CN102533891A - Production method of lysine - Google Patents
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- CN102533891A CN102533891A CN2012100094508A CN201210009450A CN102533891A CN 102533891 A CN102533891 A CN 102533891A CN 2012100094508 A CN2012100094508 A CN 2012100094508A CN 201210009450 A CN201210009450 A CN 201210009450A CN 102533891 A CN102533891 A CN 102533891A
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Abstract
The invention discloses a production method of lysine. The method comprises the step of inoculating lysine fermentation strains into lysine fermentation media for fermentation culture under the condition of feeding a carbon source and a nitrogen source. The method is characterized by feeding nutrient salts into fermentation liquor after fermentation culture for 15 hours, wherein the nutrient salts comprise potassium chloride, magnesium sulfate and manganese sulfate. The production method has the following advantages: the content of final lysine, the acid supply quantity per tank and the conversion rate can be improved, namely improving the yield of lysine, thus improving the economic benefits.
Description
Technical field
The present invention relates to a kind of working method of Methionin.
Background technology
At present China's fermenting lysine all adopts the method for batch fermentation, and fermentation adopts every jar to drop into fermention medium and insert certain fermenting lysine bacterial classification according to certain ratio, adds carbon source at stream and adds under the condition of nitrogenous source with stream, carries out fermentation culture.Along with the carrying out of fermentation, Methionin acidity improves gradually, and along with the stream of carbon source and nitrogenous source add and fermenting process in blowing; Other nutrient concentrations in the fermentor tank beyond carbon, the nitrogen reduce gradually, and the Methionin bacterial classification is old and feeble too early, self-dissolving, and metabolic capacity descends; Fermentation and acid speed is on a declining curve, finally slowly puts jar because of fermentation and acid speed, causes the production capacity that has limited Methionin; Cost is higher, and economic benefit is lower.
Summary of the invention
The objective of the invention is in order to improve the production capacity of Methionin, thereby increase economic efficiency, a kind of working method of new Methionin is provided.
Contriver of the present invention is unexpected under study for action to be found; Fermentation culture after 15 hours in fermented liquid stream add the nutritive salt that comprises Repone K, sal epsom and manganous sulfate; Can greatly improve terminal point lysine content, single jar of sour the measuring and transformation efficiency of confession, promptly improve the Methionin production capacity, thereby increase economic efficiency.
Therefore, to achieve these goals, the invention provides a kind of working method of Methionin; Said method comprises in the fermenting lysine bacterial classification access fermenting lysine substratum, adds carbon source at stream and adds under the condition of nitrogenous source with stream, carries out fermentation culture; It is characterized in that; After 15 hours, in fermented liquid, flow Ensure Liquid salt in fermentation culture, said nutritive salt comprises Repone K, sal epsom and manganous sulfate.
Preferably, finish in preceding 6 hours to fermentation culture after 15 hours in fermentation culture, stream adds said nutritive salt in fermented liquid.
The working method of Methionin provided by the invention can improve terminal point lysine content, single jar of sour the measuring and transformation efficiency of confession, promptly improves the Methionin production capacity, thereby increases economic efficiency.
Other features and advantages of the present invention will partly specify in embodiment subsequently.
Embodiment
Following specific embodiments of the invention is elaborated.Should be understood that embodiment described herein only is used for explanation and explains the present invention, is not limited to the present invention.
The invention provides a kind of working method of Methionin; This method comprises in the fermenting lysine bacterial classification access fermenting lysine substratum; Add carbon source at stream and add under the condition of nitrogenous source, carry out fermentation culture, in fermentation culture after 15 hours with stream; Stream Ensure Liquid salt in fermented liquid, nutritive salt comprises Repone K, sal epsom and manganous sulfate.
Among the present invention; Fermention medium is for well known to a person skilled in the art notion; Refer to the nutriment of required confession microorganism growth of microbial fermentation and the manual work preparation of keeping usefulness, generally all contain glucide, nitrogenous substances, inorganic salt (comprising trace element) and VITAMINs and water etc.Fermented liquid refers to an access the liquid nutrient medium (this liquid nutrient medium also is an alleged fermention medium among the present invention) of microorganism strains, products therefrom after cultivation after a while also for well known to a person skilled in the art notion.
