CN105779521A - Quick separation method for preparing L-serine crude product - Google Patents
Quick separation method for preparing L-serine crude product Download PDFInfo
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- CN105779521A CN105779521A CN201610322372.5A CN201610322372A CN105779521A CN 105779521 A CN105779521 A CN 105779521A CN 201610322372 A CN201610322372 A CN 201610322372A CN 105779521 A CN105779521 A CN 105779521A
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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
- C12P13/06—Alanine; Leucine; Isoleucine; Serine; Homoserine
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- 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
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Abstract
The invention discloses a quick separation method for preparing an L-serine crude product. The quick separation method is characterized by comprising the following steps of a, cell breaking process section; b, enzymatic synthesis process section; c, process section of separating enzymatic products from cell debris; d, preparation of the L-serine crude product. According to the quick separation method for preparing the L-serine crude product, disclosed by the invention, the preparation time of the crude product is shortened by 12 to 14 hours, and the consumption of two thirds of desorption reagent is reduced.
Description
Technical field
The present invention relates to prepare the fast separating process of Serine crude product.
Background technology
Serine is human body non essential amino acid, but owing to being in amino acid metabolism during its in vivo amino acid metabolism
Between link, participate in the synthesis of multiple important substance, therefore biological function is extremely important.At present, Serine mainly passes through silk
Propylhomoserin hydroxymethyl transferases (SHMT) Enzymatic transformation obtains.SHMT by glyA gene code, formaldehyde, pyridoxal 5-phosphate and
Tetrahydrofolic acid can be catalyzed glycine under conditions of existing and be converted into Serine.Utilize Enzymatic transformation to produce Serine and have former
Material price is low, process route is short, reaction time is fast, product assay high.But prepared serine afterproduct by enzyme process
The isolation and purification always people problem that compares concern.Wei Pinghe et al. is " separation of Production by Enzymes Serine is pure
Change " one the article pointed out enzyme process to prepare in the separation purifying technique of serine to separate cell debris and albumen uses perlite drainage, nothing
The methods such as machine flocculant precipitating proteins and activated carbon removing pigment, the stillness of night after separation is again with ultrafiltration-nanofiltration, reverse osmosis
Through resins exchange separation product after process.Before this, also there is document report as " in SHM T enzymatic reaction solution, Serine divides
From with relevant component analysis ", " amino acid whose separation in enzymatic clarification Serine and reactant liquor " propose with 717 the moon from
The method of subtree fat separation Serine.In other amino acid whose separation processes, utilize ion exchange resin amino acid separation
Also it is a kind of method of conventional separation for amino acids.But it is either amino acid whose in the separation of Enzymatic transformation product or tunning
Separate, utilize resins exchange to obtain aminoacid and all there is the spies such as separation cycle length, strippant consumption is big, environmental pollution is serious
Point.Therefore, set up a kind of simple product separation method and there is extraordinary application prospect.
Summary of the invention
The problem to be solved in the present invention is to provide a kind of fast separating process preparing Serine crude product.
A kind of fast separating process preparing Serine crude product of the present invention, it is characterised in that include following steps: a. cell breakage
Process section: being resuspended in deionized water by fermentation gained bacterium 28-35kg, cumulative volume is 400L, is added thereto to CTAB broken thin
Born of the same parents, CTAB addition is total volume concentration 0.05-0.2%；B. Enzymatic transformation process section: above-mentioned broken cell mixture is pumped into
In conversion tank, adding deionized water is 600L to cumulative volume, glycine 2-2.8mol/L, PLP 0.2-0.4 in conversion tank
Mmol/L, THFA3-5mmol/L, formaldehyde 8-13mmol/L, pH value is 6.0-8.0, stirs 100-150rpm, temperature 35-
40 DEG C, the time is 35-50h, reaches enzyme equilibrium；C. separation enzymatic preparation and cell debris process section: Enzymatic transformation
Mixed liquor after balance adds water to 1000-1500L membrane aperture 0.2～80 μm chimney filter ceramic membrane filter and removes thalline；d.
