WO2008134936A1 - Procédé d'extraction de thréonine à partir de liqueur de fermentation de thréonine - Google Patents

Procédé d'extraction de thréonine à partir de liqueur de fermentation de thréonine Download PDF

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Publication number
WO2008134936A1
WO2008134936A1 PCT/CN2008/000874 CN2008000874W WO2008134936A1 WO 2008134936 A1 WO2008134936 A1 WO 2008134936A1 CN 2008000874 W CN2008000874 W CN 2008000874W WO 2008134936 A1 WO2008134936 A1 WO 2008134936A1
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WIPO (PCT)
Prior art keywords
threonine
temperature
crystallization
fermentation broth
crystal
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PCT/CN2008/000874
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English (en)
Chinese (zh)
Inventor
Dehui Wang
Dongshu Jia
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Changchun Dacheng Industrial Group Company Limited
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Publication of WO2008134936A1 publication Critical patent/WO2008134936A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P13/00—Preparation of nitrogen-containing organic compounds
    • C12P13/04—Alpha- or beta- amino acids
    • C12P13/08—Lysine; Diaminopimelic acid; Threonine; Valine
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C227/00—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C227/38—Separation; Purification; Stabilisation; Use of additives
    • C07C227/40—Separation; Purification
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C227/00—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C227/38—Separation; Purification; Stabilisation; Use of additives
    • C07C227/40—Separation; Purification
    • C07C227/42—Crystallisation

