Integrated separation process for isolation and purification of biosuccinic acid

Biotechnologically produced succinic acid has the potential to displace maleic acid and its uses. Therefore, it is of high interest for the chemical, pharmaceutical, and food industry.In addition to optimized production strains and fermentation processes, an efficient separation of succinic acid from the aqueous fermentation broth is indispensable to compete with the current petrochemical production of succinic acid. Isolation and purification of succinic acid from an Escherichia coli fermentation broth were studied with two amine-based reactive extraction systems: (i) trihexylamine in 1-octanol and (ii) diisooctylamine and dihexylamine in a mixture of 1-octanol and 1-hexanol. Back extraction of succinic acid from the organic phase was carried out using an aqueous trimethylamine solution. The trimethylammonium succinate generated after back extraction was split with an evaporation-based crystallization.The focus was on process integration, for example, reuse of the applied amines for extraction and back extraction. It was shown that the maximum trimethylamine concentration for back extraction should not exceed the stoichiometric amount (2 mol trimethylamine/mol the succinic acid in the organic phase) to ensure maximal extraction yields with the reused organic phase in subsequent extractions. Moreover, mixer-settler extraction and back extraction of succinic acid were scaled up from the milliliter- to the liter-scale making use of liquid–liquid centrifuges. The overall yield was 83.5% of the succinic acid from thefermentation supernatant. The final purity of the succinic acid crystals was 99.5%. Organic phase and amines can easily be recycled and reused.     

熊果酸提取工艺的研究进展

简要介绍了熊果酸的生物活性及其应用,综述了熊果酸几种提取工艺的研究进展,探讨了其今后的发展方向。     

Increase of lipid yields from some algae by acid extraction

The amounts of total lipids extracted from some but not all the algae examined were increased significantly by adding HCl to the usual chloroform—methanol extraction mixture. The Yield of phospholipid fraction relative to the glycolipids and neutral lipids increased significantly with acid extraction. Acid extraction also increased the yield of phosphatidyl serine, fatty acids, chlorophyll (or its derivatives) and several unknown compounds.     

我国柠檬酸行业的产业化现状及可持续发展

综述了我国在柠檬酸发酵工艺中菌种选育和原料开发方面取得的进展;在提取工艺中,重点介绍了在万吨级规模应用中获得成功的柠檬酸氢钙法和模拟移动床色谱法。针对我国柠檬酸产业发展中存在的问题提出了相应的策略,以期实现柠檬酸工业的可持续发展。     

正交设计优选橙皮苷提取工艺研究

采用碱-醇提酸沉法提取陈皮中橙皮苷,首先针对提取温度、提取时间、pH值、碱液料液比、醇液料液比、提取次数等因素对提取率的影响进行单因素实验考察,并进一步利用正交实验对橙皮苷的提取工艺进行优化。得到最优浸提条件为:陈皮粉与食用乙醇料液比为1:10、与0.5%NaOH溶液料液比1:10,浸提温度70℃,浸提时间4 h,提取率最高。     

紫锥菊单咖啡酰酒石酸和菊苣酸提取工艺优化

采用正交试验设计,选取乙醇体积浓度百分比、提取温度、提取时间、抗氧化剂用量等因素,优化紫锥菊Echinacea purpurea单咖啡酰酒石酸和菊苣酸的加热回流提取工艺,并考察加入抗氧化剂对提取效果的影响。结果表明,优化的提取条件是以25%乙醇,在80 ℃回流提取90 min。抗氧化剂用量对提取效果影响不显著。优化后的加热回流提取条件对紫锥菊单咖啡酰酒石酸和菊苣酸的提取均适用,提取中无需加入抗氧化剂。     

Separation of lactic acid from fermented broth by reactive extraction

The separation of lactic acid from complex fermentation broth was examined. Liquid–liquid extraction using reversible chemical complexation for reactive extraction was chosen to be the separation method. Over 50% yield of lactic acid was obtained from fermented broth in a single extraction step, when using the tertiary amine as the extractant, 1-dekanol as the diluent and trimethylamine (TMA) as the stripping solution. The effect of complex media on the extraction behaviour has hardly been examined previously.     

青叶胆中齐墩果酸的制备工艺及其纯度测定

研究青叶胆中齐墩果酸的提取工艺及其结构鉴定。用体积分数95%乙醇回流提取2次,每次1.5 h,减压回收乙醇后用水洗涤得沉降物。用YCXY-2号除杂剂处理后,加活性炭脱色30 min 2次,经凝析分离、纯化精制得齐墩果酸产品。采用理化常数测定和薄层色谱、红外光谱、质谱及核磁共振光谱鉴定试样的化学结构,确定实验所得产品为齐墩果酸,而齐墩果酸为青叶胆的主要化学成分。     

Purification of fatty acid ethyl esters by solid-phase extraction and high-performance liquid chromatography

