WO2012051774A1 - 一种盐析萃取发酵液中有机酸的方法 - Google Patents

一种盐析萃取发酵液中有机酸的方法 Download PDF

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WO2012051774A1
WO2012051774A1 PCT/CN2010/080287 CN2010080287W WO2012051774A1 WO 2012051774 A1 WO2012051774 A1 WO 2012051774A1 CN 2010080287 W CN2010080287 W CN 2010080287W WO 2012051774 A1 WO2012051774 A1 WO 2012051774A1
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phase
organic acid
fermentation broth
salt
rich
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修志龙
宋志远
魏博超
孙亚琴
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大连理工大学
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
    • C07C51/42—Separation; Purification; Stabilisation; Use of additives
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
    • C07C51/42—Separation; Purification; Stabilisation; Use of additives
    • C07C51/48—Separation; Purification; Stabilisation; Use of additives by liquid-liquid treatment
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
    • C07C51/42—Separation; Purification; Stabilisation; Use of additives
    • C07C51/50—Use of additives, e.g. for stabilisation

Definitions

  • the invention belongs to the technical field of bioengineering, relates to a separation technology of a microbial fermentation liquid, and particularly relates to a method for salt extraction and extraction of an organic acid in a fermentation liquid.
  • Organic acids (formic acid, acetic acid, lactic acid, succinic acid, citric acid, malic acid, fumaric acid, adipic acid, long-chain dibasic acid, etc., in which long-chain dibasic acid is an organic acid of more than ten carbons) as a promising chemical Raw materials and intermediates are widely used in many fields.
  • acetic acid is mainly used in the manufacture of polyvinyl acetate and cellulose acetate (also known as acetate), and can also be used to synthesize various derivatives such as methyl acetate, ethyl acetate, propyl acetate, butyl acetate, and the like.
  • acetic acid is also an excellent solvent for the paint and paint industry; lactic acid can be used as a preservative, seasoning and sour agent, while lactic acid-based polylactic acid PLA can be widely used in medicine, plastics, cosmetics and agriculture; succinic acid is An important C 4 platform compound, which can be derivatized to produce bulk chemicals such as 1,4-butanediol, tetrahydrofuran, ⁇ -butyrolactone, etc. Succinic acid is also widely used in medicine, food, pesticides, perfumes, rubber, protective coatings. In industries such as dyes and photographic materials. Long-chain dibasic acids are important fine chemical intermediates that can synthesize a range of high value-added specialty chemicals such as perfumes, specialty nylons, and polyamide hot melt adhesives.
  • the synthesis methods of organic acids are mainly chemical synthesis methods and microbial fermentation methods.
  • the chemical synthesis method has low efficiency, high cost, serious environmental pollution and low product purity, and the microbial fermentation method is widely concerned because of abundant raw materials, high yield and low cost.
  • it is relatively difficult to separate the organic acid from the fermentation broth.
  • the organic acid fermentation broth in addition to the target product organic acid, it also contains various components such as bacteria, protein, and residual sugar.
  • Traditional organic acid extraction methods mainly include calcium salt method and organic solvent solution. - Liquid extraction, ion exchange adsorption, and electrodialysis.
  • Calcium salt method is also called lime -
  • the sulfuric acid method is a traditional method for extracting organic acids from fermentation broth, but the method has low yield, large unit operation loss, and generates a large amount of solid waste calcium sulfate, causing serious environmental pollution in environmental problems. Today, this method is obviously not in line with environmental requirements.
  • Organic solvent solution - The liquid extraction method omits the steps of neutralization and acid hydrolysis of the calcium salt method, and does not produce calcium sulfate, which saves raw materials and reduces environmental pollution. Yushan Hao et al.
  • CN 101525285 Separating the acetic acid in the biological hydrogen fermentation broth by bipolar membrane electrodialysis. Although environmentally friendly, the requirements for the treatment liquid are high, and it is necessary to remove the bacteria, proteins, macromolecular substances, and high-priced ions in the fermentation liquid.
  • Yao Zhongren Publication number CN 1887843A
  • Wu Hao et al. Public No.
  • CN101475464A Separation and extraction of succinic acid from the succinic acid fermentation broth by nanofiltration method.
