Enzyme-assisted aqueous extraction of rice bran oil

In the present study, rice brain oil was extracted by enzyme-assisted aqueous extraction under optimized aqueous extraction conditions using mixtures of Protizyme^TM (protease; Jaysons Agritech Pvt. Ltd., Mysore, India), Palkodex^TM (α-amylase; Maps India Ltd., Ahmedabad, India), and cellulase (crude cellulase; Central Drug House, Delhi, India). The optimal conditions used were: mixtures of amylase (80 U), protease (368 U), and cellulase (380 U), with 10 g of rice brain in 40 mL distilled water, pH 7.0, temperature 65°C, extraction time 18 h with constant shaking at 80 rpm. Centrifugation of the mixture at 10,000×g for 20 min yielded a 77% recovery of the oil.     

Application of enzyme-assisted aqueous fat extraction to cocoa fat

An enzyme-assisted aqueous fat extraction method was used to extract fat from dry cocoa beans. The beans were dehulled and finely ground. Samples were mixed with water in predetermined ratios. A protease and an enzyme with high activities of both cellulase and hemicellulase were added at 1% of the sample weight, mixed, and incubated for about 6 h at 37°C. The treated samples were then extracted by a water flotation technique. The preextraction treatment with enzymes effectively increased fat yield by about 26%. The observation suggests that the enzyme-assisted aqueous fat extraction method could improve the efficiency of rural fats and oils technologies currently in use in many developing countries.     

Flaking and extrusion as mechanical treatments for enzyme-assisted aqueous extraction of oil from soybeans

Flaking and extruding dehulled soybeans were evaluated as a means of enhancing oil extraction efficiency during enzyme-assisted aqueous processing of soybeans. Cellulase, protease, and their combination were evaluated for effectiveness in achieving high oil extraction recovery from extruded flakes. Aqueous extraction of extruded full-fat soy flakes gave 68% recovery of the total available oil without using enzymes. A 0.5% wt/wt protease treatment after flaking and extruding dehulled soybeans increased oil extraction recovery to 88% of the total available oil. Flaking and extruding enhanced protease hydrolysis of proteins freeing more oil. Treating extruded flakes with cellulase, however, did not enhance oil extraction either alone or in combination with protease. Discrepancies in oil extraction recoveries were encountered when merely considering crude free fat because some oil became bound to denatured protein during extrusion and/or sample drying. Bound fat was unavailable for determination by using the hexane extraction method, but was accounted for by using the acid hydrolysis method for total oil determination. Oil extraction recovery from extruded soybean flakes was affected by oil determination methods, which was not the case for unextruded full-fat soy flour.     

Enzyme-assisted aqueous extraction of shea fat: A rural approach

The use of enzyme-assisted partial hydrolysis as a preextraction treatment in a rural shea fat extraction process to improve upon the extraction rates of the process was explored following an observed possibility in a preliminary investigation. Finely ground shea kernel meal samples were mixed with water in predetermined ratios and heated to inactivate any enzymes present. A crude protease and an enzyme with both hemicellulase and cellulase activities were added and mixed, also in predetermined concentrations. The suspensions were incubated in a waterbath shaker at temperatures ranging from 30 to 45°C for specified periods of time. The treated meal samples were then extracted using an adapted traditional aqueous extraction process. At optimum meal-to-water ratio of 1:2, enzyme concentration of 1%, the natural pH of the meal (about 5.3), and incubation time of 4 h, the enzyme treatment increased the extraction rate from about 40% in the typical traditional system (control) to about 75%, of the total fat content (estimated by the Soxhlet method). The enzyme-treated meal samples were very easy to extract as there was no need to cream or whip out the fat, as is laboriously done in the traditional process. The extracted fat samples had apparently less unsaponifiable matter content and slightly less free fatty acid content and peroxide value, compared to samples from the typical traditional process and, in some cases, the Soxhlet extracted samples. The observations confirmed the results of the preliminary investigations and suggest that the enzyme-assisted preextraction treatment could significantly improve upon the aqueous shea fat extraction process.     