According to the present invention, although after 15 hours, in fermented liquid, flow Ensure Liquid salt in fermentation culture; Nutritive salt comprises Repone K, sal epsom and manganous sulfate, can realize the object of the invention, promptly improves terminal point lysine content, single jar of sour the measuring and transformation efficiency of confession; But in order further to reduce cost, and do not influence the raising of Methionin production capacity, under the preferable case; Finish in preceding 6 hours to fermentation culture after 15 hours in fermentation culture, stream adds the nutritive salt that comprises Repone K, sal epsom and manganous sulfate in fermented liquid.
Among the present invention, to add carbon source be benchmark with the fermention medium that stream adds before the nitrogenous source to inoculate back and stream, and the inoculum size of fermenting lysine bacterial classification is 12-18 volume %.What those skilled in the art should understand that is; The fermenting lysine bacterial classification in being seeded to fermention medium before; Adopt ordinary method with fermenting lysine bacterial classification process seed tank culture; Cultivation degree in seed tank culture can be observed producing the Methionin microbial growth through sampling sediments microscope inspection, OD (optical density) pH-value determination pH; When observe through aforesaid method thalli morphology normal, measure OD value and reach 0.75 and stop cultivation when above, the seed liquor in this moment seeding tank is called mature seed liquid.And then with in the mature seed liquid access fermention medium.Therefore, among the present invention, the inoculum size of fermenting lysine bacterial classification is 12-18 volume %, refers to the volume that inserts the mature seed liquid in the fermention medium and accounts for the long-pending 12-18% of access mature seed liquid post-fermentation and culture matrix.
Seed tank culture can adopt first class seed pot to cultivate also can adopt the cultivation of secondary seed jar, and the first class seed pot cultivation is about to the fermenting lysine bacterial classification and in a seeding tank, cultivates required cultivation degree always; The secondary seed jar changes another seeding tank continuation cultivation over to after cultivating and promptly earlier the fermenting lysine bacterial classification being cultivated for some time in a seeding tank again, cultivates required cultivation degree.The secondary seed jar is cultivated in the not concrete qualification of the incubation time of each seeding tank, as long as finally can cultivate required cultivation degree.For easy to operate, seed tank culture of the present invention preferably adopts first class seed pot to cultivate.
Among the present invention; Composition for the seed tank culture base does not have particular requirement; This area seed tank culture base commonly used can be adopted, for example, preparation seed tank culture bases such as starchy material saccharification clear liquid, steeping water, potassium hydrogenphosphate, sal epsom, ammonium sulfate, Threonine and methionine(Met) can be used.According to the present invention, the consumption of each raw material can in very large range change in every liter of seed tank culture base, under the preferable case; In every liter of seed tank culture base; The consumption of starchy material saccharification clear liquid can restrain for 30-40, and the consumption of steeping water (dry weight is 20-50 weight %) can restrain for 70-90, and the consumption of potassium hydrogenphosphate can restrain for 0.5-1.5; The consumption of sal epsom can restrain for 0.4-1.1; The consumption of ammonium sulfate can restrain for 5-15, and the consumption of Threonine can restrain for 0.1-0.6, and the consumption of methionine(Met) can restrain for 0.1-0.3.
In order further to improve the production capacity of Methionin; The amount that stream adds Repone K makes preferably that the concentration of Repone K is controlled at the 2-4 grams per liter in the fermented liquid; The amount that stream adds sal epsom makes preferably that the concentration of sal epsom is controlled at the 1-3 grams per liter in the fermented liquid, and the amount that stream adds manganous sulfate makes preferably that the concentration of manganous sulfate is controlled at the 0.02-0.06 grams per liter in the fermented liquid.The concentration and the flow velocity of the nutritive salt that adds for stream do not have particular requirement, and the concentration of each nutritive salt is controlled in the above-mentioned scope in the fermented liquid as long as make.