Prepared by Serine crude product: the reactant liquor of enzymatic reaction removes thalline through ceramic membrane filter, collects reactant liquor and is proceeded to subtracting
Pressure distillation column in concentrating under reduced pressure, vacuum is 0.07-0.09Mpa, and temperature is 30 DEG C to 50 DEG C, be concentrated into volume for concentrate before
The 1/4-1/6 of volume, precipitated in a large number, be Serine crude product, time-consuming 4-8 hour；Remaining 1/4-1/6 body
Long-pending concentrated solution prepared Serine crude product according to former technique crystallization after repeatedly collecting.
The former method preparing Serine crude product includes following steps: a. cell breakage process section: will fermentation gained bacterium 28-
35kg is resuspended in deionized water, and cumulative volume is 400L, is added thereto to CTAB broken cell, and CTAB addition is cumulative volume
Concentration 0.05-0.2%；B. cell debris is separated and to separating liquid concentration technique section: after broken cell, mixeding liquid volume adds
1000-1500L deionized water, and separate after being added thereto to seprating assistant thing, vacuum filtration, obtain separating liquid 1200-
1700L；At 35-40 DEG C, separation liquid is concentrated in vacuo to volume is 400L；C. Enzymatic transformation process section: above-mentioned broken cell is mixed
Compound pumps in conversion tank, and adding deionized water is 600L to cumulative volume, glycine 2-2.8mol/L, PLP in conversion tank
0.2-0.4mmol/L, THFA3-5mmol/L, formaldehyde 8-13mmol/L, pH value is 6.0-8.0, stirs 100-150rpm,
Temperature 35-40 DEG C, the time is 35-50h, reaches enzyme equilibrium；D. post separating technology section: to enzyme equilibrium liquid
Middle addition deionized water 1000-1500L upper 717 anion columns after adjusting pH to 7.0-7.5, use after loading
The hydrochloric acid eluting of 0.15mol/L, obtains containing Serine eluent 1000-1500L；E. crystallization processes section: by Serine
Eluent be evaporated to 300L at 50-60 DEG C, and be added thereto to 600-900L dehydrated alcohol 5-10 DEG C of crystallization,
To Serine.
The present invention is directed to Serine Enzymatic transformation efficiency and the high feature of production concentration, devise and a kind of separate Serine
The fast method of crude product, shortened the time preparing crude product 12-14 hour, decreased the use of the strippant of 2/3.
Detailed description of the invention
Embodiment 1: a kind of fast separating process preparing Serine crude product, includes following steps: a. cell breakage technique
Section: being resuspended in deionized water by fermentation gained bacterium 28-35kg, cumulative volume is 400L, is added thereto to CTAB broken cell,
CTAB addition is total volume concentration 0.05-0.2%；B. Enzymatic transformation process section: above-mentioned broken cell mixture is pumped into conversion tank
In, adding deionized water is 600L to cumulative volume, glycine 2-2.8mol/L, PLP 0.2-0.4mmol/L in conversion tank,
THFA3-5mmol/L, formaldehyde 8-13mmol/L, pH value is 6.0-8.0, stirs 100-150rpm, temperature 35-40 DEG C, time
For 35-50h, reach enzyme equilibrium；C. separation enzymatic preparation and cell debris process section: mixing after Enzymatic transformation balance
Close liquid and add water to 1000-1500L membrane aperture 0.2～80 μm chimney filter ceramic membrane filter removal thalline；D.L-serine is thick
Prepared by product: the reactant liquor of enzymatic reaction removes thalline through ceramic membrane filter, collects reactant liquor and is proceeded to vacuum distillation tower
Concentrating under reduced pressure, vacuum is 0.07-0.09Mpa, and temperature is 30 DEG C to 50 DEG C, and being concentrated into volume is the volume before concentrating
1/4-1/6, is precipitated in a large number, is Serine crude product, time-consuming 4-8 hour；The concentration of remaining 1/4-1/6 volume
Liquid prepared Serine crude product according to former technique crystallization after repeatedly collecting.
