阅读说明：本技术 一种从发酵液中提取维生素vb12的分离纯化方法 (Separation and purification method for extracting vitamin VB12 from fermentation liquor ) 是由 李俞胜 于 2021-08-05 设计创作，主要内容包括：本发明涉及生物发酵技术领域,尤其是一种从发酵液中提取维生素VB12的分离纯化方法,现提出如下方案,其包括将经过酸化后pH为2-5的第一发酵液导入预处理系统得到一级透析液；将一级透析液导入有机管式膜系统得到第一浓缩液和二级透析液；将二级透析液导入纳滤膜系统,除去二级透析液中的一价盐,得到第二浓缩液和三级透析液；将第二浓缩液导入到离子交换树脂系统得到四级透析液,对四级透析液萃取后结晶得到维生素VB12。本发明通过纳滤膜的道南效应去除一价盐,减轻后续的离子交换树脂系统的负荷,纳滤膜透析液浓缩后经过蒸发结晶得到氯化钠结晶盐,大大减轻了环保压力和淡水资源消耗量。(The invention relates to the technical field of biological fermentation, in particular to a separation and purification method for extracting vitamin VB12 from fermentation liquor, and provides the following scheme, which comprises the steps of introducing the first fermentation liquor with pH of 2-5 after acidification into a pretreatment system to obtain primary dialysate; introducing the primary dialysate into an organic tubular membrane system to obtain a first concentrated solution and a secondary dialysate; introducing the secondary dialysate into a nanofiltration membrane system, and removing monovalent salt in the secondary dialysate to obtain a second concentrated solution and a third dialysate; and introducing the second concentrated solution into an ion exchange resin system to obtain a four-stage dialysate, and extracting and crystallizing the four-stage dialysate to obtain vitamin VB 12. The method removes monovalent salt through the south-road effect of the nanofiltration membrane, reduces the load of a subsequent ion exchange resin system, obtains sodium chloride crystal salt through evaporation and crystallization after the nanofiltration membrane dialysate is concentrated, and greatly reduces the environmental protection pressure and the fresh water resource consumption.)

1. A method for separating and purifying vitamin VB12 from fermentation liquor is characterized by comprising the following steps:

s1, introducing the acidified first fermentation liquid with the pH value of 2-5 into a pretreatment system to obtain a first-stage dialysate;

s2, introducing the primary dialysate into an organic tubular membrane system to obtain a first concentrated solution and a secondary dialysate;

s3, introducing the secondary dialysate into a nanofiltration membrane system, and removing monovalent salt in the secondary dialysate to obtain a second concentrated solution and a third dialysate;

and S4, introducing the second concentrated solution into an ion exchange resin system to obtain a four-stage dialysate, and extracting and crystallizing the four-stage dialysate to obtain vitamin VB 12.

2. The method for separating and purifying vitamin VB12 from the fermentation liquid according to claim 1, wherein the first concentrated solution flows back to the pretreatment system to be mixed with the second fermentation liquid, and then the steps S2, S3 and S4 are performed in sequence.

3. The method for separating and purifying vitamin VB12 from the fermentation liquid according to claim 1, wherein the sodium chloride crystalline salt is obtained by performing an evaporation crystallization operation on the second concentrated solution.

4. The method for separating and purifying vitamin VB12 from the fermentation broth according to claim 1, wherein acetone is used for extracting the quaternary dialysate in the S4, and the extracted substances are crystallized to obtain vitamin VB 12.

5. The separation and purification method for extracting vitamin VB12 from the fermentation broth according to claim 1, wherein the functions of the pretreatment system comprise mixing, centrifugation and pressure filtration of the solution in the pretreatment system, and the equipment used by the pretreatment system comprises a disk centrifuge, a horizontal screw centrifuge, a stacked screw machine and/or a plate and frame filter press.