Among the present invention, the amount that stream adds carbon source makes that the concentration of reducing sugar is controlled at the 5-10 grams per liter in the fermented liquid, and the amount that stream adds nitrogenous source makes that the concentration of nitrogen is controlled at the 0.35-0.8 grams per liter in the fermented liquid." amount that stream adds carbon source makes that the concentration of reducing sugar is controlled at the 5-10 grams per liter in the fermented liquid; the amount that stream adds nitrogenous source makes that the concentration of nitrogen is controlled at the 0.35-0.8 grams per liter in the fermented liquid " is meant that adding speed that carbon source and stream adds nitrogenous source through controlling flow makes in whole fermentation culture process that the concentration of reducing sugar maintains the 5-10 grams per liter in the fermented liquid here, makes that the concentration of nitrogen maintains the 0.35-0.8 grams per liter in the fermented liquid.
Contriver of the present invention finds in experiment, add for the stream of carbon source and nitrogenous source, and the stream of nutritive salt adds, and it is good that even flow adds the ferment effect that adds than intermittent flow, and therefore, the stream among the present invention adds and is preferably even flow and adds.
Among the present invention, fermentation culture is carried out in fermentor tank, in order effectively to utilize the production capacity of fermentor tank; Inoculation back and stream add the 40-60% that volume that carbon source and stream adds the substratum in the fermentor tank before the nitrogenous source is preferably the fermentor tank volume; Along with stream adds carbon source and stream adds nitrogenous source, and fermentation culture is after 15 hours, stream Ensure Liquid salt in fermented liquid; The volume of the substratum in the fermentor tank increases gradually; In order to guarantee the air capacity in the fermentor tank, be preferably and flow blowing when adding carbon source and flowing the 70-80% that adds nitrogenous source and stream Ensure Liquid salt to fermentor tank volume, in order to guarantee the bacterial classification quantity in the blowing secondary fermentation jar; Do not influence the fermentation culture in the blowing secondary fermentation jar, the blowing volume is preferably the 5-10% of culture volume in the blowing prefermentor.
In the prior art, the fermenting lysine bacterial classification is inserted in the fermenting lysine substratum, add carbon source at stream and add under the condition of nitrogenous source with stream; Carry out fermentation culture, along with the carrying out of fermentation, Methionin acidity improves gradually; Nutritive substance reduces gradually, and fermentation and acid speed is on a declining curve, after nutritive substance is reduced to a certain degree; Fermentation and acid speed is reduced to and to a certain degree promptly is judged as fermentation termination; Reach fermentation termination and promptly put jar, put jar and be meant the substratum in the fermentor tank is all emitted from fermentor tank, promptly stop fermentation.Generally put jar after fermentation culture 40-50 hour in the prior art.The present invention is because in fermentation culture after 15 hours, and stream Ensure Liquid salt in fermented liquid, but therefore time of proper extension fermentation culture are preferably put jar after fermentation culture 42-54 hour.
It will be understood by those skilled in the art that in order to obtain the higher Methionin product of purity the inventive method also comprises from blowing or the solution put jar extracts Methionin.Method for extracting Methionin does not have particular requirement; Can adopt this area the whole bag of tricks commonly used, for example, adopt continuous ionic exchange separating and extracting method; In blowing or the solution put jar, adding a large amount of vitriol oils transfers pH to 2.0-3.0 to carry out acidifying; Remove thalline through metallic membrane or ceramic membrane filter after the acidifying, obtaining Methionin membrane filtration liquid is the Methionin clear liquid, or the lysine fermentation liquor after the acidifying is obtained the Methionin clear liquid after flocculation filtration removes thalline; Except that adopting strongly acidic cation-exchange, the Methionin clear liquid behind the thalline adsorbs exchange; Wash-out is carried out with weak ammonia in the saturated back of resin absorption, and the Methionin that elutes obtains the lysine hydrochloride finished product through concentrated, salt acid for adjusting pH value, crystallization, solid-liquid separation, oven dry; Perhaps in blowing or the solution put jar, add acid, make the lysine fermentation liquor acidifying, remove thalline through method such as filtering or centrifugal.Add calcium hydroxide in the Methionin clear liquid after removing thalline and regulate the pH value to 8.0-11.5; Make impurity such as salt in the Methionin clear liquid, colloid generate insolubles; After solid-liquid separation, obtain lysine solution, the content that lysine solution is concentrated into Methionin in every milliliter of lysine solution is 0.6-0.8g, can obtain highly purified lysine solution through filtering again; Through concentrated, salt acid for adjusting pH value, crystallization, solid-liquid separation, oven dry, obtain the lysine hydrochloride finished product then.
