CN103601647B - Method for desalting and decolorizing L-alanine fermentation liquor produced by genetically engineered bacteria - Google Patents

Method for desalting and decolorizing L-alanine fermentation liquor produced by genetically engineered bacteria Download PDF

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CN103601647B
CN103601647B CN 201310611576 CN201310611576A CN103601647B CN 103601647 B CN103601647 B CN 103601647B CN 201310611576 CN201310611576 CN 201310611576 CN 201310611576 A CN201310611576 A CN 201310611576A CN 103601647 B CN103601647 B CN 103601647B
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alanine
dlol
desalted
decoloration
resin
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CN103601647A (en )
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洪厚胜
闵兆升
郭会明
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南京工业大学
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Abstract

本发明公开了一种基因工程菌产L-丙氨酸发酵液脱盐脱色的方法,该方法简单方便,可节约下游分离纯化的成本。 The present invention discloses a method of producing genetically engineered bacteria fermentation desalted decoloration L- alanine, which method is simple, cost savings downstream separation and purification. 本发明的基因工程菌产L-丙氨酸发酵液脱盐脱色的方法,其主要是使用D101大孔树脂对基因工程菌产L-丙氨酸发酵液进行脱盐脱色处理。 The method of the present invention genetically engineered bacteria producing L- Alanine desalted decoloration fermentation liquid, which is primarily genetically engineered bacteria producing L- phenylalanine fermentation broth was desalted decoloration treatment with D101 macroporous resin.

Description

一种基因工程菌产L-丙氨酸发酵液脱盐脱色的方法 An engineered L- phenylalanine producing bacterium fermentation method desalted decoloration

技术领域 FIELD

[0001] 本发明涉及一种发酵液脱盐脱色的方法,更具体地说涉及一种基因工程菌产L-丙氨酸发酵液脱盐脱色的方法。 [0001] The present invention relates to a method of decolorizing the fermentation broth desalting, and more particularly relates to a method for producing genetically engineered bacteria L- Alanine desalted decoloration of fermentation broth.

背景技术 Background technique

[0002] L-丙氨酸又称α -氨基丙酸,分子式为C3H7NO2,易溶于水,微溶于乙醇,不溶于乙醚或丙酮,含有较高的甜味,具有旋光性,是组成蛋白质的20种L型α氨基酸之一。 [0002] L- alanine, also known as α - amino acid, the formula of C3H7NO2, soluble in water, slightly soluble in ethanol, insoluble in ether or acetone, containing a high sweetness, optically active, is composed of a protein one of the amino acid L-α 20 species. L-丙氨酸虽然为人体非必需氨基酸,但却为人体血液中含量最高氨基酸,有重要的生理作用。 L- alanine Although the human body non-essential amino acids, but the body is the highest blood levels of amino acids, important physiological role. L-丙氨酸在生物体内转氨反应中为最重要的氨基供体,也可作为血液中氮的优良运输工具。 L- Alanine aminotransferase reaction in vivo, the most important amino donor, but also as an excellent vehicle nitrogen in the blood. 与糖代谢有密切关联,在葡萄糖-丙氨酸循环中,保持低血氨水平,同时还有产糖作用。 It is closely associated with glucose metabolism, glucose - alanine cycle, maintain a low blood ammonia levels, as well as sugar-producing role. L-丙氨酸在食品与医药行业有着极广的应用。 L- phenylalanine in the food and pharmaceutical industry has a very broad application. 在食品工业中,L-丙氨酸可用于食品饮料的防腐剂、风味调味料,加入少量的丙氨酸便可明显提高食品及饮料中的蛋白质利用率以及食品的营养价值,并且由于丙氨酸具有能够被生物体直接消化吸收的特点,饮用后能迅速恢复疲劳,振奋精神;L-丙氨酸具有较强的甜度,可作为食品甜味剂使用,在复配甜味剂中加入少量L-丙氨酸能使得甜度提高、甜味柔和如同天然甜昧剂,是合成高甜度阿力甜原料之一;L-丙氨酸还可改善后味,作酸味矫正剂,改善有机酸的酸味,使其更接近天然物质的酸味;L-丙氨酸还用于营养增补剂,制备氨基酸功能性保健酒等;L-丙氨酸在医药行业,广泛用于在临床输液,为患者提供能量和营养;L-丙氨酸还是生产维生素B6、合成泛酸钙及其它有机化合物的重要原料。 In the food industry, L- alanine can be used for food and beverage preservatives, flavoring, add a small amount of alanine can significantly improve the utilization of protein and nutritional value of food in the food and beverage, and because alanyl acid has to be digested and absorbed directly biometric characteristics, fatigue recovery after drinking quickly, vigor; L-alanine has strong sweetness, can be used as a food sweetener, the sweetener is added in compound L- alanine enables a small amount of sweetness enhanced sweetener soft as natural sweet ignorance agent, it is one of high intensity synthetic material alitame; L- alanine may further improve the taste, sour taste corrigent as improved sour organic acids, closer to natural substances acidity; L-alanine is further configured to nutritional supplements prepared amino functional health wine; L-alanine in the pharmaceutical industry, widely used in clinical infusion, provide energy and nutrients for the patient; L-alanine is an important raw material of vitamin B6, and other synthetic organic compounds of calcium pantothenate production.