6. The separation and purification method for extracting vitamin VB12 from the fermentation broth according to claim 5, wherein the organic tubular membrane system comprises an organic tubular membrane, the pore diameter of the organic tubular membrane is 10 nm-100 nm, the operating pressure of the membrane system is-100-1000 k Pa, and the operating temperature is 0-40 ℃.

7. The separation and purification method for extracting vitamin VB12 from the fermentation liquor according to claim 1, wherein the nanofiltration membrane system and the organic tubular membrane system adopt an N +1 group mode, so that the cleaning mode and the working mode of each of the nanofiltration membrane system and the organic tubular membrane system are switched without affecting the treatment efficiency of each.

8. The separation and purification method for extracting vitamin VB12 from fermentation liquor according to claim 7, wherein the cleaning temperature of the organic tubular membrane system is 0-65 ℃, the working pH is 2-5, and the cleaning pH is 1-13.

9. The separation and purification method for extracting vitamin VB12 from the fermentation broth according to claim 1, wherein the nanofiltration membrane system comprises a nanofiltration membrane component, the nanofiltration membrane component can generate a Dow south effect, the nanofiltration membrane component has a negative interception effect on monovalent ions, and the interception rate on divalent ions is more than 95%.

10. A separation and purification system, comprising: the device comprises a pretreatment system, an organic tubular membrane system, a nanofiltration membrane system, an ion exchange resin system and an evaporative crystallization system;

the pretreatment system is communicated with an organic tubular membrane system, a first filtering product in the organic tubular membrane system can be led into the pretreatment system, the organic tubular membrane system is communicated with a nanofiltration membrane system, the nanofiltration membrane system is communicated with an ion exchange resin system, the nanofiltration membrane system is communicated with an evaporative crystallization system, a second filtering product in the nanofiltration membrane system is led into the evaporative crystallization system, and the evaporative crystallization system is used for evaporating and crystallizing the second filtering product to obtain crystalline salt.

Technical Field

The invention relates to the technical field of biological fermentation, in particular to a separation and purification method for extracting vitamin VB12 from fermentation liquor.

Background

VB12, namely vitamin B12 also known as cobalamin, is an important animal and human nutritional factor, is widely applied to the fields of feed, food and medicine and health, and has obvious effects in many aspects, such as promoting the development and maturation of red blood cells and preventing pernicious anemia; maintaining nervous system health; the coenzyme exists in a coenzyme form, can increase the utilization rate of folic acid and promote the metabolism of carbohydrate, fat and protein; can also promote the synthesis of protein, and has important effect on the growth and development of infants; it can also eliminate dysphoria, concentrate attention, and enhance memory and balance, and VB12 is involved in the formation of lipoprotein in nervous tissue, and is an indispensable vitamin for functional health of nervous system;

the existing VB12 production process has high energy consumption and large medicament consumption, and the generated pollutant amount is large, thus having serious influence on the environment, therefore, the invention provides a separation and purification method for extracting vitamin VB12 from fermentation liquor.

Disclosure of Invention

In order to solve the problems in the prior art, the invention provides a separation and purification method for extracting vitamin VB12 from fermentation liquor.

In order to achieve the purpose, the invention adopts the following technical scheme:

a separation and purification method for extracting vitamin VB12 from fermentation liquor comprises the following steps:

s1, introducing the acidified first fermentation liquid with the pH value of 2-5 into a pretreatment system to obtain a first-stage dialysate;

s2, introducing the primary dialysate into an organic tubular membrane system to obtain a first concentrated solution and a secondary dialysate;

s3, introducing the secondary dialysate into a nanofiltration membrane system, and removing monovalent salt in the secondary dialysate to obtain a second concentrated solution and a third dialysate;

and S4, introducing the second concentrated solution into an ion exchange resin system to obtain a four-stage dialysate, and extracting and crystallizing the four-stage dialysate to obtain vitamin VB 12.

Further, the first concentrated solution is returned to the pretreatment system to be mixed with the second fermentation solution, and then the first concentrated solution is subjected to steps S2, S3 and S4 in sequence.