Embodiment 2: fermentation gained bacterium 28-35kg is resuspended in deionized water (cumulative volume is 400L), is added thereto to
CTAB is to final concentration of 0.05-0.2% broken cell.After cell breakage after broken cell in mixed liquor (volume is about 400L)
Adding deionized water to cumulative volume is 600 liters, the wherein 2-2.8mol/L Han glycine, PLP 0.2-0.4mmol/L,
THFA3-5mmol/L, formaldehyde 8-13mmol/L, pH:6.0-8.0, stir 100-150rpm, temperature 35-40 DEG C, reach
Enzyme equilibrium.Balancing response liquid filters through ceramic membrane (membrane aperture 0.2～80 μm chimney filter) and removes thalline, collects
Reactant liquor is also proceeded to vacuum distillation tower concentrating under reduced pressure, and vacuum is 0.07Mpa, and temperature is 40 DEG C, is concentrated into volume
For 1/4 of the volume before concentrating, obtain 45kg precipitation, precipitate is carried out detection and proves Serine, be L-silk ammonia
Acid crude, time-consuming 5-5.5 hour.
Embodiment 3: fermentation gained bacterium 28-35kg is resuspended in deionized water (cumulative volume is 400L), is added thereto to
CTAB is to final concentration of 0.05-0.2% broken cell.After cell breakage after broken cell in mixed liquor (volume is about 400L)
Adding deionized water to cumulative volume is 600 liters, the wherein 2-2.8mol/L Han glycine, PLP 0.2-0.4mmol/L,
THFA3-5mmol/L, formaldehyde 8-13mmol/L, pH:6.0-8.0, stir 100-150rpm, temperature 35-40 DEG C, reach
Enzyme equilibrium.Balancing response liquid filters through ceramic membrane (membrane aperture 0.2～80 μm chimney filter) and removes thalline, collects
Reactant liquor is also proceeded to vacuum distillation tower concentrating under reduced pressure, and vacuum is 0.07Mpa, and temperature is 40 DEG C, is concentrated into volume
For 1/6 of the volume before concentrating, obtain 59.5kg precipitation, precipitate is carried out detection and proves Serine, without sweet ammonia
Acid, is Serine crude product, time-consuming 5.5-6 hour.
Embodiment 4: fermentation gained bacterium 28-35kg is resuspended in deionized water (cumulative volume is 400L), is added thereto to
CTAB is to final concentration of 0.05-0.2% broken cell.After cell breakage after broken cell in mixed liquor (volume is about 400L)
Adding deionized water to cumulative volume is 600 liters, the wherein 2-2.8mol/L Han glycine, PLP 0.2-0.4mmol/L,
THFA3-5mmol/L, formaldehyde 8-13mmol/L, pH:6.0-8.0, stir 100-150rpm, temperature 35-40 DEG C, reach
Enzyme equilibrium.Balancing response liquid filters through ceramic membrane (membrane aperture 0.2～80 μm chimney filter) and removes thalline, collects
Reactant liquor is also proceeded to vacuum distillation tower concentrating under reduced pressure, and vacuum is 0.07Mpa, and temperature is 50 DEG C, is concentrated into volume
For 1/6 of the volume before concentrating, obtain 60kg precipitation, precipitate is carried out detection and proves Serine, without sweet ammonia
Acid, is Serine crude product, time-consuming 5.5-6 hour.
Embodiment 5: fermentation gained bacterium 28-35kg is resuspended in deionized water (cumulative volume is 400L), is added thereto to
CTAB is to final concentration of 0.05-0.2% broken cell.After cell breakage after broken cell in mixed liquor (volume is about 400L)
Adding deionized water to cumulative volume is 600 liters, the wherein 2-2.8mol/L Han glycine, PLP 0.2-0.4mmol/L,
THFA3-5mmol/L, formaldehyde 8-13mmol/L, pH:6.0-8.0, stir 100-150rpm, temperature 35-40 DEG C, reach
Enzyme equilibrium.Balancing response liquid filters through ceramic membrane (membrane aperture 0.2～80 μm chimney filter) and removes thalline, collects
Reactant liquor is also proceeded to vacuum distillation tower concentrating under reduced pressure, and vacuum is 0.09Mpa, and temperature is 50 DEG C, is concentrated into volume
For 1/6 of the volume before concentrating, obtain 60kg precipitation, precipitate is carried out detection and proves Serine, without sweet ammonia
Acid, is Serine crude product, time-consuming 4-5 hour.