[0003]目前,国内L-丙氨酸的生产绝大多数是采用生物发酵的方式。 [0003] Currently, the vast majority of domestic production of L- phenylalanine is the use of biological fermentation. 其除了对优良菌种选育、发酵工艺改进这些上游技术的探索,L-丙氨酸的下游分离提纯方式也是生物发酵生产L-丙氨酸的重要研宄内容和关键技术之一。 In addition to its excellent strain selection, fermentation processes these upstream explore improved technologies, L- Alanine downstream separation and purification of produced L- Alanine is one embodiment of an important study based on the content key and fermentation techniques. 微生物发酵液中存在大量色素分子和高盐成分,色素乃是本身有颜色并能使其它物质着色的高分子有机化合物。 Microbial broth dye molecules and a large number of high salt content, and the color of the dye itself but other substances can coloring high molecular organic compound. 它是在发酵过程中所产生的代谢产物,与菌种和发酵条件有关。 It is a metabolic product generated by the fermentation process, the bacteria and the fermentation conditions. 在发酵液预处理及提炼过程中,大部分的杂质及色素已被去除,但仍有少量色素存在,如果在提炼中使用质量较差的原材料,也会带进一些色素。 In the fermentation broth pretreatment and refining process, most of the impurities and pigments have been removed, but there is still a small amount of pigment, if you use poor quality raw material in the refining, also brought some of the pigment. 各种色素中生色基团和助基团一般都含有数量较多的各种极性基团(如-C00H、-NH2, -OH等)。 Chromophore group various pigments and co-groups generally contain a larger number of various polar groups (e.g. -C00H, -NH2, -OH, etc.). 如若不能有效去除这些色素分子和高盐成分,就会增加后续下游过程中产品的分离、提纯、结晶的难度,进而影响产品的纯度和品质。 Should not effectively remove the high salt content and the dye molecules, it will increase the separation of the product in subsequent downstream processes, purification, crystallization difficult, and thus affect the purity and quality of the product. L-丙氨酸的生产成本主要由上游发酵过程和下游分离提纯过程费用组成,其中,下游过程所占费用的比例高达60%〜70%。 L- Alanine production costs mainly by the upstream and downstream separation and purification fermentation process cost components, wherein the proportion of the cost of downstream processes share up to 60% ~ 70%. 通常L-丙氨酸脱色普遍采用的脱色方式是活性炭脱色。 Bleaching decolorization manner generally L- Alanine is commonly used active carbon. 这是因为活性炭比表面积大,吸附力强。 This is because the large surface area of ​​activated carbon adsorption and strong. 但是其也存在着不足与问题,其对溶液的黏度有一定的要求,黏度较高时需要使样液体升温到一定的温度才利于吸附和过滤,另外活性炭不可回收利用,且容易吸附产品,造成产品损失较大。 But there are also shortcomings and problems, which have certain requirements on the solution viscosity, the liquid sample needs to be heated only to a temperature conducive to higher viscosity adsorption and filtration, activated carbon can not be recycled further, and easily absorbed product, resulting in greater product loss. 而且活性炭脱色方式不能有效减少母液中高盐成分。 And decolorization method can not effectively reduce the high salt content in the mother liquor. 因此如何在节约成本的基础上改进分离提纯的方式,提高成品的品质已成为微生物发酵生产丙氨酸的关键问题之一。 Therefore, how to improve the separation and purification of the way on the basis of cost, improve the quality of the finished product has become one of the key problems of microbial fermentation production of phenylalanine.