  • the method is carried out by two ultrafiltrations and nanofiltration, and concentrated under reduced pressure to obtain succinic acid after cooling and crystallization, but the nanofiltration membrane needs periodic cleaning. A large amount of waste liquid will be generated, causing certain environmental problems.
  • there is a common problem in the current method for separating and extracting organic acids from organic acid fermentation broths that is, the fermentation broth needs to be separated by removing the cells and proteins by filtration or centrifugation.
  • the 1,3-propanediol and 2,3-butanediol in the fermentation broth are extracted by a novel aqueous two-phase system (Publication No. CN 101012151A; Publication No. CN 101531652A).
  • the salting-out effect has obvious effects on the extraction of hydrophilic organic solvents, and also has a significant improvement effect on traditional organic extraction.
  • Salting out and extraction combine to form a novel salting-out extraction technique, and the new aqueous two-phase extraction is A form of salting out extraction. So far, the separation and extraction of organic acids in the fermentation broth by the salting-out extraction system has not been reported in the literature.
  • the salting-out extraction technology has high partition coefficient, high yield, low solvent dosage, low cost, mild conditions, weak corrosiveness to equipment, and the inorganic salts and organic salts used can be recovered, and multi-stage salting-out extraction or application to fermentation. in.
  • the solvent used in the salting out extraction has a low boiling point and is advantageous for subsequent purification steps.
  • the salting-out extraction technology can directly separate and extract organic acids from the fermentation broth, eliminating the step of filtering and removing bacteria and proteins, simplifying the operation steps, reducing costs and reducing losses.
  • the invention utilizes the salting-out extraction technology to extract the organic acid in the fermentation liquid, and provides a new idea for extracting and separating the organic acid directly from the fermentation liquid, so that the fermentation method for producing organic acid has a broader industrial prospect.
  • the invention aims at the problems of low efficiency, high cost, difficulty in removing bacteria, protein and glucose, etc. from the extraction of the target product from the organic acid fermentation liquid, and proposes a new method for extracting the organic acid in the fermentation liquid by using salt extraction extraction technology. method.
  • the fermentation broth may be a fermentation simulant, or may be a supernatant for removing bacteria and proteins by filtration or an untreated fermentation stock solution, and an organic acid in the fermentation broth.
  • concentration is 20 ⁇ 170g/L and the pH is 2 ⁇ 10.
  • the upper phase is the solvent phase or the extraction phase rich in organic acid
  • the lower phase is the salt-rich phase or the raffinate phase
  • the microbial cells and proteins form a solid phase layer between the two phases.
  • the upper phase is a solvent phase
  • the middle phase is rich in an organic acid phase
  • the lower phase is a salt phase rich in other components.
  • Soluble inorganic salt is Sodium chloride, ammonium sulfate, sodium sulfate, sodium carbonate, potassium carbonate, sodium phosphate, potassium phosphate, disodium hydrogen phosphate, dipotassium hydrogen phosphate, sodium dihydrogen phosphate or potassium dihydrogen phosphate.
  • the soluble organic salt is sodium acetate, potassium acetate, sodium citrate, potassium citrate, sodium oxalate or potassium oxalate.
  • the extractant is methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, acetone, ethylene glycol, diethyl ether, methyl acetate or ethyl acetate.
  • the invention solves the problem that the separation of the organic acid product produced by the fermentation method is difficult, and the salt extraction extraction system has lower viscosity, faster phase separation and lower cost than the conventional high-polymer aqueous two-phase system, and is very advantageous for industrial operation.
  • the salting-out extraction system can directly treat the fermentation liquid, and remove the bacteria, proteins and polysaccharides in the fermentation liquid while separating and extracting, and solve the problems that the ultra-filtration removal power consumption is large and the membrane needs to be cleaned.
  • the target product organic acid is partitioned into the lower boiling organic solvent phase, which is very advantageous for subsequent separation and purification.
  • Example 1 Salting out extraction separation of formic acid
  • a formic acid fermentation simulant at a concentration of 20 g / L was prepared. Take 4.7g of simulated solution and add 2.8g of dipotassium hydrogen phosphate and 2.5g Methanol, stirred uniformly, and allowed to stand at room temperature to form two phases, the upper phase is an alcohol phase rich in formic acid, and the lower phase is a salt phase rich in other components.
