CN106086101A - A kind of fermentation technology of L tryptophan - Google Patents
A kind of fermentation technology of L tryptophan Download PDFInfo
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- CN106086101A CN106086101A CN201610712487.5A CN201610712487A CN106086101A CN 106086101 A CN106086101 A CN 106086101A CN 201610712487 A CN201610712487 A CN 201610712487A CN 106086101 A CN106086101 A CN 106086101A
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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
- C12P13/22—Tryptophan; Tyrosine; Phenylalanine; 3,4-Dihydroxyphenylalanine
- C12P13/227—Tryptophan
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Abstract
The invention provides the fermentation technology of a kind of L tryptophan, the cottonseed meal adding 2g/L in L tryptophan fermentation medium ferments；Sweat is circulated filtration when yield of acetic acid is more than 5g/L, and the fermentation clear liquid after separation enters abstraction process；Concentration bacterium solution after extraction continues ferment and get final product after adding Dialysis culture base after entering fermentation tank；Described fermentation technology fermentation L tryptophan produces acid cycle stretch-out 20h, and acetic acid concentration maintains below 5g/L, single tank output increased more than 89%, and conversion ratio is improved to more than 18.5% by 17.0%；Owing to adding cottonseed meal in fermentation medium, the cotton seed hulls powder not utilized by thalline plays filtrating aid function in filtration, and filtering fermentation liquor speed is promoted to 183mL/L by 85mL/min.
Description
Technical field
The present invention relates to tryptophan production field, the fermentation technology of a kind of L-Trp.
Background technology
L-Trp has another name called L-pantonine-indolylalanine, chemical name L-2-amino-3-indolylalanine, is
Containing the neutral aromatic amino acid of indyl, play an important role in the metabolism of humans and animals body, be humans and animals
One of 8 kinds of internal essential amino acids.The physical property of L-Trp is white or the leaflet crystal of yellowish or powder
End, odorless or micro-smelly, pleasantly sweet, it is dissolved in hot pyridine, is slightly soluble in ethanol insoluble in chloroform and ether.L-Trp heats meeting with water
Having a small amount of indole to produce, under conditions of with the presence of NaOH, CuSO4, heating has the generation of substantial amounts of indole.At present, L-Trp
It is widely used in the aspects such as medicine, food and feedstuff.
In terms of medicine, before the L-Trp synthesis hormone such as serotonine, heteroauxing and biological active substances
Body thing.Tryptophan also maintenance to brain function activity plays an important role, and when human body lacks tryptophan, can produce neural wrong
Random hallucination, stress reduces, and attention is the most corresponding to memory to go down.L-Trp is treated as medicine at present
Pellagra, schizophrenia.In addition, L-Trp also has promotion sleep quality, and resisting hypertension and analgesia etc. act on.
In terms of food, due to L-Trp can and metal ion-chelant, therefore it can prevent albumen as preservative
Matter is gone mouldy reaction.L-Trp and Ala-Trp dipeptides etc. have the aminoacid use in conjunction tool of anti-oxidation function
There is higher antisepsis.6-chlorine tryptophan is a derivant of D-trp, and its sugariness is 1300 times of sweetness of cane sugar, therefore
Tryptophan can also be taken by obesity, hypertension and diabetics as the sweeting agent of non-nutritive.Further, since plant
Thing albumen compares shortage tryptophan, can significantly improve the utilization rate of vegetable protein with tryptophan condensed food and feedstuff.
In terms of feedstuff, so far, L-Trp is confirmed to be six kinds of feedstuffs and adds one of aminoacid, becomes the bad ammonia that continues
The 3rd class feedstuff after acid and methionine adds aminoacid.Tryptophan can improve the synthetic ratio of animal body protein, and
Niacin amide can be converted into and promote the growth of animal body.And L-Trp can improve the conjunction of gamma globulin in animal body
Cheng Liang, improves the resistance against diseases of animal body.L-Trp also can produce certain impact to the behavior of animal, adds in feedstuff
A certain amount of tryptophan can significantly decrease the aggressive behavior of animal, reduces the generation of stress.
So far, the production method of L-Trp has enzyme process, microbe transformation method and microbe fermentation method.But due to
During enzymatic clarification tryptophan, the requirement of enzyme is big and easy in inactivation, and enzyme is expensive, exists in commercial production
The biggest drawback.Precursor substance required in microbe transformation method inherently growth to microorganism serves inhibitory action,
Commercial production also receives and greatly limits.And microbe fermentation method has the excellent of the uniquenesses such as product acid height, low cost, quality are good
Gesture, becomes the one preferred technique of large-scale production tryptophan.