[0004] DlOl大孔吸附树脂则是苯乙烯型非极性共聚体,适用范围比较广谱,对于不带极性或弱极性的有机化合物,普遍吸附能力强,目前,广泛应用在医药行业,特别对皂甙、黄酮这类中草药的分离提纯效果尤佳,例如:人参皂甙、三七皂甙、薯蓣皂甙、银杏黄酮等。 [0004] DlOl macroreticular resin is a styrene copolymer nonpolar, the scope of a broad spectrum comparison, with no respect to the polar or weakly polar organic compounds, generally strong adsorption capacity, at present, widely used in the pharmaceutical industry , especially for the purification effect separation of saponins, flavonoids such a plus herbs such as: ginsenosides, notoginseng saponins, dioscin, flavonoids and the like. 但是,目前还没有关于将DlOl大孔吸附树脂用于类似L-丙氨酸发酵液这类生物制品的脱盐脱色的相关专利和报道。 However, there is no regard to DlOl macroporous resin for desalting reports related patents and decolorization of such biological products like L- Alanine fermentation broth.

发明内容 SUMMARY

[0005] 本发明的目的是解决上述现有技术中存在的问题与不足,提供一种基因工程菌产L-丙氨酸发酵液脱盐脱色的方法,该方法简单方便,可节约下游分离纯化的成本。 [0005] The object of the present invention is to solve the above-described prior art problems and shortcomings, there is provided a method for producing genetically engineered bacteria fermentation desalted decoloration L- alanine, which method is simple, can be isolated and purified savings downstream cost.

[0006] 本发明是通过以下技术方案实现的: [0006] The present invention is achieved by the following technical solutions:

[0007] 本发明的基因工程菌产L-丙氨酸发酵液脱盐脱色的方法,其主要是使用DlOl大孔树脂对基因工程菌产L-丙氨酸发酵液进行脱盐脱色处理。 [0007] The present invention genetic engineering methods L- alanine fermentation bacteria producing desalted decoloration, which is mainly used for macroporous resin DlOl genetic engineering bacteria producing L- Alanine fermentation broth was desalted decoloration process.

[0008] 本发明的基因工程菌产L-丙氨酸发酵液脱盐脱色的方法,其进一步的方案是所述的使用DlOl大孔树脂对基因工程菌产L-丙氨酸发酵液进行脱盐脱色处理包括以下步骤: [0008] The present invention genetic engineering methods L- phenylalanine producing bacterium fermentation desalted decoloration, further solution is the use of macroporous resins DlOl L- phenylalanine producing genetically engineered bacteria broth was desalted decoloration process comprising the steps of:

[0009] 基因工程菌产L-丙氨酸发酵液预处理:将基因工程菌产L-丙氨酸发酵液在高速低温离心机下离心以去除包含菌体在内的杂质,收集得到上清液;将上清液中丙氨酸浓度调整至70〜80mg/mL,pH值调整至2.0〜3.0待用; [0009] genetically engineered bacteria producing L- phenylalanine fermentation liquid pretreatment: the genetically engineered bacteria producing L- Alanine low-speed centrifuge fermentation broth was centrifuged to remove impurities including the bacterial cells contain, the supernatant was collected liquid; alanine concentration in the supernatant was adjusted to 70~80mg / mL, pH value adjusted to 2.0~3.0 stand;

[0010] DlOl大孔树脂预处理:将DlOl大孔树脂装柱,并进行乙醇浸泡24h后水洗至流出液澄清后,再经过4%HC1流洗、水洗至中性、4%NaOH流洗、水洗至中性后的过程得到预处理DlOl大孔树脂待用; [0010] DlOl macroporous resin Pretreatment: DlOl macroporous resin packed in a column, and washed with ethanol and soaked for 24h to effluent clarification, then after the elution 4% HC1, washed with water until neutral, 4% NaOH wash stream, process after water until neutral to give pretreated macroporous resin DlOl stand;

[0011] 脱盐脱色处理:将基因工程菌产L-丙氨酸发酵液预处理步骤中已处理好的L-丙氨酸发酵液以流速为0.5〜2.5mL/min,温度为35〜65°C条件下进入经过预处理的DlOl大孔树脂进行脱盐脱色,色素有机大分子及含高盐类杂质吸附在大孔树脂上,收集分离后的L-丙氨酸发酵液。 [0011] Desalting bleaching process: the genetically engineered bacteria producing L- Alanine broth pretreatment step already processed L- phenylalanine fermentation liquid at a flow rate 0.5~2.5mL / min, a temperature of 35~65 ° C under the conditions for entering desalted decoloration, organic molecules and the dye-containing high salt impurities adsorbed on a macroporous resin was collected through the separated fermentation broth L- alanine DlOl pretreated macroporous resin.