  • the partition coefficient of formic acid was 34.8 and the recovery was 98.6%.
  • Example 2 Salting out extraction and separation of lactic acid
  • the lactic acid fermentation broth was filtered through a hollow fiber membrane to obtain a clarified filtrate having a lactic acid concentration of 170 g / L.
  • a clarified filtrate having a lactic acid concentration of 170 g / L.
  • Dipotassium hydrogen phosphate and 3 g of ethanol are stirred uniformly and allowed to stand at room temperature to form two phases.
  • the upper phase is an alcohol phase rich in lactic acid, and the lower phase is a salt phase rich in other components.
  • the partition coefficient of lactic acid is 7.0, and the recovery rate is 98.7%. .
  • lactic acid fermentation broth Take 5.6g of lactic acid fermentation broth at a concentration of 170g / L, add 1.4g of dipotassium hydrogen phosphate and 3g Ethanol, stirring evenly, standing at room temperature, the upper phase is a lactic acid-rich alcohol phase, the lower phase is a salt phase rich in other components, and the bacteria and protein are in two phases.
  • the partition coefficient of lactic acid is 11.2, and the recovery rate is 98.7%.
  • the removal rates of protein and bacteria were 83.9% and 100%, respectively.
  • a lactic acid fermentation simulating solution having a concentration of 170 g/L was prepared. Take 4.7g of simulated solution and add 2.8g of dipotassium hydrogen phosphate and 2.5g Methanol, stirred uniformly, and allowed to stand at room temperature to form two phases, the upper phase is a lactic acid-rich alcohol phase, and the lower phase is a salt phase rich in other components.
  • the partition coefficient of lactic acid was 37.2 and the recovery was 98.7%.
  • a lactic acid fermentation simulating solution having a concentration of 170 g/L was prepared. Take 5.0g of simulated solution, add 2.5g potassium phosphate and 2.5g Ethanol, stirred uniformly, and allowed to stand at room temperature to form two phases, the upper phase is a lactic acid-rich alcohol phase, and the lower phase is a salt phase rich in other components.
  • the partition coefficient of lactic acid was 2.19 and the recovery was 83.2%.
  • a lactic acid fermentation simulating solution having a concentration of 170 g/L was prepared. Take 5.0g of simulated solution, add 2.5g ammonium sulfate and 2.5g Methanol, stirred uniformly, and allowed to stand at room temperature to form two phases, the upper phase is a lactic acid-rich alcohol phase, and the lower phase is a salt phase rich in other components.
  • the partition coefficient of lactic acid was 2.06, and the recovery rate was 77.6%.
  • Example 3 Salting out extraction separation of acetic acid
  • the acetic acid fermentation broth was filtered through a hollow fiber membrane to obtain a clear filtrate having an acetic acid concentration of 40 g / L. Take 5.6g of filtrate and add 1.4g Dipotassium hydrogen phosphate and 3 g of ethanol are stirred uniformly and allowed to stand at room temperature to form two phases.
  • the upper phase is an alcohol phase rich in acetic acid, and the lower phase is a salt phase rich in other components.
  • the partition coefficient of acetic acid was 5.4 and the recovery was 98.8%. .
  • An acetic acid fermentation simulant at a concentration of 40 g/L was prepared. Take 4.9g of simulated liquid and add 2.6g of dipotassium hydrogen phosphate and 2.5g Methanol, stirred uniformly, and allowed to stand at room temperature to form two phases, the upper phase is an alcohol phase rich in acetic acid, and the lower phase is a salt phase rich in other components.
  • the partition coefficient of acetic acid was 30.6 and the recovery was 98.6%.
  • the succinic acid fermentation broth was filtered through a hollow fiber membrane to obtain a clear filtrate having a succinic acid concentration of 45 g/L, which was added to 6.5 g of the clarified fermentation broth.
  • 1.5 g of dipotassium hydrogen phosphate and 2 g of ethanol were stirred uniformly and allowed to stand at room temperature to form two phases, the upper phase being an alcohol phase rich in succinic acid, and the lower phase being a salt phase rich in other components.
  • the partition coefficient of succinic acid is 6.4 and the recovery is 92.3%.