During Production by Microorganism Fermentation tryptophan, in order to improve the yield of L-Trp, main employing optimizes training
Support base and add the mode of culture.Huang Jianmin et al. improves yield (patent No. ZL of tryptophan by adding Sodium Pyruvate
201310692893.6).Li Rongjie et al. improves tryptophan yield (specially by adding NPE-10 (OP-10)
Profit ZL 201110034187.3).But, use the mode adding culture and Optimal Medium to improve the product of tryptophan
Amount, it is impossible to enough solve fermentation middle and late stage Acetic Acid Accumulation suppression and produce the phenomenon of acid, and add nutrient and make the composition of fermentation liquid become
Obtain increasingly complex, cause certain difficulty to the separation and Extraction process in later stage.
Summary of the invention
The technical problem to be solved is to provide the fermentation technology of a kind of L-Trp, efficiently solves L-color
The problem that propylhomoserin fermentation middle and late stage Acetic Acid Accumulation too much suppresses fermentation and acid.
For solving above-mentioned technical problem, the technical scheme is that
The fermentation technology of a kind of L-Trp, specifically comprises the following steps that
(1) cottonseed meal adding 2g/L in L-Trp fermentation medium ferments；
(2) sweat is circulated filtration when yield of acetic acid is close or larger than 5g/L, and the fermentation clear liquid after separation is entered
Enter abstraction process；
(3) filter bacterium solution to concentrate through circulation, enter fermentation tank, after adding Dialysis culture base, continue fermentation.
Preferably, the fermentation technology of above-mentioned L-Trp, described L-Trp fermentation medium is glucose 1.5%, phosphorus
Acid hydrogen dipotassium 4.2%, citric acid 0.1%, yeast powder 0.15%, magnesium sulfate 0.1%, manganese sulfate 0.001%, ferrous sulfate
0.005%, remaining is tap water.
Preferably, the fermentation technology of above-mentioned L-Trp, described Dialysis culture base is dipotassium hydrogen phosphate 1.5%, citric acid
0.5%, yeast powder 0.05%, magnesium sulfate 0.4%, remaining is tap water.
Preferably, the fermentation technology of above-mentioned L-Trp, described cottonseed meal is the cottonseed meal of 100 mesh, as thalline
The required nitrogen source of growth, and play filtrating aid function at filter process, improve filtering rate.
Preferably, the fermentation technology of above-mentioned L-Trp, described in be circulated the defecator of filtration and include fermentation tank, gas
Liquid/gas separator, the pump housing, membrane separation plant and storage tank, described fermentation tank discharging opening is connected with the material inlet of gas-liquid separator,
The liquid outlet of described gas-liquid separator is connected with membrane separation plant by the pump housing, described membrane separation plant respectively with storage tank and
Ferment tank connects, and is provided with high temperature high voltage resistant ceramic membrane inside described membrane separation plant, and the gas outlet of described gas-liquid separator passes through
Long tube connects air-filtering membrane.
Preferably, the fermentation technology of above-mentioned L-Trp, the specific operation process of described defecator is as follows:
(1) defecator utilizes 0.1MPa steam to carry out continuous sterilization 1h, utilizes filtrated air to keep malleation after sterilizing；
(2) in fermentation tank, carry out the fermentation of L-Trp, fermentation medium needs add 2g/L cottonseed meal, wait to ferment
Membrane filtration dialysis is carried out close or larger than defecator during 5g/L to acetic acid concentration；
(3) filter process concentrated solution (thalline) directly returns in former fermentation tank, filters bacterium solution volume and is concentrated into Preliminary fermentation
Liquid long-pending 15% time stop filtering, utilize filtrated air that the fermentation liquid in membrane separation plant is pressed into fermentation tank；
(4) after sterilizing and the Dialysis culture base that cools down adds in fermentation tank, addition is according to concentrating 6 times of bacterium solution volume
Adding fermentation tank, proceed fermentation, the stopping fermentation when acetic acid concentration is close to 5g/L in fermentation liquid, repeat the above steps is carried out
Filter dialysis, carry out third time afterwards and ferment (carrying out altogether three fermentations).