[0012] 本发明的基因工程菌产L-丙氨酸发酵液脱盐脱色的方法,其更进一步的技术方案是所述的高速低温离心机的转速为10000〜12000r/min,离心时间为5min。 The method of [0012] genetically engineered bacteria of the present invention, the fermentation yield L- Alanine desalted decoloration, which is a further aspect of the low temperature high speed centrifuge speed is 10000~12000r / min, centrifugation time was 5min.

[0013] 本发明的基因工程菌产L-丙氨酸发酵液脱盐脱色的方法,其更进一步的技术方案还可以是所述的DlOl大孔树脂预处理时在装柱前先于柱中装入1/3〜1/4体积的水,然后加入DlOl树脂,树脂装柱量为树脂膨胀后柱体积的80%。 [0013] When the present invention genetic engineering methods L- phenylalanine producing bacterium fermentation desalted decoloration, which further aspect of the DlOl may also be pretreated prior to macroporous resin packed columns are installed prior to into 1 / 3~1 / 4 volumes of water, then added DlOl resin, amount of the resin packed column volumes of 80% expansion of the resin.

[0014] 本发明的基因工程菌产L-丙氨酸发酵液脱盐脱色的方法,其更进一步的技术方案是脱盐脱色处理过程中,在DlOl大孔树脂吸附饱和后,用50〜99%乙醇、0.5〜2.0mL/min的流速进行树脂的洗脱使DlOl大孔树脂再生。 [0014] The present invention genetic engineering methods L- phenylalanine producing bacterium fermentation desalted decoloration, which further aspect desalination bleaching process, after DlOl macroporous resin saturated with 50~99% ethanol , 0.5~2.0mL / min flow rate so that the resin elution was carried out DlOl macroporous resin regeneration.

[0015] 与现有技术相比本发明具有以下有益效果: [0015] has the following advantages compared with the prior art:

[0016] 利用DlOl大孔吸附树脂进行脱盐脱色,所吸附的色素和杂质容易洗脱,80%乙醇洗脱即可,酸碱重复处理可实现树脂的再生。 [0016] DlOl using macroporous adsorption resin desalted decoloration, and impurities adsorbed dye easy elution to 80% ethanol elution, repetitive processing of the recycled resin acid can be achieved. DlOl大孔树脂对L-丙氨酸吸附量极少甚至没有吸附,产品保留率达到94%以上,不会造成产品损失。 DlOl macroporous adsorption resin L- phenylalanine little or no adsorption, product retention rate of more than 94%, without loss of product. DlOl大孔树脂较其它普通树脂在进行发酵液脱色的同时,还能脱盐,降低发酵液电导率。 DlOl macroporous resin during fermentation decoloration than other common resins, while also desalted broth reduced conductivity. DlOl大孔树脂的再生率较高,达到95%以上,可以重复使用数十次。 Higher regeneration rate DlOl macroporous resin, more than 95%, can be reused dozens of times. 使用DlOl大孔树脂对L-丙氨酸进行脱盐脱色,效果优于活性炭脱色,可节约下游分离纯化的成本,适合工业化规模生产用。 Use of macroporous resin DlOl L- Alanine desalted decoloration, better than decolorization, separation and purification can be cost downstream, for industrial scale production.

附图说明 BRIEF DESCRIPTION

[0017] 图1为本发明实施例1中DlOl大孔树脂对L-丙氨酸脱色的脱色率随温度变化示意图。 [0017] FIG 1 decolorization Example 1 DlOl macroporous resin decolorization of L- Alanine schematic embodiment of the present invention varies with temperature.

[0018] 图2为本发明实施例2中DlOl大孔树脂对L-丙氨酸脱色的脱色率随上样流速变化示意图。 [0018] FIG. 2 decolorization embodiment DlOl macroporous two pairs of L- Alanine decolorized loading flow rate variation with the embodiment of the present invention, a schematic diagram.