Claims (1)
1. the fast separating process preparing Serine crude product, it is characterised in that include following steps: a. cell breakage
Process section: being resuspended in deionized water by fermentation gained bacterium 28-35kg, cumulative volume is 400L, is added thereto to CTAB
Broken cell, CTAB addition is total volume concentration 0.05-0.2%；B. Enzymatic transformation process section: above-mentioned broken cell is mixed
Thing pumps in conversion tank, and adding deionized water is 600L to cumulative volume, glycine 2-2.8mol/L, PLP in conversion tank
0.2-0.4mmol/L, THFA3-5mmol/L, formaldehyde 8-13mmol/L, pH value is 6.0-8.0, stirs 100-
150rpm, temperature 35-40 DEG C, the time is 35-50h, reaches enzyme equilibrium；C. enzymatic preparation is separated broken with cell
Blade technolgy section: the mixed liquor after Enzymatic transformation balance adds water to 1000-1500L membrane aperture 0.2～80 μm chimney filter pottery
Porcelain membrane filtration removes thalline；Prepared by d.L-serine crude product: the reactant liquor of enzymatic reaction removes thalline through ceramic membrane filter,
Collecting reactant liquor and proceeded to vacuum distillation tower concentrating under reduced pressure, vacuum is 0.07-0.09Mpa, and temperature is 30 DEG C
To 50 DEG C, it is concentrated into the 1/4-1/6 that volume is the volume before concentrating, is precipitated in a large number, be Serine crude product,
Time-consuming 4-8 hour；The concentrated solution of remaining 1/4-1/6 volume prepares L-silk ammonia according to the crystallization of former technique after repeatedly collecting
Acid crude.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610322372.5A CN105779521A (en) | 2016-05-12 | 2016-05-12 | Quick separation method for preparing L-serine crude product |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610322372.5A CN105779521A (en) | 2016-05-12 | 2016-05-12 | Quick separation method for preparing L-serine crude product |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110205346A (en) * | 2019-07-02 | 2019-09-06 | 武汉轻工大学 | A kind of preparation method of Serine |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101787356A (en) * | 2010-02-03 | 2010-07-28 | 湖北省八峰药化股份有限公司 | Construction and immobilized use method of glyA genetically engineered microorganism |
| CN102220389A (en) * | 2011-04-20 | 2011-10-19 | 横店集团家园化工有限公司 | Synthetic method of L-serine |
| CN104946695A (en) * | 2015-06-03 | 2015-09-30 | 武汉轻工大学 | Preparation method of L-serine |
-
2016
- 2016-05-12 CN CN201610322372.5A patent/CN105779521A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101787356A (en) * | 2010-02-03 | 2010-07-28 | 湖北省八峰药化股份有限公司 | Construction and immobilized use method of glyA genetically engineered microorganism |
| CN102220389A (en) * | 2011-04-20 | 2011-10-19 | 横店集团家园化工有限公司 | Synthetic method of L-serine |
| CN104946695A (en) * | 2015-06-03 | 2015-09-30 | 武汉轻工大学 | Preparation method of L-serine |
Non-Patent Citations (2)
| Title |
|---|
| HUMG-YU HSIAO: "Enzymatic Production of L-Serine with a Feedback Control System for Formaldehyde Addition", 《BIOTECHNOLOGY AND BIOENGINEERING》 * |
| 赵黎明主编: "《中国纺织出版社》", 31 July 2011 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110205346A (en) * | 2019-07-02 | 2019-09-06 | 武汉轻工大学 | A kind of preparation method of Serine |
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Application publication date: 20160720 |
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