取植物中有效成分是新型提取高附加值生物质的有效方法。本文考察了PEG/(NH4)2SO4双水相体系萃取分离蒲公英总黄酮时聚乙二醇相对分子质量、PEG质量分数、(NH4)2SO4质量分数、温度、pH值5个因素对分配行为的影响,并通过正交实验优化了工艺条件。结果表明最佳双水相提取工艺条件为:(NH4)2SO4质量分数18%,PEG1000质量分数23%,pH值5.34,提取温度25℃,NaCl盐的存在与否对萃取影响很小,蒲公英中总黄酮的提取率可达5.47%。因此,使用双水相法提取蒲公英总黄酮是该类提取技术中一种更加绿色环保和高效的方法。     

Impact of polyethylene glycol as additive on the formation and extraction behavior of ionic‐liquid based aqueous two‐phase system

Developing a novel Ionic‐liquid (IL) based aqueous two‐phase system (ATPS) with polyethylene glycol (PEG) as adjuvant for the separation of biomolecules is studied. This original work involves addition of various concentration of PEG (2000, 4000, and 6000 gr/mol) to 1‐butyl‐3‐methylimidazolium acetate+ potassium hydrogen phosphate ATPS to investigate their subsequent effect on phase diagrams and partitioning coefficient of α‐amylase. In another innovative aspect of this work, response surface methodology (RSM) based on three‐variable central composite design was employed to understand the effect of phase forming components on extraction studies of α‐amylase. The addition of small amount of PEG improved the partitioning coefficient of biomolecule. The effective excluded volume theory was applied to correlate the salting‐out ability. As a result, it can be stated that the proposed system can effectively be used in separation and purification studies instead of task specific ILs. © 2015 American Institute of Chemical Engineers AIChE J, 62: 264–274, 2016     

Novel Gas-assisted Three-liquid-phase Extraction System for Simultaneous Separation and Concentration of Anthraquinones in Herbal Extract

abstract Gas-assisted three-liquid-phase extraction (GATE), which has the advantages of both three-liquid-phase extrac-tion and solvent sublation, is a novel separation technique for separation and con...     

Application of Response Surface Methodology for Optimization of Bromelain Extraction in Aqueous Two-Phase System

Extraction of bromelain from pineapple fruit in an aqueous two phase system (ATPS) composed of polyethylene glycol (PEG) 1500 and potassium phosphate has been studied using response surface methodology. The various process variables such as PEG, potassium phosphate and NaCl concentration, and pH were optimized using a central composite rotatable design (CCRD) of response surface methodology (RSM) based on the partition coefficient, % yield, and purification factor of an enzyme. An optimized ATPS composed of 14% (w/w) PEG 1500, 17.66% (w/w) potassium phosphate and 1 mM sodium chloride at pH 7.5 was used to purify bromelain from a pineapple fruit. With this system, a maximum enzyme partition coefficient of 12.62 and %yield of 90.33 in the top PEG-rich phase with a purification factor of 2.4 was predicted. The enzyme partition coefficient, % yield, and purification factor obtained from experimentation are 12.22, 89.65, and 2.8, respectively, in the top PEG phase. The response model is validated by the closeness between the predicted and experimental results.     

Optimization of Rhizopus niveus Lipase Partitioning by an Aqueous Biphasic System

An unconventional aqueous two‐phase system (ATPS) composed of polyethylene glycol (PEG) and sodium carbonate was developed and optimized by employing response surface methodology for separation of Rhizopus niveus lipase. A five‐level central composite design was applied to evaluate the optimal level of three process variables in order to obtain maximum lipase separation. Experimental data were analyzed by regression and a polynomial model was created which was found significant. The maximum partition coefficient was achieved with the system PEG 4000/sodium carbonate. Validation experiments confirmed the high accordance of predicted and experimental results. The optimized ATPS can be applied as a suitable cost‐effective system for lipase extraction.     