[0019] 图3为本发明实施例3中DlOl大孔树脂对L-丙氨酸脱色的解析率随乙醇浓度变化示意图。 [0019] FIG. 3 desorption ratio in Example 3 macroporous resin DlOl L- Alanine decolorized with alcohol concentration schematic embodiment of the present invention.

具体实施方式 detailed description

[0020] 在实施例中DlOl大孔吸附树脂均为市售产品。 [0020] In an embodiment DlOl macroporous resin are commercially available products.

[0021] 实施例1 [0021] Example 1

[0022] (I)将L-丙氨酸发酵液进行预处理,在高速低温离心机下离心以去除菌体等杂质,收集上清液; [0022] (I) The fermentation broth pretreated L- Alanine, in low-speed centrifuge was centrifuged to remove cells and other impurities, the supernatant was collected;

[0023] (2)将上述上清液中L-丙氨酸浓度调整至75mg/mL,pH值调整至3.0 ; [0023] (2) The above supernatant L- alanine concentration was adjusted to 75mg / mL, pH value adjusted to 3.0;

[0024] (3)将DlOl大孔树脂装柱,并进行乙醇浸泡24h后水洗至流出液澄清后,依次用4%HC1流洗,水洗至中性,4%NaOH流洗,水洗至中性的预处理过程; [0024] (3) The DlOl macroporous resin packed in a column, and washed with ethanol to 24h after soaking clarified effluent, flows sequentially washed with 4% HC1, washed with water until neutral, 4% NaOH wash stream, washed with water until neutral pretreatment process;

[0025] (4)控制处理后的发酵液流速为1.5mL/min,温度分别为35、40、45、50、55、60°C、65°C下,进入DlOl大孔树脂进行脱盐脱色,得丙氨酸发酵液,计算脱色率。 [0025] (4) controls the flow rate of the fermentation broth after the treatment was 1.5mL / min, respectively, the temperature of 35,40,45,50,55,60 ° C, at 65 ° C, enters the macroporous resin DlOl desalted decoloration, phenylalanine fermentation broth obtained, decolorizing rate is calculated. 图1是DlOl大孔树脂动态脱色试验得到的温度对脱色率变化曲线图,可以看出,温度对DlOl大孔树脂的吸附性能的影响比较大,温度在50°C左右脱色性能卓越。 FIG 1 is a macroporous resin DlOl dynamic crock obtained temperature variation graph showing the decolorization rate can be seen, the effect of temperature on the adsorption properties DlOl the macroporous resin is relatively large, a temperature of about 50 ° C excellent bleaching performance.

[0026] (5)树脂吸附饱和后,用80%乙醇、1.0mL/min的流速进行树脂的洗脱。 [0026] (5) Adsorption saturated with 80% ethanol, 1.0mL / min of flow rate of the resin was eluted.

[0027] 上述方法中,离心机转速为11000r/min,离心时间为5min。 [0027] In the above method, the centrifuge speed is 11000r / min, centrifugation time was 5min.

[0028] 实施例2 [0028] Example 2

[0029] (I)将L-丙氨酸发酵液进行预处理,在高速低温离心机下,以12000r/min离心5min,以去除菌体等杂质,收集上清液; [0029] (I) The fermentation broth pretreated L- Alanine, at a low temperature high speed centrifuge to / centrifuged at 12000r min 5min, to remove impurities such as fungi, the supernatant was collected;

[0030] (2)将上述上清液中L-丙氨酸浓度调整至75mg/mL,pH值调整至3.0 ; [0030] (2) The above supernatant L- alanine concentration was adjusted to 75mg / mL, pH value adjusted to 3.0;

[0031] (3)将DlOl大孔树脂装柱,并进行乙醇浸泡24h后水洗至流出液澄清后,依次用4%HC1流洗,水洗至中性,4%NaOH流洗,水洗至中性的预处理过程; [0031] (3) The DlOl macroporous resin packed in a column, and washed with ethanol to 24h after soaking clarified effluent, flows sequentially washed with 4% HC1, washed with water until neutral, 4% NaOH wash stream, washed with water until neutral pretreatment process;

[0032] (4)控制温度在30°C 下,以0.5mL/min、lmL/min、1.5mL/min、2mL/min、2.5mL/min的流速进入DlOl大孔树脂进行脱盐脱色,得丙氨酸发酵液,计算脱色率。 [0032] (4) controlling the temperature at 30 ° C, at 0.5mL / min, lmL / min, 1.5mL / min, a flow rate of 2mL / min, 2.5mL / min into the macroporous resin DlOl desalted decoloration give propyl acid fermentation broth, decolorizing rate is calculated. 图2是DlOl大孔树脂动态脱色试验得到的上样流速对脱色率变化曲线图,可以看出,流速在1.5mL/min左右脱色性能卓越。 FIG 2 is a macroporous resin DlOl crock obtained dynamic loading flow rate on the decolorization rate variation graph, it can be seen, excellent bleaching performance at a flow rate of about 1.5mL / min.