  • the dodecyl dibasic acid fermentation broth was filtered through a hollow fiber membrane to obtain a clear filtrate, and the concentration of the dodecane dibasic acid was 57 g/L.
  • concentration of the dodecane dibasic acid was 57 g/L.
  • To 5.5g Add 2.0g of dipotassium hydrogen phosphate and 2.5g to the clarified fermentation broth
  • the ethanol is stirred uniformly and allowed to stand at room temperature to form two phases.
  • the upper phase is an alcohol phase rich in a dodecane dibasic acid, and the lower phase is a salt phase rich in other components.
  • the partition coefficient of dodecane dibasic acid is 37.5 and the recovery rate is 98.7%. .
  • a dodecaine dibasic acid fermentation simulant with a concentration of 57 g / L was prepared. Take 5.5g of simulated liquid and add 2.0g of dipotassium hydrogen phosphate and 2.5 g of n-butanol, stirred evenly, and allowed to stand at room temperature to form a three-phase, the upper phase is an alcohol phase, the middle phase is rich in a twelve carbon dibasic acid phase, and the lower phase is a salt phase rich in other components. The recovery of dodecane dibasic acid was 88.6%.

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Description

一种盐析萃取发酵液中有机酸的方法
技术领域
本发明属于生物工程技术领域，涉及到微生物发酵液的分离技术，特别涉及一种盐析萃取发酵液中有机酸的方法。
背景技术
有机酸（甲酸、乙酸、乳酸、琥珀酸、柠檬酸、苹果酸、延胡索酸、己二酸、长链二元酸等，其中长链二元酸为十碳以上有机酸）作为生产前景广阔的化工原料和中间体，在很多领域都有着广泛的应用。其中乙酸主要用于制造聚乙酸乙烯酯和纤维素乙酸酯（又称醋酸纤维），也可用于合成各种衍生物，如乙酸甲酯、乙酸乙酯、乙酸丙酯、乙酸丁酯等，同时乙酸也是涂料和油漆工业的极好溶剂；乳酸可用作防腐剂、调味料及酸味剂，而以乳酸为单体的聚乳酸 PLA 可广泛应用于医药、塑料、化妆品及农业中；琥珀酸是一种重要的 C4 平台化合物，可以衍化生成 1,4- 丁二醇、四氢呋喃、 γ- 丁内酯等大宗化学品，琥珀酸也广泛用于医药、食品、农药、香料、橡胶、防护涂料、染料和照相材料等工业中。长链二元酸是重要的精细化工中间体，可以合成香料、特种尼龙、聚酰胺热熔胶等一系列高附加值的特殊化学品。
