CN110872606A - Preparation method of L-lysine sulfate and by-product thereof - Google Patents

Preparation method of L-lysine sulfate and by-product thereof Download PDF

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CN110872606A
CN110872606A CN201910824819.2A CN201910824819A CN110872606A CN 110872606 A CN110872606 A CN 110872606A CN 201910824819 A CN201910824819 A CN 201910824819A CN 110872606 A CN110872606 A CN 110872606A
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lysine
fermentation
sulfate
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producing
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王成福
唐瑞
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Heilongjiang Fortune Food Group Co Ltd
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P13/00Preparation of nitrogen-containing organic compounds
    • C12P13/04Alpha- or beta- amino acids
    • C12P13/08Lysine; Diaminopimelic acid; Threonine; Valine
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    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C227/00Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C227/38Separation; Purification; Stabilisation; Use of additives
    • C07C227/40Separation; Purification
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07BGENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
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    • C07B2200/07Optical isomers

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Abstract

The invention discloses a preparation method of L-lysine sulfate and a byproduct thereof, belonging to the technical field of lysine production process and comprising the following steps: adjusting the pH of a fermentation culture medium to 7.0-7.1 by using 20% NaOH, and then continuously heating the culture medium by using steam to cause the death of microorganisms; inoculating lysine producing bacteria under sterile condition, controlling temperature at 35-40 deg.C, pH at 6.8-7.0 and DO at 20-30% during fermentation, and fermenting for 48-60 hr; after the fermentation is finished, putting the mixture of lysine and the substrate in an isoelectric extraction workshop, adjusting the pH value of the fermentation solution to 5-6 by using sulfuric acid, filtering the treated fermentation solution by ceramic membrane equipment, performing triple-effect concentration on the filtered clear solution, and performing granulation drying by using a spray granulation bed to obtain a finished product. The invention overcomes the difficult problem of environmental protection, reduces the production links, has short process route, low consumption of raw and auxiliary materials and low cost, and has high content of produced lysine.