Further, evaporating and crystallizing the second concentrated solution to obtain sodium chloride crystal salt.

In S4, acetone is used to extract the quaternary dialysate, and the extracted substances are crystallized to obtain vitamin VB 12.

Further, the functions of the pretreatment system comprise mixing, centrifuging and pressure filtering of the solution in the pretreatment system, and the equipment used by the pretreatment system comprises a disc centrifuge, a horizontal screw centrifuge, a screw stack machine and/or a plate and frame filter press.

Further, the organic tubular membrane system comprises an organic tubular membrane, the aperture of the organic tubular membrane is 10 nm-100 nm, the operating pressure of the membrane system is-100-1000 kPa, and the working temperature is 0-40 ℃.

Furthermore, the nanofiltration membrane system and the organic tubular membrane system adopt an N +1 group mode, so that the respective cleaning mode and working mode of the nanofiltration membrane system and the organic tubular membrane system are switched without influencing the respective treatment efficiency.

Further, the cleaning temperature of the organic tubular membrane system is 0-65 ℃, the working pH value is 2-5, and the cleaning pH value is 1-13.

Further, the nanofiltration membrane system comprises a nanofiltration membrane component, the nanofiltration membrane component can generate a south-of-the-road effect, the nanofiltration membrane component has a negative interception effect on monovalent ions, and the interception rate on divalent ions is more than 95%.

A separation and purification system comprises a pretreatment system, an organic tubular membrane system, a nanofiltration membrane system, an ion exchange resin system and an evaporative crystallization system;

the pretreatment system is communicated with an organic tubular membrane system, a first filtering product in the organic tubular membrane system can be led into the pretreatment system, the organic tubular membrane system is communicated with a nanofiltration membrane system, the nanofiltration membrane system is communicated with an ion exchange resin system, the nanofiltration membrane system is communicated with an evaporative crystallization system, a second filtering product in the nanofiltration membrane system is led into the evaporative crystallization system, and the evaporative crystallization system is used for evaporating and crystallizing the second filtering product to obtain crystalline salt.

The invention has the beneficial effects that:

1. the invention adopts a continuous system design and an N +1 group design principle, and is cleaned and reserved on line, thereby ensuring that the treatment capacity requirement can be met at any time;

2. by utilizing the cross-flow filtration principle of the organic tubular membrane, the pollution of macromolecular protein to the membrane is reduced, the high water production efficiency is ensured, the removal of the macromolecular protein can ensure the stable operation of a nanofiltration membrane system, the cleaning frequency of the nanofiltration membrane is reduced, the service time and the service life of the nanofiltration membrane are ensured, the cost of using the nanofiltration membrane system is much lower than that of a resin system, the consumption of required medicament is less than 10%, and the system occupies a small area;

3. after the organic tubular membrane is used for removing protein and the nanofiltration membrane is concentrated, the treatment amount of an ion exchange resin system is greatly reduced, the resin regeneration period is prolonged, the system occupies a small area, the medicament consumption is low, and the continuous treatment capacity is strong;

4. monovalent salt in the tertiary dialysate can obtain high-quality sodium chloride crystal salt through concentration and evaporative crystallization, has reduced salt content and the water yield of waste water, and concentrated product water and evaporation condensate water can retrieval and utilization production, and sodium chloride crystal salt can resource utilization, has alleviateed the environmental protection pressure and the water resource consumption of enterprise greatly.

Drawings

FIG. 1 is a flow chart of the separation and purification of the present invention.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments.