有机酸的合成方法主要为化学合成法和微生物发酵法。化学合成法效率低、成本高、环境污染严重且产品纯度低，而微生物发酵法则原料丰富、产量高、成本低，因此受到广泛关注。但从发酵液中分离有机酸相对较困难，在有机酸发酵液中，除了目标产物有机酸外，还含有菌体、蛋白质、残糖等多种成分。传统的有机酸提取方法主要有钙盐法、有机溶剂液 - 液萃取法、离子交换吸附法和电渗析法等。钙盐法也称石灰 - 硫酸法，是一种传统的从发酵液中提取有机酸的方法，但此法收率低、单元操作损失多，而且会产生大量的固体废弃物硫酸钙，造成严重的环境污染，在环境问题日益严重的今天，这种方法显然不符合环保的要求。有机溶剂液 - 液萃取法省略了钙盐法的中和与酸解等步骤，而且不会产生硫酸钙，节省了原料，也减少了环境污染。宇山浩等人（公开号 CN 101410526A ）将 pH4.8 或以下的乳酸发酵液用选自甲苯、二甲苯、已基苯、甲醇、乙醇、丙醇、丁醇和溶剂油的至少一种进行提取分离得到乳酸。刘纯等人（公开号 CN 1070394A ）应用甲基异丁基酮等有机溶剂精制十二碳二元酸。但其萃取过程中用到了大量的毒性有机溶剂，残留溶剂的毒性会对产品质量产生不良影响，而且该法不适合高离子强度的发酵液体系，此外还有溶剂回收问题。孙志浩等人 （公开号 CN 101348429 ）采用阳离子树脂交换提取发酵液中琥珀酸，但琥珀酸发酵液中的大量杂质离子会导致树脂污染严重，交换容量迅速下降，而且树脂的频繁再生会产生大量废液，造成严重的环境污染，使其应用受到限制。唐婧等人（公开号 CN 101525285 ）应用双极膜电渗析技术分离生物制氢发酵液中的乙酸。虽然环保，但对处理液的要求较高，需要除去发酵液中的菌体、蛋白质、大分子物质及高价离子。此外，姚中等人（公开号 CN 1887843A ）利用膜分离、活性炭脱色及结晶技术从厌氧发酵法制备的发酵液中分离提取琥珀酸，但膜的清洗加大了劳动强度，而且 50 ℃ 脱色及 65 ℃ 和 -0.08~-0.1MPa 下的减压蒸发浓缩，会导致大量的能耗，价格昂贵的活性炭也增加了提取成本，而且 发酵液酸化时会产生大量硫酸盐，极可能造成硫酸盐超标及收率下降 。吴昊等人（公开号 CN101475464A ）利用纳滤法从琥珀酸发酵液中分离提取琥珀酸，本方法通过两次超滤和纳滤后，减压蒸发浓缩，冷却结晶后得到琥珀酸，但纳滤膜需要周期性的清洗，会产生大量的废液，造成一定的环境问题。此外，目前所报道的从有机酸发酵液中分离提取有机酸的方法都存在一个共同问题，即发酵液都需要通过过滤或离心的方法除去菌体和蛋白之后再进行分离。
近几年一些研究者在进行溶剂析出结晶研究时发现，在适当温度条件下，如果体系中无机盐、有机溶剂和水的浓度适当，这类体系中的盐可能不会析出结晶，而是出现液液分相现象。如果所用有机溶剂是亲水的低分子物质，如甲醇、乙醇、丙酮等，就可以形成一种新型的双水相萃取体系。与传统的高分子聚合物双水相体系相比，这种新型的双水相体系黏度低、分相快、不易乳化且成本低。尽管这方面研究在国内外刚刚起步，但已表现出良好的分离性能， 如 用新型双水相体系萃取发酵液中 1,3- 丙二醇和 2,3- 丁二醇 （ 公开号 CN 101012151A ； 公开号 CN 101531652A ）。 实际上，盐析效应既对亲水性有机溶剂的萃取有明显效果，对传统的有机萃取也有显著的改善效果，盐析与萃取结合形成新型的盐析萃取技术，而新型双水相萃取是盐析萃取的一种形式。到目前为止，盐析萃取体系分离提取发酵液中的有机酸尚未见文献报道。盐析萃取技术分配系数高、收率高、溶剂用量少、成本低、条件温和，对设备的腐蚀性弱，而且所用无机盐和有机盐可回收，进行多级盐析萃取或应用于发酵中。盐析萃取中所使用的溶剂沸点低，有利于后续的纯化步骤。最值得关注的是盐析萃取技术可以直接从发酵液中分离提取有机酸，免除了过滤去除菌体和蛋白这一步，简化了操作步骤，降低了成本，减少了损失。 本发明利用盐析萃取技术萃取发酵液中的有机酸，为直接从发酵液中提取分离有机酸提供了一种新思路，使发酵法生产有机酸有更广阔的工业前景。