Definitions

  • the present invention relates to a method for fermenting threonine by threonine fermentation, whereby a threonine crystal having a high content of threonine can be obtained, and the threonine crystal can be used as an animal feed additive.
  • Threonine is one of the essential amino acids in humans and animals, but threonine cannot be synthesized in the body and must be taken from food. Therefore, in recent years, threonine has been widely used as an additive in the fields of food, health care, and feed industries. In the animal term, threonine is the second limiting amino acid. In recent years, as the word additive, as the amount of lysine has increased year by year, the amount of threonine has increased year by year. China's threonine feed additive mainly relies on imports. At present, medical threonine is mostly chemically synthesized. The production of a large amount of threonine used as a food or feed additive is mainly produced by fermentation.
  • the fermentation method Compared with the chemical synthesis method, the fermentation method has a large production scale, high yield and low cost, which can meet the increasing demand.
  • the process for producing threonine by fermentation is not mature enough, and various production steps such as filtration, concentration and crystallization are to be improved and optimized.
  • CN200610014324.4 discloses a method for separating and extracting L-threonine from a fermentation liquid, wherein the fermentation liquid is filtered by a metal membrane to improve the filtration process and improve the production efficiency, and the purity of the threonine crystal product can reach 99. %the above.
  • the inventors have made a lot of research and experiments on the process of extracting the threonine crystal product from the fermentation liquid, and have successfully completed the industrial scale production experiment, the product quality is good, the yield is high, and there is no Environmental pollution. Summary of the invention
  • the step (1) further comprises: after subjecting the threonine fermentation broth to high temperature sterilization, adjusting the pH to 4.0 to 5.0, preferably about 4.5, with sulfuric acid, and then removing the cells by filtration through an inorganic ceramic membrane.
  • the membrane can retain a protein having a molecular weight of 300 kD, and a threonine fermentation liquid having a light transmittance of 87% or more is obtained.
  • the bacterial protein removed by membrane filtration can be made into a fertilizer after being filtered by a three-stage membrane.
  • the inorganic ceramic membrane is obtained by sintering an oxide such as alumina, titania or zirconia at a high temperature, and is a porous ceramic filter material having a porous structure in which the porous support layer, the filter layer and the microporous membrane layer are asymmetrically distributed. Filtration accuracy can range from microfiltration to ultrafiltration to sodium filtration. Ceramic membrane filtration is a fluid separation process in the form of "cross-flow filtration" in which a raw material liquid flows at a high speed in a membrane tube, and a clarified permeate containing a small molecular component is driven through the membrane in a direction perpendicular thereto.
  • the turbid concentrate containing the macromolecular component is trapped by the membrane, thereby allowing the fluid to be separated, concentrated and purified.
  • inorganic ceramic membranes have many advantages, such as excellent chemical stability, high temperature resistance, high mechanical strength, narrow pore size distribution, high separation precision, easy cleaning, etc., compared with other filter materials, inorganic ceramics.
  • the membrane is separated, the obtained separation product is high in purity and easier to concentrate, so it is widely used in pharmaceutical, bioengineering, electronics and other industries.
  • the inorganic ceramic membrane is used for the filtration operation, the operating temperature and pressure can be carried out according to the instructions of the product specification or the conventional operating conditions.
  • the membrane filtration temperature is about 75 ° C
  • the pressure inlet pressure is about 0.7 mpa/m. 2
  • the filming pressure is about 0.4mpa/m 2 .
  • the activated carbon used in the decolorization step of the activated carbon in the step (2) may be a conventionally used activated carbon, preferably two activated carbons having different particle sizes are used together, and more preferably the particle size is 200 ⁇ 300 mesh and 18 mesh activated carbon are used together. Among them, 200 ⁇ 300 mesh activated carbon can be used to remove about 80% of the color. With 18 mesh activated carbon, about 15% of the color can be removed. Decolorization with activated carbon can greatly improve the decolorization efficiency of activated carbon.
  • the concentration under reduced pressure according to the step (3) can be carried out by a conventional concentration method, preferably by using a multi-effect falling film evaporation apparatus and method, for example, a four-effect falling film evaporator for concentration, and a four-effect falling film.
  • the temperatures of the evaporation system are approximately: one-effect evaporation temperature of 90 ° C, two-effect evaporation temperature of 60 ⁇ , three-effect evaporation temperature of 80 ° C, and four-effect evaporation temperature of 70 ° C.
  • concentration of the obtained concentrated liquid reaches 25 Bo or more, preferably 25 to 26 Bo, the concentration step can be completed.
  • the temperature-lowering crystallization described in the step (4) is carried out by using a gradient cooling crystallization method in the crystallization tank, and the gradient cooling crystallization condition is an initial temperature of 70 ° C and a cooling rate of 2.5-2.7 ° C / h. , the minimum temperature is 5-10 ° C, maintained for 15-30 hours, while stirring, the small grains are further grown to facilitate separation.
  • the preferred conditions for the gradient cooling crystallization are: an initial temperature of 70 ° C, a cooling rate of 2.57 ° C / h, a minimum temperature of 9 ° C, a temperature of 20 h, and a stirring speed of about 30 rpm.
  • an initial temperature of 70 ° C a cooling rate of 2.57 ° C / h
  • a minimum temperature of 9 ° C a temperature of 20 h
  • a stirring speed of about 30 rpm.
  • the purity of the resulting threonine crystal can reach 98.5% or more, and the threonine extraction rate can reach 95% or more.
  • the method for extracting threonine crystals from the threonine fermentation broth of the present invention comprises the following steps:
  • the pH is adjusted to 4.5 with sulfuric acid, and then the inorganic protein membrane is used to filter and remove impurities such as bacterial proteins, and the membrane can retain a protein having a molecular weight of 300 kD to obtain a light transmittance. More than 87% of threonine fermentation broth;
  • the decolorized fermentation broth enters the four-effect falling film evaporator, so that the concentration of the fermentation liquid reaches 25 ⁇ 26Bo°; (4) The above fermentation broth is passed into a gradient cooling crystallization tank for temperature crystallization, and the gradient crystallization conditions are: starting temperature 70 ° C, cooling rate is 2.5-2.7 ° C / h, minimum temperature 5 - 10 ° C, Maintain for 15-30 hours while stirring;