Present configuration has the advantages that:
The fermentation technology of above-mentioned L-Trp, has the following technical effect that
1. adding cottonseed meal in fermentation medium, serve not only as the nitrogen source of thalli growth, the cotton seed hulls powder contained is at film
Play the effect of drainage during filtration, make filtering rate increase substantially；
2. fermentation tank is connected with membrane separation device, can open aseptic filtration, the bacterium solution of concentration at any time at limit sweat
Continue fermentation；
3. the present invention carries out filtering dialysis at ferment middle, and the tryptophan, the acetic acid that are contained within by fermentation liquid are filtered to remove, and solves
Inhibitory action thalline produced except tryptophan, acetic acid；
4. the present invention extends the L-Trp fermentation and acid cycle, improves saccharic acid conversion ratio.
Accompanying drawing explanation
Fig. 1 is the structural representation of the fermentation technology of L-Trp of the present invention.
In figure: 1-fermentation tank 2-gas-liquid separator 3-air-filtering membrane 4-gearshift adjustment pump 5-membrane separation plant 6-stores up
Tank
Detailed description of the invention
For further illustrating the present invention, accompanying drawing is now coordinated to be described in detail:
Embodiment 1
The fermentation technology of a kind of L-Trp, specifically comprises the following steps that
(1) as it is shown in figure 1, assembling filter device, described defecator includes 30L fermentation tank 1, gas-liquid separator 2, speed change
Regulation pump 4, membrane separation plant 5 and storage tank 6, described fermentation tank discharging opening and gas-liquid separator (can select whirlwind gas-liquid separation
Device) material inlet be connected, the gas outlet of described gas-liquid separator by long tube connect high-temp. resistant air filter membrane 3, institute
The bottom stating gas-liquid separator is the inverted triangle wimble structure horizontal by 60 ° of angles, and its liquid outlet is positioned at cone bottom, described
Liquid outlet is connected with membrane separation plant by gearshift adjustment pump, and described membrane separation plant connects with storage tank and fermentation tank respectively,
Being provided with high temperature high voltage resistant ceramic membrane inside described membrane separation plant, each parts connect the complete sterilizing that carries out disinfection afterwards.
(2) fermentation medium adds 2g/L cottonseed meal (i.e. adding 2g cottonseed meal in 1L fermentation medium), at warp
Carrying out tryptophan fermentation in fermentation tank after step (1) sterilizing, membrane separation plant utilizes filtrated air to keep barotropic state；Fermentation
It is 31.6g/L that 21h measures L-Trp yield, and yield of acetic acid is 5.2g/L, and now acetic acid produces stronger suppression to L-fermentation,
Opening membrane separation plant to filter, ceramic membrane filter area is 0.12m2。
(3) filter process concentration liquid returns in fermentation tank, and Preliminary fermentation liquid amasss as 20.6L, and filtrate volume is
17.2L, adds 5L sterilized water and continues to filter dialysis, and filtrate volume is 5.1L, and average filtration speed is 185mL/mi n, filters
Utilize filtrated air fermentation liquid press-in fermentation tank in pipeline and in film after end, in fermentation tank, then add 12L dialysis training
Support base, proceed second time fermentation culture.
(4) glucose solution that sweat stream adds 70%, stream sugaring rate controlled adds at fermentation liquid per liter per hour
10-12g glucose, fermentation monitoring acetic acid and L-Trp per hour, when acetic acid concentration is close to 5.0g/L, again carried out
Filter.
(5) second time fermentation 19h, acetic acid concentration is 4.7g/L, and tryptophan yield reaches 29.6g/L, stops second time and sends out
Ferment, then carries out diafiltration.Repeating (3) and (4) step, carry out third time fermentation culture, 18h terminates fermentation.Acid is always produced in fermentation
1.85kg, single tank produces acid ratio batch fermentation and improves 89.9%, and saccharic acid conversion ratio is 18.5%, improves 6.9%.
The fermentation medium of above-mentioned L-Trp is glucose 1.5%, dipotassium hydrogen phosphate 4.2%, citric acid 0.1%, ferment
Female powder 0.15%, magnesium sulfate 0.1%, manganese sulfate 0.001%, ferrous sulfate 0.005%, remaining is tap water；Dialysis culture base
For dipotassium hydrogen phosphate 1.5%, citric acid 0.5%, yeast powder 0.05%, magnesium sulfate 0.4%, remaining is tap water.