发明内容
本发明针对目前从有机酸发酵液中提取目标产物存在的效率低，成本高，菌体、蛋白质及葡萄糖等去除困难等问题，提出利用盐析萃取技术来提取分离发酵液中的有机酸的新方法。
本发明的技术方案：
发酵液可以是发酵模拟液，也可以是 经过滤除去菌体和蛋白的清液 或者是未经处理的发酵原液， 发酵液中有机酸 浓度为 20~170g/L ， pH 值为 2~10 。 向发酵液中添加 一种或二种以上 可溶性无机盐或有 机盐（固体或浓溶液），以及单一或复合萃取剂， 搅拌均匀后于室温下静置分相 1~8h ，其中 盐质量百分数为 15~35% ，萃取剂的质量百分数为 15~35% 。当盐析萃取形成两相时，上相即为富含有机酸的溶剂相或萃取相，下相为富盐相或萃余相， 菌体和蛋白在两相之间形成固相层 ；当形成三相时，上相为溶剂相，中相为富含有机酸相，下相为富含其他成分的盐相。
可溶性无机盐为 氯化钠、硫酸铵、硫酸钠、碳酸钠、碳酸钾、磷酸钠、磷酸钾、磷酸氢二钠、磷酸氢二钾、磷酸二氢钠或磷酸二氢钾等。可溶性有机盐为醋酸钠、醋酸钾、柠檬酸钠、柠蒙酸钾、草酸钠或草酸钾。 萃取剂为甲醇、乙醇、正丙醇、异丙醇、正丁醇、异丁醇、丙酮、乙二醇、乙醚、乙酸甲酯或乙酸乙酯。
本发明的效果和益处：
本发明解决了发酵法生产有机酸产物分离困难的问题，而且较传统的高聚物双水相体系相比，这种盐析萃取体系粘度低、分相快、成本低，非常利于工业化操作。此外，这种盐析萃取体系可以直接处理发酵液，在分离萃取的同时去除发酵液中的菌体、蛋白、多糖，解决了超滤除杂动力消耗较大、需要对膜进行清洗等问题。另外，目标产物有机酸分配于沸点较低的有机溶剂相中，非常有利于后续的分离提纯。
具体实施方式
以下结合技术方案详细叙述本发明的具体实施方式。
实施例 1 ：甲酸的盐析萃取分离
配制浓度为 20g /L 的甲酸发酵模拟液。取 4.7g 模拟液，加入 2.8g 磷酸氢二钾和 2.5g 甲醇，搅拌均匀，室温静置，形成两相，上相为富含甲酸的醇相，下相为富含其它成分的盐相。甲酸的分配系数为 34.8 ，回收率为 98.6% 。
实施例 2 ：乳酸的盐析萃取分离
将乳酸发酵液经中空纤维膜过滤，得澄清滤液，乳酸浓度为 170g /L 。取 5.6g 滤液，加入 1.4g 磷酸氢二钾和 3g 乙醇，搅拌均匀，室温静置，形成两相，上相为富含乳酸的醇相，下相为富含其它成分的盐相。乳酸的分配系数为 7.0 ，回收率为 98.7% 。
取浓度为 170g /L 的乳酸发酵液 5.6g ，加入 1.4g 磷酸氢二钾和 3g 乙醇，搅拌均匀，室温静置分相，上相为富含乳酸的醇相，下相为富含其它成分的盐相，菌体和蛋白处于两相之间。乳酸的分配系数 11.2 ，回收率为 98.7% ，蛋白和菌体的去除率分别为 83.9% 和 100% 。
配制浓度为 170g/L 的乳酸发酵模拟液。取 4.7g 模拟液，加入 2.8g 磷酸氢二钾和 2.5g 甲醇，搅拌均匀，室温静置，形成两相，上相为富含乳酸的醇相，下相为富含其它成分的盐相。乳酸的分配系数为 37.2 ，回收率为 98.7% 。
配制浓度为 170g/L 的乳酸发酵模拟液。取 5.0g 模拟液，加入 2.5g 磷酸钾和 2.5g 乙醇，搅拌均匀，室温静置，形成两相，上相为富含乳酸的醇相，下相为富含其它成分的盐相。乳酸的分配系数为 2.19 ，回收率为 83.2% 。
配制浓度为 170g/L 的乳酸发酵模拟液。取 5.0g 模拟液，加入 2.5g 硫酸铵和 2.5g 甲醇，搅拌均匀，室温静置，形成两相，上相为富含乳酸的醇相，下相为富含其它成分的盐相。乳酸的分配系数为 2.06 ，回收率为 77.6% 。
实施例 3 ：乙酸的盐析萃取分离