[0033] (5)树脂吸附饱和后,用80%乙醇、1.0mL/min的流速进行树脂的洗脱。 [0033] (5) Adsorption saturated with 80% ethanol, 1.0mL / min of flow rate of the resin was eluted.

[0034] 上述方法中,离心机转速为12000r/min,离心时间为5min。 [0034] The above-described method, the centrifuge speed is 12000r / min, centrifugation time was 5min.

[0035] 实施例3 [0035] Example 3

[0036] (I)将L-丙氨酸发酵液进行预处理,在高速低温离心机下,以12000r/min离心5min,以去除菌体等杂质,收集上清液; [0036] (I) The fermentation broth pretreated L- Alanine, at a low temperature high speed centrifuge to / centrifuged at 12000r min 5min, to remove impurities such as fungi, the supernatant was collected;

[0037] (2)将上述上清液中L-丙氨酸浓度调整至75mg/mL,pH值调整至3.0 ; [0037] (2) The above supernatant L- alanine concentration was adjusted to 75mg / mL, pH value adjusted to 3.0;

[0038] (3)将DlOl大孔树脂装柱,并进行乙醇浸泡24h后水洗至流出液澄清后,依次用4%HC1流洗,水洗至中性,4%NaOH流洗,水洗至中性的预处理过程; [0038] (3) The DlOl macroporous resin packed in a column, and washed with ethanol to 24h after soaking clarified effluent, flows sequentially washed with 4% HC1, washed with water until neutral, 4% NaOH wash stream, washed with water until neutral pretreatment process;

[0039] (4)控制温度在30°C下,以1.5mL/min的流速进入DlOl大孔树脂进行脱盐脱色,得丙氨酸发酵液; [0039] (4) controlling the temperature at 30 ° C, at a flow rate of 1.5mL / min into the macroporous resin DlOl desalted decoloration give alanine fermentation broth;

[0040] (5)树脂吸附饱和后,用浓度分别为50%、60%、70%、80%、99%乙醇、1.0mL/min的流速进行树脂的洗脱,计算解析率。 [0040] (5) saturated adsorption resin, at a concentration of 50%, 60%, 70%, 80%, 99% ethanol, flow rate, 1.0mL / min eluted the resin, desorption ratio is calculated. 图3是DlOl大孔树脂动态脱色试验得到的乙醇浓度对解析率变化曲线图,可以看出,乙醇浓度在80%左右解析性能卓越。 FIG 3 is a macroporous resin ethanol concentration DlOl dynamic crock obtained for parsing rate profiles can be seen, analytical high performance ethanol concentration of about 80%.

[0041] 上述方法中,离心机转速为10000r/min,离心时间为5min。 [0041] The above-described method, the centrifuge speed is 10000r / min, centrifugation time was 5min.

Claims (4)