Effect of different parameters on enzyme-assisted extraction of lycopene from tomato processing waste

The present study aims to optimize extraction process of lycopene by using solvents and examine the effect of enzyme treatment on the recovery if any. Different parts of tomato fruits like whole tomato, peel, pulp and industrial waste were screened for lycopene content. The extraction of lycopene was carried out with four different solvents to know their extraction efficiency. Two step extraction processes was followed for enzyme-assisted extraction of lycopene. In the first step waste samples were treated with enzymes i.e. cellulase and pectinase, whereas in second step extraction of lycopene was carried out by solvent. The optimization study with respect to concentrations and incubation time were carried out for both the enzymes. Also the effect of particle size and cooking methods on recovery of lycopene were studied. The results revealed that the tomato peel (417.97 μg/g) contains highest amount of lycopene followed by industry waste (195.74 μg/g), whole tomato (83.85 μg/g) and pulp (47.6 μg/g). When extraction was carried out by tri-mixture (i.e. acetone, ethanol and hexane) got higher recovery of lycopene than the other solvents. Maximum recovery of lycopene was obtained at 1.5% cellulase and 2% pectinase respectively at 4 h of incubation period. The results also indicated that finer the particle size, higher the recovery of lycopene, whereas all cooking methods reduced the recovery of lycopene. The above extraction process will be beneficial to the small scale entrepreneur to improve their socio-economical status.     

双水相萃取技术研究新进展

介绍了双水相萃取技术(ATPE)在生物工程下游工程中的应用现状,综述了近年来ATPE相关研究的进展。为了提高ATPE的选择性和分离效率,不但发展了热分离聚合物体系和表面活性剂混合体系等新型双水相系统(ATPS),而且在组成传统ATPS的聚合物上耦联亲和配基的亲和ATPS也得到发展。与物理场作用、其他分离技术和生物过程的集成克服了单一ATPE的某些不足,是ATPE的发展方向。常规萃取设备和连续化操作技术在ATPE中的应用标志着其工业化日趋成熟,但建立溶质在ATPS中分配的热力学模型和相关理论有待进一步完善。     

Enzyme-assisted aqueous extraction of peanut oil

Enzyme-assisted aqueous extraction of oil from oilseeds is a relatively recent technique. In the present study, peanut oil was extracted under optimized aqueous extraction conditions using Protizyme^™, which is predominantly a mixture of acid, neutral, and alkaline proteases. The optimal conditions were: enzyme concentration of 2.5% (w/w) in 10 g of peanut seeds, pH 4.0, 40°C, and 18 h incubation with constant shaking at 80 rpm. Centrifuging the mixture at 18,000 × g for 20 min separated the oil with a recovery of 86–92%. The merits of this process over existing solvent extraction and/or mechanical pressing methods are discussed.     

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。     

Recovery of flavonoids from grape skins by enzyme-assisted extraction

Grape skin contains large amounts of different flavonoids so it can be used for their recovery. Optimization of enzyme-assisted extraction of flavonoids was conducted using oenological enzyme preparations with respect to enzyme dosage, temperature, extraction time, pH, and enzyme preparation. Optimal conditions were obtained using enzyme preparation Lallzyme EX-V, at the temperature of 45°C, time of 3 h, pH 2.0, and enzyme dosage of 10.52 mg/g. The new optimized extraction method is less expensive, simple, accurate, and selective for the recovery of simple flavonoids. It is based on an environmentally-friendly extraction solvent which may provide a valuable alternative to conventional methods.     

酸团中微量正己烷的离心萃取分离研究

采用离心萃取分离技术,回收己内酰胺水解反应产生的酸性溶液(酸团)中的正己烷;建立工业侧线实验装置,考察了离心萃取机的转速、进料口流量及正己烷含量对分离性能的影响。结果表明:离心萃取分离性能随进料口正己烷含量的增加而增强;进料口流量越大,分离性能越差;设备转速为1 500 r/min时,分离效果最佳;转速为1 500 r/min,进料口流量为0.6 m3/h时,平均分离效率可达40%,分离效果好。     

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。     

双水相体系萃取天冬氨酸转氨酶

研究了不同的双水相体系对天冬氨酸转氨酶的萃取分离效果,对聚乙二醇/无机盐成相系统进行了系统的研究,初步确定PEG1000 Na2HPO4为研究体系,并考察体系pH值、无机盐对双水相体系分配系数和纯化系数的影响。当PEG1000的质量分数为18%、Na2HPO4的质量分数为20%时,双水相体系对天冬氨酸转氨酶的分离系数和纯化系数分别为2 36和2 94。     

双水相萃取黄连生物碱的研究

利用聚乙二醇(PEG)和硫酸铵（(NH4)2SO4）形成的双水相体系萃取黄连生物碱。通过双水相体系的改变,分别对黄连中4种主要生物碱：小檗碱、黄连碱、巴马丁、药根碱在双水相中的分配行为进行研究,并结合紫外分光光度法检测筛选出黄连生物碱最佳萃取工艺。最佳萃取工艺参数：pH = 6,T = 20 ℃,PEG1000质量分数20%,(NH4)2SO4质量分数16%,小檗碱、黄连碱、巴马丁、药根碱的最高萃取率分别为99.79%,,98.04%,99.96%,99.39%。双水相萃取黄连生物碱高效环保可行。     