将乙酸发酵液经中空纤维膜过滤，得澄清滤液，乙酸浓度为 40g /L 。取 5.6g 滤液，加入 1.4g 磷酸氢二钾和 3g 乙醇，搅拌均匀，室温静置，形成两相，上相为富含乙酸的醇相，下相为富含其它成分的盐相。乙酸的分配系数为 5.4 ，回收率为 98.8% 。
取浓度为 40g /L 的乙酸发酵液 5.6g ，加入 1.4g 磷酸氢二钾和 3g 乙醇，乙酸的分配系数 6.9 ，回收率为 98.8% ，蛋白和菌体的去除率分别为 81.9% 和 100% 。
配制浓度为 40g/L 的乙酸发酵模拟液。取 4.9g 模拟液，加入 2.6g 磷酸氢二钾和 2.5g 甲醇，搅拌均匀，室温静置，形成两相，上相为富含乙酸的醇相，下相为富含其它成分的盐相。乙酸的分配系数为 30.6 ，回收率为 98.6% 。
实施例 4 ：琥珀酸的盐析萃取分离
将琥珀酸发酵液经中空纤维膜过滤，得澄清滤液，琥珀酸浓度为 45g/L ，向 6.5g 的澄清发酵液中加入 1.5g 磷酸氢二钾和 2g 乙醇，搅拌均匀，室温静置，形成两相，上相为富含琥珀酸的醇相，下相为富含其它成分的盐相。琥珀酸的分配系数为 6.4 ，回收率为 92.3% 。
取浓度为 45g/L 的琥珀酸发酵液 5.6g ，加入 1.5g 磷酸氢二钾和 2g 乙醇，琥珀酸的分配系数 8.2 ，回收率为 98.7% ，蛋白和菌体的去除率分别为 85.9% 和 100% 。
实施例 5 ：十二碳二元酸的萃取分离
将十二碳二元酸发酵液经中空纤维膜过滤，得澄清滤液，十二碳二元酸浓度为 57 g/L 。向 5.5g 的澄清发酵液中加入 2.0g 磷酸氢二钾和 2.5g 乙醇，搅拌均匀，室温静置，形成两相，上相为富含十二碳二元酸的醇相，下相为富含其它成分的盐相。十二碳二元酸的分配系数为 37.5 ，回收率为 98.7% 。
取浓度为 57 g /L 的十二碳二元酸发酵液 5.5g ，加入 2.0g 磷酸氢二钾和 2.5g 乙醇，十二碳二元酸的回收率为 98.7% ，蛋白和菌体的去除率分别为 82.9% 和 100% 。
配制浓度为 57g /L 的十二碳二元酸发酵模拟液。取 5.5g 模拟液，加入 2.0g 磷酸氢二钾和 2.5g 正丁醇，搅拌均匀，室温静置，形成三相，上相为醇相，中相为富含十二碳二元酸相，下相为富含其它成分的盐相。十二碳二元酸的回收率为 88.6% 。

Claims (1)

1. 一种盐析萃取发酵液中有机酸的方法 ，其特征是： 发酵液是发酵模拟液、 经过滤除去菌体和蛋白的清液 或者是未经处理的发酵原液， 发酵液中有机酸浓度 为 20~170g/L ， pH 值为 2~10 。 向发酵液中添加 一种或二种以上 可溶性无机盐或有 机盐，以及单一或复合萃取剂， 搅拌均匀后于室温下静置分相 1~8h ，其中 盐质量百分数为 15~35% ，萃取剂的质量百分数为 15~35% ；当盐析萃取形成两相时，上相即为富含有机酸的溶剂相或萃取相，下相为富盐相或萃余相， 菌体和蛋白在两相之间形成固相层 ；当形成三相时，上相为溶剂相，中相为富含有机酸相，下相为富含其他成分的盐相。
2. 根据权利要求 1 所述的 一种盐析萃取发酵液中有机酸的方法 ，其特征是可溶性无机盐为 氯化钠、硫酸铵、硫酸钠、碳酸钠、碳酸钾、磷酸钠、磷酸钾、磷酸氢二钠、磷酸氢二钾、磷酸二氢钠或磷酸二氢钾等。可溶性有机盐为醋酸钠、醋酸钾、柠檬酸钠、柠蒙酸钾、草酸钠或草酸钾。
3. 根据权利要求 1 或 2 所述的 一种盐析萃取发酵液中有机酸的方法 ，其特征是萃取剂为甲醇、乙醇、正丙醇、异丙醇、正丁醇、异丁醇、丙酮、乙二醇、乙醚、乙酸甲酯或乙酸乙酯。
4. 根据权利要求 1 所述的 一种盐析萃取发酵液中有机酸的方法 ，其特征是 盐析 萃取可采取间歇萃取、连续萃取或多级萃取。
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