  1. 1.一种基因工程菌产L-丙氨酸发酵液脱盐脱色的方法,其特征在于使用DlOl大孔树脂对基因工程菌产L-丙氨酸发酵液进行脱盐脱色处理;所述的使用DlOl大孔树脂对基因工程菌产L-丙氨酸发酵液进行脱盐脱色处理包括以下步骤: 基因工程菌产L-丙氨酸发酵液预处理:将基因工程菌产L-丙氨酸发酵液在高速低温离心机下离心以去除包含菌体在内的杂质,收集得到上清液;将上清液中丙氨酸浓度调整至70〜80mg/mL,pH值调整至2.0〜3.0待用; DlOl大孔树脂预处理:将DlOl大孔树脂装柱,并进行乙醇浸泡24h后水洗至流出液澄清后,再经过4% HCl流洗、水洗至中性、4% NaOH流洗、水洗至中性后的过程得到预处理DlOl大孔树脂待用; 脱盐脱色处理:将基因工程菌产L-丙氨酸发酵液预处理步骤中已处理好的L-丙氨酸发酵液以流速为0.5〜2.5mL/min,温度为35〜65°C条件下进入经过预处理的DlOl Use according DlOl; CLAIMS 1. A method of genetically engineered bacteria producing L- phenylalanine fermentation liquid desalted decoloration, characterized in that a macroporous resin DlOl genetic engineering bacteria producing L- Alanine treated fermentation broth was desalted decoloration macroporous resin genetic engineering bacteria producing L- alanine fermentation broth was desalted decoloration process comprising the steps of: L- phenylalanine producing bacterium fermentation broth pretreated genetic engineering: the genetically engineered bacteria producing L- alanine fermentation broth low-speed centrifuge was centrifuged to remove impurities including the bacterial cells contain, the supernatant was collected; supernatants adjusted to a concentration of alanine 70~80mg / mL, pH value adjusted to 2.0~3.0 stand; DlOl macroporous resin pretreatment: DlOl macroporous resin packed in a column, and washed with ethanol and soaked for 24h to effluent clarification, then flows through the wash 4% HCl, water until neutral, 4% NaOH wash stream, washed with water until neutral after the process has been pretreated macroporous resin DlOl stand; desalted decoloration process: the genetically engineered bacteria producing L- alanine broth pretreatment step already processed L- phenylalanine fermentation liquid at a flow rate of 0.5~2.5 mL / min, the temperature into the pretreated DlOl conditions at 35~65 ° C 孔树脂进行脱盐脱色,收集分离后的L-丙氨酸发酵液。 Amberlyst desalted decoloration collected alanine L- broth after separation.
  2. 2.根据权利要求1所述的基因工程菌产L-丙氨酸发酵液脱盐脱色的方法,其特征在于所述的高速低温离心机的转速为10000〜12000r/min,离心时间为5min。 2. The method of claim 1 genetically engineered bacteria producing L- Alanine fermentation desalted decoloration claim, wherein said low-speed centrifuge speed is 10000~12000r / min, centrifugation time was 5min.
  3. 3.根据权利要求1所述的基因工程菌产L-丙氨酸发酵液脱盐脱色的方法,其特征在于所述的DlOl大孔树脂预处理时在装柱前先于柱中装入1/3〜1/4体积的水,然后加入DlOl树脂,树脂装柱量为树脂膨胀后柱体积的80%。 3. The method of producing L- Alanine bacteria fermentation desalted decoloration of genetic engineering according to claim 1, characterized in that the initially charged in a column packed before DlOl when pretreated macroporous resin according to 1 / 3~1 / 4 volumes of water, then added DlOl resin, amount of the resin packed column volumes of 80% expansion of the resin.
  4. 4.根据权利要求1所述的基因工程菌产L-丙氨酸发酵液脱盐脱色的方法,其特征在于脱盐脱色处理过程中,在DlOl大孔树脂吸附饱和后,用50〜99%乙醇、0.5〜2.0mL/min的流速进行树脂的洗脱使DlOl大孔树脂再生。 According to claim 1, said genetic engineering method for producing L- Alanine bacteria fermentation broth is desalted decoloration, desalting wherein bleaching process, after DlOl macroporous resin saturated with 50~99% ethanol, 0.5~2.0mL / min flow rate so that the resin elution was carried out DlOl macroporous resin regeneration.
CN 201310611576 2013-11-27 2013-11-27 Method for desalting and decolorizing L-alanine fermentation liquor produced by genetically engineered bacteria CN103601647B (en)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1099268A (en) * 1994-03-30 1995-03-01 三门峡市化工研究院 Process for production of amino acid oral liquid
CN101275149A (en) * 2008-05-08 2008-10-01 江南大学 Preparation for asparaginic acid protease inhibitors
CN103387502A (en) * 2012-05-09 2013-11-13 上海医药工业研究院 Method for extracting L-alanine from L-alanine fermentation liquid

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CA2753004A1 (en) * 2009-02-25 2010-09-02 Danisco A/S Separation process

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1099268A (en) * 1994-03-30 1995-03-01 三门峡市化工研究院 Process for production of amino acid oral liquid
CN101275149A (en) * 2008-05-08 2008-10-01 江南大学 Preparation for asparaginic acid protease inhibitors
CN103387502A (en) * 2012-05-09 2013-11-13 上海医药工业研究院 Method for extracting L-alanine from L-alanine fermentation liquid

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