  • the primary mother liquor recovered in the step (5) can be recovered and reused, and the reuse method comprises the following steps: First, fermenting the sterilized and filtered threonine The clear liquid is combined and injected into the program of the method of the present invention for recycling; the second is to directly decolorize, concentrate, gradient crystallize, and centrifuge to obtain threonine crystal and secondary mother liquid, and the obtained crystal is obtained.
  • the finished threonine crystals are obtained by redissolving, crystallizing and separating and drying the crystals with water.
  • the method described in the latter is basically the same as the method for treating the fermentation liquid concentrate described above, that is, another independent process system is provided, and the operating conditions of each step are substantially the same as those described above.
  • the method for obtaining threonine by recrystallization from a mother liquor in the latter method is: decolorizing, concentrating, gradient cooling, and centrifuging the primary mother liquor to obtain threonine crystals and a second mother liquor.
  • the threonine content of the obtained threonine crystal is about 60%; the crystal is redissolved with water, that is, the crystal is dissolved by steam at 70-80 ° C with high-temperature hot water, for example, the obtained crystal is 2 tons.
  • the ratio of lm 3 water is dissolved in high temperature hot water at 70-80 ° C with steam, and then cooled again by gradient cooling, separation and drying to obtain the finished threonine crystal.
  • the primary mother liquor obtained in the step (5) is subjected to decolorization, concentration, gradient cooling crystallization and centrifugation to obtain a threonine product, and a secondary mother liquor is obtained in the above centrifugation process, and the secondary mother liquor is also obtained.
  • Recyclable for example, can be used to make fertilizer.
  • the secondary condensate of the concentration step in the method of the present invention can be fully utilized, and can be used as condensed water or as washing water in a post-treatment operation of threonine crystals.
  • the method of the invention has high yield and the threonine extraction rate can reach more than 95%; the yield, that is, the ratio of the total amount of the finished crystal product to the total amount of threonine in the clear liquid can reach above 91%; the product quality is good, the obtained product
  • the purity of threonine crystals is above 98.5 %; the crystal product particles are moderate, which is advantageous for separation, and is also advantageous for mixing with other ingredients in feed processing;
  • the comprehensive utilization of various by-products greatly reduces the production cost.
  • the primary mother liquor, the secondary mother liquor, and the bacterial protein dosage form in the production process can be comprehensively utilized as needed, and the threonine is recovered or made into a secondary product.
  • Grade products such as fertilizers
  • the secondary condensate obtained in the concentration step can be used as condensed water or washing water in the post-treatment of the threonine crystallization process.
  • the method of the invention is simple, easy to popularize, and can meet the increasing demand, and the production method of the invention is a typical environment-friendly production process, which is beneficial to environmental protection. - BRIEF DESCRIPTION OF THE DRAWINGS
  • Figure 1 is a flow chart showing the preparation of threonine crystals from a threonine fermentation broth according to the present invention.
  • Example 1 Preparation of threonine crystals from threonine fermentation broth
  • threonine fermentation broth Preparation of threonine fermentation broth according to a conventional method: using corn sugar as a main raw material, adding ammonium sulfate, ammonia water and supplemented with corn syrup as a nitrogen source, and adding a bacterium of the genus Brevibacterium to ferment a threonine fermentation liquid, the fermentation broth
  • the basic parameters are:
  • Threonine content 7 ⁇ 12g/100ml
  • the filtered broth take threonine broth 300m 3, wherein the threonine content of 9g / 100ml, pH value of 7.00, the 100 ° C high temperature sterilization, adjusted with food grade concentrated sulfuric acid to pH 4.5, followed by
  • the pump is pumped into a 300KD inorganic ceramic membrane (France, Novartis Orelis, model: K99BW) for filtration to remove impurities such as bacterial proteins.
  • the pumping pressure is 7kg, the pumping pressure is 4kg, and the temperature is 70°C. If the pressure is too high, the membrane will be broken and the filtration will be lost.
  • Decolorization of the fermentation broth The clarified fermentation broth is added with 200-300 mesh activated carbon at a temperature of 70 ° C, pH 4.5, clarified fermentation broth at a ratio of 1%, filtered, and then passed through two 18-mesh activated carbon.
  • the column is decolored, and the light transmittance of the decolorized fermentation broth is above 99%, which is favorable for forming large crystal grains.
  • the carbon column has a diameter of 3 m, a height of 5 m, and a pumping flow rate of 150 m 3 /h. 4.
  • Concentration of fermentation broth The decolorized fermentation broth enters the four-effect falling film evaporation system, the evaporation temperature of one effect is 90 ⁇ , the evaporation temperature of the second effect is 60°C, and the wave of the second-effect evaporation is between about 9 ⁇ 10.
  • a secondary condensate of 130 m 3 is obtained , which can be stored as condensed water or washing water for use.
  • Gradient cooling crystallization The crystal slurry obtained in the previous step is pumped from the second-effect evaporation pump into the gradient cooling crystallization tank, and the cooling water tube bundle of the crystallization tank is passed through the groundwater to cool down, the cooling rate is 2.570 ° C / h, and the stirring speed is 30 rpm.
  • the temperature is lower than 15 °C, due to the limitation of groundwater temperature, the ice water is used to cool down, and the cooling rate is constant.
  • the temperature reaches 9X:, the temperature is stirred for 20 hours to make the small grains grow further, so as to facilitate Separation.
  • Post-treatment of crystallization pump the crystal slurry into the piston to push the centrifuge to separate the crystal and the primary mother liquor, and wash it three times with the condensed water obtained in step 4 to wash away impurities such as inorganic salts and pigments attached to the crystal surface.
  • the threonine crystal is obtained after drying, and the crystal is dried and weighs 23.00 tons, and the content is 98.5% or more.
  • the centrifuge used for centrifugation of crystal is a self-unloading three-legged centrifuge with a separation factor of 0.5-0.6 and a screen mesh of 250 mesh. The screen is too small to dehydrate the crystal material, and it is easy to run the crystal material and waste. energy.
  • a secondary mother liquor is obtained when the crystal is separated, wherein the threonine content is 7.90 g/100 ml and the volume is 2.5 m 3 .
  • the above secondary mother liquor and the bacterial cells are mixed and granulated into a fertilizer.
  • the finished product of the threonine crystal obtained by the method of the present embodiment has a purity of 98.5% or more and has the following dry matter composition:

Abstract

La présente invention concerne un procédé d'extraction de la thréonine à partir d'une liqueur de fermentation de thréonine. Ce procédé comporte plusieurs opérations. On commence par filtrer au travers d'un film de céramique la liqueur de fermentation de thréonine stérilisée. On décolore la liqueur de fermentation de thréonine obtenir par un ajout de charbon actif puis on la verse dans un évaporateur à effets multiples de façon à la concentrer. On introduit ensuite la liqueur concentrée dans un cristallisoir à chute de température en gradient de façon à la cristalliser quand la concentration de la liqueur atteint les 25~26 sur l'échelle Baumé. Il est alors possible de séparer par centrifugation puis de sécher les cristaux de thréonine obtenus dont le degré de pureté atteint au minimum les 98,5%. La liqueur mère primaire obtenue à partir de l'étape de cristallisation peut être utilisée en cercle, et le produit cristallin de thréonine peut être obtenu par les procédés de décoloration, concentration, cristallisation par chute de température, centrifugation, re-résolution, séparation par centrifugation et séchage de ladite liqueur mère. Le procédé de l'invention donne un rendement de purification de la thréonine de plus de 95%, et résout les problèmes d'environnement causés par la décharge de liqueur mère en grandes quantités.
PCT/CN2008/000874 2007-04-29 2008-04-29 Procédé d'extraction de thréonine à partir de liqueur de fermentation de thréonine WO2008134936A1 (fr)

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CN200710097998.1 2007-04-29
CN200710097998A CN101293847B (zh) 2007-04-29 2007-04-29 由苏氨酸发酵液提取苏氨酸的方法

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CN106349095A (zh) * 2016-08-30 2017-01-25 呼伦贝尔东北阜丰生物科技有限公司 一种苏氨酸提取结晶工艺
CN112915563A (zh) * 2021-03-10 2021-06-08 郑州博大浓缩干燥设备有限公司 玉米浸泡液高效节能蒸发系统
CN113277955A (zh) * 2021-06-21 2021-08-20 通辽梅花生物科技有限公司 L-异亮氨酸的提取方法
CN113480444A (zh) * 2021-08-02 2021-10-08 齐齐哈尔龙江阜丰生物科技有限公司 一种节能减排的氨基酸发酵工艺
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CN110051634A (zh) * 2019-05-29 2019-07-26 绥化象屿金谷生化科技有限公司 一种l-苏氨酸湿法造粒方法
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CN106349095A (zh) * 2016-08-30 2017-01-25 呼伦贝尔东北阜丰生物科技有限公司 一种苏氨酸提取结晶工艺
CN106349095B (zh) * 2016-08-30 2018-01-23 呼伦贝尔东北阜丰生物科技有限公司 一种苏氨酸提取结晶工艺
CN112915563A (zh) * 2021-03-10 2021-06-08 郑州博大浓缩干燥设备有限公司 玉米浸泡液高效节能蒸发系统
CN113277955A (zh) * 2021-06-21 2021-08-20 通辽梅花生物科技有限公司 L-异亮氨酸的提取方法
CN113480444A (zh) * 2021-08-02 2021-10-08 齐齐哈尔龙江阜丰生物科技有限公司 一种节能减排的氨基酸发酵工艺
CN115029394A (zh) * 2022-05-12 2022-09-09 呼伦贝尔东北阜丰生物科技有限公司 苏氨酸母液分离提取工艺
CN115029400A (zh) * 2022-07-04 2022-09-09 安徽申亚农业发展有限公司 一种畜禽补铁用右旋糖酐铁的双发酵制备方法
CN117384095A (zh) * 2023-12-08 2024-01-12 山东联盟磷复肥有限公司 一种吡唑的脱色方法
CN117384095B (zh) * 2023-12-08 2024-03-12 山东联盟磷复肥有限公司 一种吡唑的脱色方法

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