Visible, in whole sweat, fermentation L-Trp produces acid more than cycle stretch-out 20h, and acetic acid concentration maintains 5g/
About L, single tank output increased more than 89%, conversion ratio is improved to more than 18.5% by 17.0%；Owing to fermentation medium adding
Adding cottonseed meal, the cotton seed hulls powder not utilized by thalline plays filtrating aid function in filtration, and filtering fermentation liquor speed is by 85mL/mi
N is promoted to 183mL/L.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For Yuan, under the premise without departing from the principles of the invention, it is also possible to make some improvements and modifications, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (6)
1. the fermentation technology of a L-Trp, it is characterised in that: specifically comprise the following steps that
(1) cottonseed meal adding 2g/L in L-Trp fermentation medium ferments；
(2) sweat is circulated filtration when yield of acetic acid is close or larger than 5g/L, and the fermentation clear liquid after separation enters and carries
Take operation；
(3) filter bacterium solution to concentrate through circulation, enter fermentation tank, after adding Dialysis culture base, continue fermentation.
The fermentation technology of L-Trp the most according to claim 1, it is characterised in that: described L-Trp fermentation medium
For glucose 1.5%, dipotassium hydrogen phosphate 4.2%, citric acid 0.1%, yeast powder 0.15%, magnesium sulfate 0.1%, manganese sulfate
0.001%, ferrous sulfate 0.005%, remaining is tap water.
The fermentation technology of L-Trp the most according to claim 1, it is characterised in that: described Dialysis culture base is phosphoric acid hydrogen
Dipotassium 1.5%, citric acid 0.5%, yeast powder 0.05%, magnesium sulfate 0.4%, remaining is tap water.
The fermentation technology of L-Trp the most according to claim 1, it is characterised in that: described cottonseed meal is 100 purposes
Cottonseed meal.
The fermentation technology of L-Trp the most according to claim 1, it is characterised in that be circulated the filtration of filtration described in:
Device includes fermentation tank, gas-liquid separator, the pump housing, membrane separation plant and storage tank, described fermentation tank discharging opening and gas-liquid separator
Material inlet be connected, the liquid outlet of described gas-liquid separator is connected with membrane separation plant by the pump housing, described membrance separation
Equipment connects with storage tank and fermentation tank respectively, is provided with high temperature high voltage resistant ceramic membrane inside described membrane separation plant, and described gas-liquid is divided
Gas outlet from device connects air-filtering membrane by long tube.
The fermentation technology of L-Trp the most according to claim 5, it is characterised in that: the concrete operations of described defecator
Process is as follows:
(1) defecator utilizes 0.1MPa steam to carry out continuous sterilization 1h, utilizes filtrated air to keep malleation after sterilizing；
(2) in fermentation tank, carry out the fermentation of L-Trp, fermentation medium needs add 2g/L cottonseed meal, treat that fermentation is to second
Acid concentration carries out membrane filtration dialysis close or larger than defecator during 5g/L；
(3), in filter process concentrated solution directly returns former fermentation tank, filter bacterium solution volume and be concentrated into what Preliminary fermentation liquid amassed
Stop when 15% filtering, utilize filtrated air that the fermentation liquid in membrane separation plant is pressed into fermentation tank；
(4) after sterilizing and the Dialysis culture base that cools down adds in fermentation tank, addition is according to the 6 times of additions concentrating bacterium solution volume
Fermentation tank, proceeds fermentation, and the stopping fermentation when acetic acid concentration is close to 5g/L in fermentation liquid, repeat the above steps filters
Dialysis, carries out third time afterwards and ferments.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106701535A (en) * | 2017-01-19 | 2017-05-24 | 福建凯立生物制品有限公司 | Zhongshengmycin fermentation continuous feeding production device and production method thereof |
| CN107937452A (en) * | 2017-12-07 | 2018-04-20 | 天津科技大学 | A kind of fermentation manufacturing technique of L tryptophans |
| CN108285913A (en) * | 2017-12-09 | 2018-07-17 | 新疆阜丰生物科技有限公司 | A kind of technique preparing extraction L-Glutamine |
| CN108285914A (en) * | 2017-12-03 | 2018-07-17 | 新疆阜丰生物科技有限公司 | A kind of zymotechnique of L-Trp |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106701535A (en) * | 2017-01-19 | 2017-05-24 | 福建凯立生物制品有限公司 | Zhongshengmycin fermentation continuous feeding production device and production method thereof |
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| CN108285914A (en) * | 2017-12-03 | 2018-07-17 | 新疆阜丰生物科技有限公司 | A kind of zymotechnique of L-Trp |
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| CN108285913A (en) * | 2017-12-09 | 2018-07-17 | 新疆阜丰生物科技有限公司 | A kind of technique preparing extraction L-Glutamine |
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Application publication date: 20161109 |