We have developed a two-step method to purify fatty acid ethyl esters (FAEE) using solid-phase extraction (SPE), with a recovery of 70±3% (mean±S.E.M.) as assessed using ethyl oleate as a recovery marker from a standard lipid mixture in hexane. The first step of the SPE procedure involves application of a lipid mixture to an aminopropyl-silica column with simultaneous elution of FAEE and cholesteryl esters from the column with hexane. Gas chromatographic analysis of FAEE without interference from cholesteryl esters may be performed using the eluate from the aminopropyl-silica column, thus eliminating the need for an octadecylsily (ODS) column in this case. The FAEE can then be separated from the cholesteryl esters, if necessary, by chromatography on an ODS column and elution with isopropanol-water (5:1, v/v). Both the aminopropyl-silica and ODS columns were found to be effective for up to four uses. To permit isolation of specific FAEE species following isolation of total FAEE by the two-step SPE method, we have also developed a purification scheme for individaal FAEE by high-performance liquid chromatography (HPLC). Thus, this simple method allows for reproducible isolation of total FAEE by SPE and isolation of individual FAEE species by HPLC.     

Effect of the concentration of DODMAC and 1-decanol on the behavior of reverse micelles in the extraction of amino acids

The concentrations of dioctyldimethyl ammonium chloride (DODMAC) and 1-decanol in isooctane needed to form reverse micelles by phase contact have been determined. The behavior of these reverse micelles in the extraction of aspartic acid, glutamic acid, and threonine was studied by analyzing all of the ionic species in the aqueous phase. The amino acid is extracted from the aqueous phase by exchanging with the Cl(-) counterions of DODMAC in the reverse micelles. The ionic species in the reverse micelles tend toward their undissociated states as the water uptake by the reverse micelles decreases. The effect of 1-decanol on the extraction of the amino acids with two negative charges is due to the change in the water uptake of the reverse micelles. The concentration of DODMAC has no effect on the ion exchange of the amino acid with one negative charge with the Cl(-) counterions of DODMAC in the reverse micelles. Higher molar ratios of decanol to DODMAC favor the selective separation of amino acids with different charge numbers. (c) 1995 John Wiley & Sons, Inc.     

1H NMR based metabolite profiling for optimizing the ethanol extraction of Wolfiporia cocos

Metabolite profiling of Wolfiporia cocos (family: Polyporaceae) had been much advancement in recent days, and its analysis by nuclear magnetic resonance (NMR) spectroscopy has become well established. However, the highly important trait of W. cocos still needs advanced protocols despite some standardization. Partial least squares discriminant analysis (PLS-DA) was used as the multivariate statistical analysis of the ¹H NMR data set. The PLS-DA model was validated, and the key metabolites contributing to the separation in the score plots of different ethanol W. cocos extract. ¹H NMR spectroscopy of W. cocos identified 33 chemically diverse metabolites in D₂O, consisting of 13 amino acids, 11 organic acids 2 sugars, 3 sugar alcohols, 1 nucleoside, and 3 others. Among these metabolites, the levels of tyrosine, proline, methionine, sarcosine, choline, acetoacetate, citrate, 4-aminobutyrate, aspartate, maltose, malate, lysine, xylitol, lactate threonine, leucine, valine, isoleucine, uridine, guanidoacetate, arabitol, mannitol, glucose, and betaine were increased in the 95% ethanol extraction sample compared with the levels in other samples, whereas level of acetate, phenylalanine, alanine, succinate, and fumarate were significantly increased in the 0% ethanol extraction sample. A biological triterpenoid, namely pachymic acid, was detected from different ethanol P. cocos extract using ¹H-NMR spectra were found in CDCl₃. This is the first report to perform the metabolomics profiling of different ethanol W. cocos extract. These researches suggest that W. cocos can be used to obtain substantial amounts of bioactive ingredients for use as potential pharmacological and nutraceuticals agents.     

Method for the extraction of the volatile compound salicylic acid from tobacco leaf material

Salicylic acid (SA) is a signalling compound in plants which is able to induce systemic acquired resistance. In the analysis of SA in plant tissues, the extraction recovery is often very low and variable. This is mainly caused by sublimation of SA, especially during evaporation of organic solvents. Techniques have been designed in order to overcome this problem. In the first part of the extraction procedure, sublimation of SA was prevented by addition of 0.2 M sodium hydroxide. At a later stage of the extraction procedure, sublimation of SA during solvent evaporation was controlled by the addition of a small amount of HPLC eluent. In this way, recoveries in the range of 71-91% for free SA and 65-79% for acid-hydrolysed SA were obtained. Recoveries could be further optimised by the use of an internal standard to correct for volume changes after the addition of the HPLC eluent.     