络合萃取法回收生物油加氢水相中的乙酸

应用络合萃取法对生物油加氢水相中的乙酸进行回收,系统地研究了络合剂和稀释剂的种类、络合剂含量对络合萃取效率的影响,以及化学组分不同的生物油加氢水相的络合提取效果。结果表明,选择三辛胺(TOA)为络合剂,异辛醇为稀释剂,络合剂的用量占萃取剂体系体积分数50%~60%,萃取剂与水相体积比为1时,对生物油加氢水相中乙酸的提取效率最高达到84.4%。而且发现水相中其他的有机酸,如丙酸、丁酸和苯酚也被提取了出来。经络合萃取处理后,生物油加氢水相的化学需氧量(COD)大大降低,降低了后续水处理成本。     

季膦盐离子液体/表面活性剂/盐双水相的性质及萃取效果研究

向水相中加入有机物或盐, 在一定组成范围内, 可以形成密度不同的两相, 即双水相, 目前在生物萃取领域有广泛的应用。离子液体/表面活性剂体系的双水相体系符合绿色化学的要求, 而且目前研究尚不多见。采用离子液体四丁基膦氯化铁盐、十四烷基三丁基膦氯化铁盐（[P4444]FeCl₄、[P44414]FeCl₄）和两种表面活性剂十二烷基硫酸钠、十二烷基苯磺酸钠（SDS、SDBS）及柠檬酸钠混合形成双水相体系, 并绘制了[P4444]FeCl₄（[P44414]FeCl₄）/SDS（SDBS）/盐4个体系的相图, 用粒度测试研究发现, 双水相的上相粒度约为60~80 nm, 下相粒度太小而未能检测出来。使用紫外-可见分光光度法研究了双水相体系对不同性质染料的萃取效果。结果表明, 离子液体的疏水链长度和表面活性剂疏水链上存在苯环与否都对体系中双水相的区域产生影响。4个双水相体系都对水溶性染料有很好的萃取作用, 但是对油溶性染料萃取效果不佳。此项研究促进了离子液体在化学分离领域的开发利用。     

表面活性剂双水相萃取分离氨基酸研究

用溴化十二烷基三乙铵(C12NE)与十二烷基硫钠(SDS)混合体系形成双水相,测定混合摩尔比与分相比例的关系,并以苯丙氨酸为萃取对象研究其在双水相体系中的分配及多级错流萃取效果。实验结果表明,当C12NE与SDS摩尔比为(1.6∶1)—(1.7∶1)时体系有分相,其中当C12NE与SDS摩尔比为1.65∶1时,分相所得上下相体积比为1∶1,且分相时间为10 m in左右,为适合实验条件。双水相体系萃取苯丙氨酸的结果表明,单级萃取率可达80%以上,二级萃取率可达99%以上。     

双水相分离提取胃蛋白酶和琼脂糖的初步研究

以聚乙二醇/硫酸钠双水相体系分离提取琼脂糖和胃蛋白酶.结果表明:琼脂糖的回收率高于90%,胃蛋白酶的回收率约为50%,两者最适宜的分离条件为:pH值为3,Na2SO4浓度为0.4 g·mL-1,PEG10000的浓度为0.002 mol·L-1.     

1,2,4-丁三醇的双水相萃取

对低分子有机溶剂/无机盐双水相体系萃取分离发酵液中1,2,4-丁三醇（1,2,4-butanetriol，BT）进行了深入研究。通过对不同双水相体系的筛选，最终选定无水乙醇/K₂HPO₄双水相体系来萃取分离BT。使用浊点法对以BT为溶剂的无水乙醇/K₂HPO₄双水相体系进行相图的绘制，发现在K₂HPO₄质量分数为19.83%~46.87%范围内均能成相。通过单因素实验，考察双水相体系中无水乙醇/K₂HPO₄质量分数、pH对BT在两相之间分配系数和萃取效率的影响，得到最佳萃取条件为：系统总量10g、pH 9.5，无水乙醇/K₂HPO₄的质量分数为28%/28%，分配系数和萃取效率分别可以达到18.35和95.87%。在最佳萃取条件下，进一步探究了放大实验对体系萃取效率的影响，发现其对分配系数和萃取效率影响较小，体系稳定性高，为工业提取发酵液中BT提供新思路。     

双水相萃取法分离纯化黄酮类化合物的研究进展

综述双水相体系的组成、特点和双水相萃取法分离纯化黄酮类化合物的研究进展。双水相萃取是一项利用不复杂的设备,并在温和条件下进行简单的操作就可获得较高收率和有效成分的新型分离技术,该技术在黄酮类化合物的分离分析中取得了较好的效果,有望成为一种新型的黄酮类化合物的分离纯化方法。