椰油丙基酰胺甜菜碱/聚乙二醇水溶液的分相研究

Phase separation behavior of cocamidopropyl betaine/water/polyethylene glycol (PEG) system was studied. The effects of concentration and molecular weight of PEG on the phase separation behavior were investigated. Clouding occurred when the con-centration of PEG was large enough in the betaine aqueous solution, and the concentration of PEG at cloud point decreased with the increase of PEG molecular weight for a constant betaine concentration. The bottom phase was the PEG-rich phase, and the upper phase was the betaine-rich phase. The volumetric ratio of PEG-rich phase to betaine-rich phase, at the same difference between the PEG concentration and the one at the cloud point, Ccp (0.1 g•ml－1), decreased as the PEG molecular weight increased and approached 1 for higher PEG molecular weight (about 20000), which was similar to the typical aqueous two-phase system. This volumetric ratio depended on the initial PEG concentration, but independent of PEG molecular weight. The concentration ratio of betaine to PEG in both phases depended on the Ccp, independent of PEG molecular weight.     

壳聚糖／水溶性高分子共混膜相容性的预测与表征

高分子（聚乙烯醇和聚乙二醇）间的相容性,壳聚糖与极性水溶性高分子在溶液状态时,分子间相互吸引,相容性好,向本体转化而成膜时,CS／PVA体系比CS／PEG体系有更好的本体相容性。原因可归结为PEG不能像PVA一样提供足够多的极性基团与壳聚糖分子形成强烈相互作用,CS与PEG间的氢键强度要弱于CS与PVA间的氢键;另一个方面,PEG结晶能力很强,在由溶液转化为本体而成膜过程中,PEG分子自身之间易于形成较规整的结晶而逐渐使体系发生分相。     

Preparation of monodispersed porous polyacrylamide microspheres via phase separation in microchannels

We report on the formation of polyacrylamide (PAM)/polyethylene glycol (PEG) core/shell droplets in a microchannel via the polymerization-induced phase separation of an acrylamide (AM)/PEG aqueous system. Monodispersed porous PAM microspheres were prepared from the PAM/PEG core/shell droplets, and we examined the effects of experimental parameters on the phase separation process and on the particle size and pore structure of the resulting PAM microspheres. PAM microspheres could be readily obtained with adjustable particle sizes and porosities by altering the PEG and crosslinker contents and by using PEG with different molecular weights. The relation between the swelling value and porosity is correlated.     

ENZYME MASS TRANSFER COEFFICIENT IN AQUEOUS TWO PHASE SYSTEMS: SPRAY EXTRACTION COLUMNS

Fractional dispersed phase hold-up and overall dispersed phase side mass transfer coefficients were measured in 34, 50, 70 and 95 mm i.d. spray columns using an aqueous two phase system. Sodium sulphate-polyethyleneglycol (PEG) (4000)-water formed the aqueous two phase system and amyloglucosidase was used as a solute for the estimation of mass transfer coefficient.

The dispersed phase hold-up (e_D) and volumetric dispersed phase side mass transfer coefficient (K_D) both increased with increasing PEG-rich phase velocity. The e_Dand K_Dawere found to be independent of the column diameter and column height above a certain critical value. An increase in the phase concentration of sodium sulphate and PEG was found to reduce e_Dand K_DEmpirical and semi-empirical correlations for e_D and K_Dahave been developed.     

Two-phase aqueous systems of cetyltrimethylammonium bromide/sodium dodecyl sulfate with and without polyethylene glycol

Aqueous two-phase systems of cetyltrimethylammonium bromide and sodium dodecyl sulfate mixtures without and with polyethylene glycol (PEG) added have been studied at 313.15 K. The results have shown that PEG has a strong effect on the phase diagram of ATPS and microstructures of surfactants aggregates. The addition of PEG leads to the formation of bigger surfactant aggregates, which can be attributed to both the screening effect and the connection effect of PEG. All ATPS, especially those with PEG added, may have potential applications in separation and purification of biomaterials.     

Shear‐Induced Fabrication of Multiple Parallel Alginate Gel Filaments Using Alginate–Polyethylene Glycol Blend

A rigid assembly of alginates is formed in aqueous media primarily via hydrogen bonding between guluronic units. A flow of aqueous alginate solution in a co‐flow capillary can form alginate gel fibers by contact with Ca²⁺ ions in sheath flow. Mixing with polyols [e.g., polyethylene glycol (PEG)] facilitates the shaping of the alginate assembly because PEG disrupts the assembly of the extended alginate chains to instead form alginate–PEG complexes that exhibit shear‐thinning behavior. The shear‐induced fibrous domains of the globular alginate–PEG complexes can be partitioned by a PEG‐rich phase, resulting in multiple parallel alginate gel filaments when the strong ionic‐field‐induced PEG‐rich phase is adjusted and an alginate–PEG complex phase is used as the aqueous two‐phase separation system.