Acid Lability of Metabolites of 2-Deoxyglucose in Rat Brain: Implications for Estimates of Kinetic Parameters of Deoxyglucose Phosphorylation and Transport Between Blood and Brain

The steady-state brain/plasma distribution ratios of [14C]deoxyglucose ([14C]DG) for hypoglycemic rats previously determined by measurement of DG concentrations in neutralized acid extracts of freeze-blown brain and plasma exceeded those predicted by simulations of kinetics of the DG model. Overestimation of the true size of the precursor pool of [14C]DG for transport and phosphorylation could arise from sequestration of [14C]DG within brain compartments and/or instability of metabolites of [14C]DG and regeneration of free [14C]DG during the experimental period or extraction procedure. In the present study, the concentrations of [14C]DG and glucose were compared in samples of rat brain and plasma extracted in parallel with perchloric acid or 65% ethanol containing phosphate-buffered saline. The concentrations of both hexoses in acid extracts of brain were higher than those in ethanol, whereas hexose contents of plasma were not dependent on the extraction procedure. The magnitude of overestimation of DG content (about 1.2-to fourfold) varied with glucose level and was highest in extracts isolated from hypoglycemic rats; contamination of the [14C]DG fraction with 14C-labeled nonacidic metabolites also contributed to this overestimation. Glucose concentrations in acid extracts of brain exceeded those of the ethanol extracts by less than 40% for normal and hypoglycemic rats.     

制革废弃物中氨基酸的提取和分离

综述了制革废弃物中氨基酸的提取及分离技术的发展状况 ,包括如何从制革厂回收的毛中提取胱氨酸等氨基酸以及如何从皮屑和铬革屑中提取脯氨酸等各种氨基酸的方法     

烟草废料中绿原酸的提取工艺研究

讨论了从烟草废料中提取绿原酸的优势和甲醇、乙醇、丙酮、水等不同溶剂经超声波辅助提取绿原酸的效果：研究结果表明,用体积分数40％的甲醇得到的浸提液中,绿原酸质量浓度为2．11mg／mL,比以水为溶剂时高出近50％.不同浓度的甲醇溶液中,体积分数50％的甲醇提取绿原酸的浓度最高。对树脂的吸附动力学分析表明,大孔树脂CN-101对烟草浸提液中绿原酸的吸附遵循Freundlich等温方程,吸附和解析分离所得的绿原酸收率为87．6％.在超声辅助条件下,利用甲醇等有机溶剂提取烟草中的绿原酸,进而用大孔树脂进行吸附分离的方法可行。     

从发酵液中萃取L-乳酸的工艺

以L-乳酸发酵液为对象,以正丁醇为萃取剂,在pH为2、温度为25℃、n（乳酸）：n（正丁醇）=1：1、乳酸在发酵液中的质量分数为30％时,经3次萃取后,最终的萃取率可达到75．7％。萃取完成后,不需要将乳酸进行反萃,可将所得到的正丁醇-乳酸的混合体系作为底物进行酯化反应,生成乳酸丁酯,从而避开提取纯乳酸的高操作要求。     

白花蛇舌草中熊果酸的提取工艺研究

本研究通过单因素实验,以提取的选择性和提取率为指标,考察了白花蛇舌草中熊果酸的提取过程的几个影响因素;并通过正交实验,对过程参数进行了优化。结果表明,各因素对熊果酸提取效果的影响程度从高到低分别是:提取时间>提取次数>乙醇体积分数>液固比;确定的最适提取条件为:95%乙醇溶液在回流温度下提取两次,每次1.5h,液固比为8:1。     

Synthesis of itaconic acid from agricultural waste using novel Aspergillus niveus

Abstract

Filamentous fungi from the genus Aspergillus are of high importance for the production of organic acids. Itaconic acid (IA) is considered as an important component for the production of synthetic fibers, resin, plastics, rubber, paints, coatings, adhesives, thickeners and binders. Aspergillus niveus MG183809 was isolated from the soil sample (wastewater unit) which was collected from Avadi, Chennai, India. In the present study, itaconic acid was successfully produced by isolated A. niveus by submerged batch fermentation. In the fermentation process, various low-cost substrates like corn starch, wheat flour and sweet potato were used for itaconic acid production. Further, the factor influencing parameters such as substrate concentration and incubation period were optimized. Maximum yield of itaconic acid (15.65?±?1.75?g/L) was achieved by using A. niveus from corn starch at a concentration of 120?g/L after 168?hr (pH 3.0). And also extraction of itaconic acid from the fermentation was performed with 91.96?±?1.57 degree of extraction.     

三种提取乳酸菌中内源性质粒DNA方法的比较

采用3种不同抽提质粒DNA的方法普查了6株不同乳酸菌的内源性质粒DNA,并对3种不同方法进行比较。在相同条件下,琼脂糖凝胶电泳检测的结果显示:效果最佳者为改进的Anderson-Mckay方法,其次为液氮反复冻融方法,最差者为改进的碱裂解方法。为抽提革兰阳性细菌乳酸菌类的内源性质粒DNA提供了行之有效的方法。     

Towards a universally adaptable method for quantitative extraction of high-purity nucleic acids from soil

A universally adaptable protocol for quantitative extraction of high-purity nucleic acids from soil is presented. A major problem regarding the extraction of nucleic acids from soil is the presence of humic substances, which interfere with the extraction process itself and in subsequent analytical manipulations. By the approach described here, the humic compounds are precipitated prior to cell lysis with Al(2)(SO(4))(3), and thus eliminated prior to the nucleic acid extraction. The protocol allows for removing of a considerable content and range of humic acids and should therefore be applicable for a wide spectrum of soil types. Accordingly, reproducible results in analyses of different soil types are made possible, inclusively for quantitative comparisons.