Description

Preparation method of L-lysine sulfate and by-product thereof
Technical Field
The invention relates to the technical field of lysine production processes, and particularly relates to a preparation method of L-lysine sulfate and a by-product thereof.
Background
Lysine (Lysine) has the chemical name 2, 6-diaminohexanoic acid. Lysine is a basic essential amino acid. The cereal is called the first limiting amino acid because of its very low lysine content and its easy destruction and lack during processing.
Lysine is one of the essential amino acids in humans and mammals, and is not synthesized by the body itself and must be supplemented from food. Lysine is mainly present in animal foods and legumes, and lysine content in cereals is very low. Lysine has positive nutritional significance in promoting growth and development of human body, enhancing immunity of organism, resisting virus, promoting fat oxidation, relieving anxiety, promoting absorption of certain nutrients, and can act synergistically with certain nutrients to better exert physiological functions of various nutrients.
The direct fermentation method is a widely used lysine production method. The commonly used raw materials are cheap sugar raw materials such as waste molasses, starch hydrolysate and the like after sugar production of sugarcane or beet. In addition, acetic acid, ethanol, etc. are also optional raw materials. The main microorganisms for producing lysine by direct fermentation method include mutant strains of Corynebacterium glutamicum, Brevibacterium flavum and Brevibacterium lactofermentum. The method is developed in the later 50 th of the 20 th century, and since 70 s, some mutant strains with multiple genetic markers are bred due to the development of breeding technology, so that the process is mature day by day, and the yield of lysine is increased manyfold. The maximum yield of acid in industrial production is improved to 120g per liter of fermentation, and the extraction rate reaches about 80-90%.
Lysine, one of the feed additive materials, plays an extremely important role in feed formulation and nutrient supply. At present, 98 lysine hydrochloride and 70 lysine sulfate are taken as main components in lysine products in markets at home and abroad. 98 lysine hydrochloride, which is produced by ion exchange, has great environmental pollution and high production cost.
Disclosure of Invention
The invention provides a preparation method of L-lysine sulfate and a byproduct thereof, which solves the problems of great environmental pollution and high production cost of the existing direct fermentation production process.
In order to solve the technical problems, the invention is realized by the following technical scheme:
the invention provides a preparation method of L-lysine sulfate and a byproduct thereof, which comprises the following steps:
(1) and (3) sterilizing a culture medium: taking a fermentation medium, wherein the fermentation medium comprises the following components: 150g/L glucose, 100g/L molasses, 15g/L corn steep liquor, 50g/L ammonium sulfate, 1g/L phosphoric acid, 1.5g/L potassium chloride, 0.5g/L magnesium sulfate, 0.01g/L ferrous sulfate, 0.1g/L, VB1200 mu g/L manganese sulfate and 40g/L calcium carbonate; adjusting the pH of a fermentation medium to 7.0-7.1 by using 20% NaOH, and then continuously heating the fermentation medium by using steam to cause the death of microorganisms; by adopting the fermentation medium, the acid production rate of lysine is further improved;
(2) fermentation: inoculating lysine producing bacteria into a fermentation culture medium under an aseptic condition, controlling the fermentation process at 35-40 ℃, pH value of 6.8-7.0 and DO value of 20-30%, and allowing the lysine producing bacteria to maximally accumulate lysine through a metabolic process when the fermentation time is 48-60 hours;
(3) extraction and purification: after fermentation, putting the mixture of lysine and substrate in an isoelectric extraction workshop, adjusting the pH value of fermentation liquid to 5-6 by using sulfuric acid, filtering the treated fermentation liquid by ceramic membrane equipment to remove bacteria, viscous substances and residues, performing triple-effect concentration on the filtered clear liquid to obtain concentrated liquid with the concentration of 20 Baume, and performing granulation drying by using a spray granulation bed to obtain a finished product.
Wherein, the temperature of the steam in the step (1) is preferably 120-130 ℃.
Among them, preferably, the lysine-producing bacterium is Corynebacterium glutamicum.
Wherein, the inoculation amount of the lysine-producing bacteria is preferably 15-18% of the weight of the culture medium.
Wherein, preferably: the aperture of the ceramic membrane is 50 nm.
Compared with the prior art, the technical scheme of the invention has the following obvious effects:
in the extraction and purification process, after the pH value is adjusted by sulfuric acid, a ceramic membrane device is adopted for filtering to remove thalli, sticky substances and residues, and the filtered clear solution is subjected to triple-effect concentration.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.
Example 1
This example provides a process for the preparation of L-lysine sulfate and its by-products comprising the steps of:
(1) and (3) sterilizing a culture medium: taking a fermentation medium, wherein the fermentation medium comprises the following components: 150g/L glucose, 100g/L molasses, 15g/L corn steep liquor, 50g/L ammonium sulfate, 1g/L phosphoric acid, 1.5g/L potassium chloride, 0.5g/L magnesium sulfate, 0.01g/L ferrous sulfate, 0.1g/L, VB1200 mu g/L manganese sulfate and 40g/L calcium carbonate; before use, the pH of a fermentation medium is adjusted to 7.0 by using 20% NaOH, and then the fermentation medium is continuously heated by using steam at 125 ℃ so as to cause the death of microorganisms;
(2) fermentation: inoculating corynebacterium glutamicum into a fermentation medium under an aseptic condition, wherein the inoculation amount is 17% of the weight of the medium, the fermentation process is controlled at 35-40 ℃, the pH value is 6.9, the DO value is 25%, and the fermentation time is 54 hours, so that the corynebacterium glutamicum can maximally accumulate lysine through a metabolic process;
(3) extraction and purification: after fermentation, putting the mixture of lysine and substrate in an isoelectric extraction workshop, adjusting the pH value of the fermentation solution to 5.5 by using sulfuric acid, filtering the treated fermentation solution by using a ceramic membrane device with the pore diameter of 50nm to remove thalli, sticky substances and residues, performing triple-effect concentration on the filtered clear solution to obtain a concentrated solution with the concentration of 20 Baume, and performing granulation drying by using a spray granulation bed to obtain a finished product.
Example 2
This example provides a process for the preparation of L-lysine sulfate and its by-products comprising the steps of:
(1) and (3) sterilizing a culture medium: taking a fermentation medium, wherein the fermentation medium comprises the following components: 150g/L glucose, 100g/L molasses, 15g/L corn steep liquor, 50g/L ammonium sulfate, 1g/L phosphoric acid, 1.5g/L potassium chloride, 0.5g/L magnesium sulfate, 0.01g/L ferrous sulfate, 0.1g/L, VB1200 mu g/L manganese sulfate and 40g/L calcium carbonate; before use, the pH of a fermentation medium is adjusted to 7.0 by using 20% NaOH, and then the fermentation medium is continuously heated by using steam at 130 ℃ so as to cause the death of microorganisms;
(2) fermentation: inoculating corynebacterium glutamicum into a fermentation medium under an aseptic condition, wherein the inoculation amount is 15% of the weight of the medium, the temperature is controlled to be 35-40 ℃, the pH value is 7.0, the DO value is 20% in the fermentation process, and the fermentation time is 60 hours, so that the corynebacterium glutamicum can maximally accumulate lysine in the metabolic process;
(3) extraction and purification: after the fermentation is finished, putting the mixture of lysine and a substrate in an isoelectric extraction workshop, adjusting the pH value of a fermentation solution to 5 by using sulfuric acid, filtering the treated fermentation solution by using ceramic membrane equipment with the pore diameter of 50nm to remove thalli, sticky substances and residues, performing triple-effect concentration on the filtered clear solution to obtain a concentrated solution with the concentration of 20 Baume, and performing granulation drying by using a spray granulation bed to obtain a finished product.
Example 3
This example provides a process for the preparation of L-lysine sulfate and its by-products comprising the steps of:
(1) and (3) sterilizing a culture medium: taking a fermentation medium, wherein the fermentation medium comprises the following components: 150g/L glucose, 100g/L molasses, 15g/L corn steep liquor, 50g/L ammonium sulfate, 1g/L phosphoric acid, 1.5g/L potassium chloride, 0.5g/L magnesium sulfate, 0.01g/L ferrous sulfate, 0.1g/L, VB1200 mu g/L manganese sulfate and 40g/L calcium carbonate; before use, the pH of a fermentation medium is adjusted to 7.1 by using 20% NaOH, and then the fermentation medium is continuously heated by using steam at 120 ℃ so as to cause the death of microorganisms;
(2) fermentation: inoculating corynebacterium glutamicum into a fermentation medium under an aseptic condition, wherein the inoculation amount is 18% of the weight of the medium, the fermentation process is controlled at 35-40 ℃, the pH value is 6.8, the DO value is 30%, and the fermentation time is 48 hours, so that the corynebacterium glutamicum can maximally accumulate lysine through a metabolic process;
(3) extraction and purification: after the fermentation is finished, putting the mixture of lysine and a substrate in an isoelectric extraction workshop, adjusting the pH value of a fermentation solution to 6 by using sulfuric acid, filtering the treated fermentation solution by using ceramic membrane equipment with the pore diameter of 50nm to remove thalli, sticky substances and residues, performing triple-effect concentration on the filtered clear solution to obtain a concentrated solution with the concentration of 20 Baume, and performing granulation drying by using a spray granulation bed to obtain a finished product.
Example 4
This example provides a process for the preparation of L-lysine sulfate and its by-products comprising the steps of:
(1) and (3) sterilizing a culture medium: taking a fermentation medium, wherein the fermentation medium comprises the following components: 150g/L glucose, 100g/L molasses, 15g/L corn steep liquor, 50g/L ammonium sulfate, 1g/L phosphoric acid, 1.5g/L potassium chloride, 0.5g/L magnesium sulfate, 0.01g/L ferrous sulfate, 0.1g/L, VB1200 mu g/L manganese sulfate and 40g/L calcium carbonate; before use, the pH of a fermentation medium is adjusted to 7.1 by using 20% NaOH, and then the fermentation medium is continuously heated by using steam at 130 ℃ so as to cause the death of microorganisms;
(2) fermentation: inoculating corynebacterium glutamicum into a fermentation medium under an aseptic condition, wherein the inoculation amount is 17% of the weight of the medium, the fermentation process is controlled at 35-40 ℃, the pH value is 6.9, the DO value is 28%, and the fermentation time is 55 hours, so that the corynebacterium glutamicum can maximally accumulate lysine through a metabolic process;
(3) extraction and purification: after fermentation, putting the mixture of lysine and substrate in an isoelectric extraction workshop, adjusting the pH value of the fermentation solution to 5.4 by using sulfuric acid, filtering the treated fermentation solution by using a ceramic membrane device with the pore diameter of 50nm to remove thalli, sticky substances and residues, performing triple-effect concentration on the filtered clear solution to obtain a concentrated solution with the concentration of 20 Baume, and performing granulation drying by using a spray granulation bed to obtain a finished product.
Example 5
This example provides a process for the preparation of L-lysine sulfate and its by-products comprising the steps of:
(1) and (3) sterilizing a culture medium: taking a fermentation medium, wherein the fermentation medium comprises the following components: 150g/L glucose, 100g/L molasses, 15g/L corn steep liquor, 50g/L ammonium sulfate, 1g/L phosphoric acid, 1.5g/L potassium chloride, 0.5g/L magnesium sulfate, 0.01g/L ferrous sulfate, 0.1g/L, VB1200 mu g/L manganese sulfate and 40g/L calcium carbonate; before use, the pH of a fermentation medium is adjusted to 7.1 by using 20% NaOH, and then the fermentation medium is continuously heated by using steam at 125 ℃ so as to cause the death of microorganisms;
(2) fermentation: inoculating corynebacterium glutamicum into a fermentation medium under an aseptic condition, wherein the inoculation amount is 16% of the weight of the medium, the fermentation process is controlled at 35-40 ℃, the pH value is 6.9, the DO value is 24%, and the fermentation time is 52 hours, so that the corynebacterium glutamicum can maximally accumulate lysine through a metabolic process;
(3) extraction and purification: after fermentation, putting the mixture of lysine and substrate in an isoelectric extraction workshop, adjusting the pH value of the fermentation solution to 5.8 by using sulfuric acid, filtering the treated fermentation solution by using a ceramic membrane device with the pore diameter of 50nm to remove thalli, sticky substances and residues, performing triple-effect concentration on the filtered clear solution to obtain a concentrated solution with the concentration of 20 Baume, and performing granulation drying by using a spray granulation bed to obtain a finished product.
The performance indicators obtained in examples 1-5 were examined and the results were as follows:
Figure BDA0002188743320000051
although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.