Referring to fig. 1, a separation and purification method for extracting vitamin VB12 from fermentation liquor comprises the following steps:

s1, acidifying the first fermentation liquid by hydrochloric acid, wherein the pH value of the acidified first fermentation liquid is 2-5, introducing the acidified first fermentation liquid into a pretreatment system, and removing large-particle suspended matters in the first fermentation liquid by utilizing pretreatment processes such as centrifugation or filter pressing to obtain a primary dialysate;

the function of the pretreatment system comprises mixing, centrifuging and filter pressing of the solution in the pretreatment system, and the equipment used by the pretreatment system comprises a disc centrifuge, a horizontal screw centrifuge, a screw stacking machine and/or a plate-and-frame filter press;

s2, introducing the primary dialysate into an organic tubular membrane system, and further removing impurities such as protein by using the organic tubular membrane system to obtain a first concentrated solution and a second dialysate;

the first concentrated solution flows back to the pretreatment system to be mixed with the second fermentation liquor, and then the first concentrated solution sequentially passes through steps S2, S3 and S4;

the organic tubular membrane system is designed for one-time continuous concentration, the first concentrated solution does not flow back to the raw water tank, is directly mixed with the second fermentation liquor and then passes through the pretreatment device, the recovery rate of the continuous concentration reaches more than 95 percent, the concentration multiple reaches more than 20 times, the one-time concentration and flow back to the pretreatment section can avoid the situation that the temperature rise of equipment operation causes the decomposition or destruction of effective substances, the design in the prior art is that the first concentrated solution flows back to the raw water tank, and the design easily causes the temperature rise to cause the destruction or decomposition of the effective substances;

the first fermentation liquid and the second fermentation liquid refer to VB12 fermentation liquid to be purified and extracted to VB12, the first fermentation liquid and the second fermentation liquid refer to the same kind of fermentation liquid, and the same kind of fermentation liquid is divided into a plurality of batches to be respectively treated;

the organic tubular membrane system comprises an organic tubular membrane, the aperture of the organic tubular membrane is 10 nm-100 nm, the operating pressure of the membrane system is-100-1000 kPa, the working temperature is 0-40 ℃, the organic tubular membrane system has a cleaning function, the cleaning temperature is 0-65 ℃, the working pH is 2-5, and the cleaning pH is 1-13;

the organic tubular membrane is a non-woven fabric supporting membrane component with an asymmetric membrane structure, and the membrane material includes but is not limited to: such as PVDF, PES, PP, PEK, PVC or PS;

by utilizing the cross-flow filtration principle of the organic tubular membrane, the pollution of macromolecular protein to the membrane is reduced, the high water production efficiency is ensured, the removal of the macromolecular protein can ensure the stable operation of a nanofiltration membrane system, the cleaning frequency of the nanofiltration membrane is reduced, the service time and the service life of the nanofiltration membrane are ensured, the cost of using the nanofiltration membrane system is much lower than that of a resin system, the consumption of required medicament is less than 10%, and the system occupies a small area;

s3, introducing the secondary dialysate into a nanofiltration membrane system, removing monovalent salt in the secondary dialysate to obtain a second concentrated solution and a third-stage dialysate, wherein the second concentrated solution contains VB12, the content of the concentrated VB12 exceeds 1mg/L, and the content of sodium chloride is less than 10000 mg/L;

the third-level dialysate contains monovalent salt with content higher than 10000 mg/L;

the nanofiltration membrane system comprises a nanofiltration membrane component, the nanofiltration membrane component can generate a south-of-the-road effect, the nanofiltration membrane component has a negative interception effect on monovalent ions, the interception rate on divalent ions is more than 95%, the nanofiltration membrane component needs to have strong acid resistance, and the nanofiltration membrane component can stably operate for a long time under the condition of a pH value of 2-3;

the equipment used by the nanofiltration membrane system comprises electrodialysis equipment and reverse osmosis membrane equipment;

monovalent salt is removed through the south-road effect of the nanofiltration membrane, and sodium chloride crystal salt can be obtained through evaporative crystallization after the third-level dialysate is concentrated;

monovalent salt in the tertiary dialysate can be concentrated and evaporated for crystallization to obtain high-quality sodium chloride crystal salt, so that the salt content and water yield of the production wastewater are reduced, the concentrated produced water and evaporated condensate water can be recycled for production, the sodium chloride crystal salt can be recycled, and the environmental protection pressure and water resource consumption of enterprises are greatly reduced;

s4, introducing the second concentrated solution into an ion exchange resin system to obtain a four-stage dialysate, extracting the four-stage dialysate, crystallizing to obtain vitamin VB12, extracting the four-stage dialysate by using acetone, and crystallizing the extracted substances to obtain vitamin VB 12;

ion exchange systems include, but are not limited to, valve matrix ion exchange resin systems, rotating disk ion exchange resin systems;

after the organic tubular membrane is used for removing protein and the nanofiltration membrane is concentrated, the treatment amount of an ion exchange resin system is greatly reduced, the resin regeneration period is prolonged, the system occupies a small area, the medicament consumption is low, and the continuous treatment capacity is strong;

performing evaporation crystallization operation on the second concentrated solution to obtain sodium chloride crystal salt, wherein equipment used for evaporation crystallization comprises but is not limited to falling film evaporation, mechanical supercharged evaporation or multi-effect evaporation, and the quality of the crystal salt meets the industrial dry salt first grade product in coal chemical industry byproduct sodium chloride T/CCT002-2019 and the refined industrial salt industrial dry salt first grade product in industrial salt sodium chloride GB/T5462-2015;

the nanofiltration membrane system and the organic tubular membrane system adopt an N +1 group mode, so that the respective cleaning mode and working mode of the nanofiltration membrane system and the organic tubular membrane system are switched without influencing the respective treatment efficiency, the online cleaning mode can realize 24h 365 day work, and the N +1 group mode design comprises that, for example, the N +1 group mode is designed into a system which originally needs 100 membranes to be formed, and the system is divided into 5 groups, then 1 group of standby systems, namely 6 groups (5+1), is added, so that the number of the membranes is 120, and 80 membranes are saved compared with a set of system (100 membranes) which is singly standby.

By adopting the design principle of N +1 group, the system can be cleaned on line and reserved, the requirement of treatment capacity can be met at any time, and the membrane system is designed in a one-step concentration manner, so that the circulating energy consumption is greatly reduced, the energy utilization rate is improved, and the product can be treated in the shortest flow;

example 1:

at a throughput of 40m³Per hour, titer 100mg/L fermentation broth for example

Acidifying raw water with hydrochloric acid, and removing large-particle suspended matters, wherein the content of monovalent salt in fermentation liquor is more than 10000mg/L, and the pH value is 2.

The flux of the organic tubular membrane is designed to be 80LMH, and the area of the organic tubular membrane required by the case is 500m²Needs 8 inches of membrane module with 6mm specification and a single membrane area of 34m^2,15 membrane modules are needed, 5 membrane modules in each group are connected in series, N +1 groups are designed, namely 4 groups of 20 membrane modules are needed, and the designed water yield is 40m³H is used as the reference value. Selecting PEK membrane material, operating pressure of 0-0.4MPa, produced water SS less than 1, SDI₁₅Less than 3, and the transmittance of 650nm is more than 90%.

The flux of the nanofiltration membrane is designed to be 15LMH, and the membrane surface is designedProduct 2667m²An 8040 membrane module is required, with a single membrane area of 27m²100 membrane modules are needed, 32 membrane modules in each group are designed, and N +1 groups are designed, namely 128 membrane modules are needed, the design is concentrated by 10 times, and the produced water is 36m³H, 4m of concentrated solution³The titer is 1000mg/L, the monovalent salt content of the nanofiltration membrane concentrated solution is less than 10000mg/L, the monovalent salt content of the secondary dialysate is more than 11000mg/L, and the operating pressure is 2-2.4 MPa.

Design ion exchange resin System throughput 4m³And h, realizing free switching between work and cleaning by utilizing a valve array type resin system, ensuring the water production efficiency to be unchanged, and realizing continuous production.

In the description of the present invention, it is to be understood that the terms "first", "second", "third", "fourth" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implying any indication of the number of technical features indicated.

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.