Claims (5)

  1. A process for the preparation of L-lysine sulphate and its by-products, characterised in that it comprises the steps of:
    (1) and (3) sterilizing a culture medium: taking a fermentation medium, wherein the fermentation medium comprises the following components: 150g/L glucose, 100g/L molasses, 15g/L corn steep liquor, 50g/L ammonium sulfate, 1g/L phosphoric acid, 1.5g/L potassium chloride, 0.5g/L magnesium sulfate, 0.01g/L ferrous sulfate, 0.1g/L, VB1200 mu g/L manganese sulfate and 40g/L calcium carbonate; adjusting the pH of a fermentation culture medium to 7.0-7.1 by using 20% NaOH, and then continuously heating the culture medium by using steam to cause the death of microorganisms;
    (2) fermentation: inoculating lysine producing bacteria into a fermentation culture medium under an aseptic condition, controlling the temperature to be 35-40 ℃, the pH value to be 6.8-7.0 and the DO value to be 20-30% in the fermentation process, and fermenting for 48-60 hours to ensure that the lysine producing bacteria maximally accumulate lysine through the metabolic process;
    (3) extraction and purification: after fermentation, putting the mixture of lysine and substrate in an isoelectric extraction workshop, adjusting the pH value of fermentation liquid to 5-6 by using sulfuric acid, filtering the treated fermentation liquid by ceramic membrane equipment to remove thalli, sticky substances and residues, performing triple-effect concentration on the filtered clear liquid to obtain a concentrated solution with the concentration of 20 Baume, and performing granulation drying by using a spray granulation bed to obtain a finished product.
  2. 2. The method of producing L-lysine sulfate and by-products thereof according to claim 1, characterized in that: the temperature of the steam in the step (1) is 120-130 ℃.
  3. 3. The method of producing L-lysine sulfate and by-products thereof according to claim 1, characterized in that: the lysine producing strain is Corynebacterium glutamicum.
  4. 4. A method of producing L-lysine sulfate and by-products thereof according to claim 3, characterized in that: the inoculation amount of the lysine producing bacteria is 15-18% of the weight of the culture medium.
  5. 5. The method of producing L-lysine sulfate and by-products thereof according to claim 1, characterized in that: the aperture of the ceramic membrane is 50 nm.
CN201910824819.2A 2019-09-02 2019-09-02 Preparation method of L-lysine sulfate and by-product thereof Pending CN110872606A (en)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102442919A (en) * 2010-10-13 2012-05-09 中粮生物化学(安徽)股份有限公司 Method for producing lysine product
CN102533891A (en) * 2012-01-13 2012-07-04 中粮生物化学(安徽)股份有限公司 Production method of lysine
CN103553951A (en) * 2013-10-31 2014-02-05 安徽丰原发酵技术工程研究有限公司 Method of extracting and preparing lysine sulphate from fermenting liquid containing lysin
CN104726510A (en) * 2013-12-24 2015-06-24 中粮生物化学(安徽)股份有限公司 Method for preparing lysine through fermenting
CN106755156A (en) * 2016-12-28 2017-05-31 安徽丰原发酵技术工程研究有限公司 A kind of fermentation process of L lysines

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102442919A (en) * 2010-10-13 2012-05-09 中粮生物化学(安徽)股份有限公司 Method for producing lysine product
CN102533891A (en) * 2012-01-13 2012-07-04 中粮生物化学(安徽)股份有限公司 Production method of lysine
CN103553951A (en) * 2013-10-31 2014-02-05 安徽丰原发酵技术工程研究有限公司 Method of extracting and preparing lysine sulphate from fermenting liquid containing lysin
CN104726510A (en) * 2013-12-24 2015-06-24 中粮生物化学(安徽)股份有限公司 Method for preparing lysine through fermenting
CN106755156A (en) * 2016-12-28 2017-05-31 安徽丰原发酵技术工程研究有限公司 A kind of fermentation process of L lysines

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