Among the present invention, the condition of fermentation culture is not had particular requirement, can adopt this area condition commonly used; For example; Temperature is 35-38 ℃, and pressure is 0.05-0.1MPa, and the pH value is 6.7-7.0; Air flow be 0.5-1.2 cubic metres of air/cubic meter substratum/minute, air flow be preferably the substratum of 0.7-0.9 cubic metres of air/cubic meter/minute.
Among the present invention, the kind of fermenting lysine bacterial classification is not had particular requirement, can adopt this area bacterial classification commonly used, be preferably at least a in Corynebacterium glutamicum, intestinal bacteria and the brevibacterium flavum.
Among the present invention, carbon source is preferably starchy material saccharification clear liquid.
Among the present invention, starchy material saccharification clear liquid both can adopt the preparation of dry method sugar refining technology, also can adopt the preparation of wet method sugar refining technology.Simple from technology, facility investment is few, the lower aspect of production cost is considered, preferably through the preparation of dry method sugar refining technology.The dry method sugar refining technology is meant that starchy material directly carries out fragmentation and enzymolysis without soaking.
The dry method sugar refining technology can comprise: starchy material is pulverized, and the product after starchy material is pulverized is sized mixing, and adding glycase carries out the hydrolysis first time to starch; To the first time hydrolysate carry out solid-liquid separation, and in the liquid phase component that obtains, add saccharifying enzyme and carry out the hydrolysis second time, obtain starchy material saccharification clear liquid.Preferably, pulverize and to make percent of pass that starchy material crosses 30 mesh sieves greater than 75%, the percent of pass of more preferably crossing 30 mesh sieves is 100%.The method of sizing mixing is well known to those skilled in the art, but preferably, and the method for sizing mixing can comprise that the product after starchy material pulverized is added to the water and mix that the add-on of water makes the degree Beaume of the slurries that obtain be 9-17B é °.Term " degree Beaume " is a kind of method of expression strength of solution, is to detect the number of degrees that solution obtains through Beaum.
According to the present invention, in the hydrolysis first time, with the dry weight basis of the product after every gram pulverizing; Diastatic consumption can be the 10-30 enzyme activity unit; The temperature of enzymolysis can be 88-92 ℃, and the time of enzymolysis can be 90-120 minute, and the pH value of enzymolysis can be 5.5-6.0.The condition of solid-liquid separation does not have special qualification, and preferably, it is 19-22 weight %, more preferably 20-21 weight % that the condition of solid-liquid separation makes the solid content in the liquid phase component that obtains.
According to the present invention, in the hydrolysis second time, in every gram liquid phase component, the consumption of saccharifying enzyme can be the 110-130 enzyme activity unit, and the temperature of enzymolysis can be 55-65 ℃, and the time of enzymolysis can be 420-600 minute, and the pH value of enzymolysis can be 4.0-4.5.
Being defined as of enzyme activity unit of the present invention: be 6.0 in the pH value, temperature is that 1 minute is converted into the required enzyme amount of reducing sugar with 1 milligram of starch is an enzyme activity unit under 70 ℃ the condition.
Glycase is meant the general name of class of enzymes that can the starch-splitting glycosidic link, and said glycase generally comprises AMS, beta-amylase.
AMS is claimed starch 1 again, the 4-dextrinase, and it can cut the inner α-1 of starch chain at random, brokenly, and the 4-glycosidic link is hydrolyzed to starch SANMALT-S, contains the oligosaccharides of 6 glucose units and has the oligosaccharides of side chain.The mikrobe that produces this enzyme mainly has Bacillus subtilus, black mold, aspergillus oryzae and head mold.
Beta-amylase is claimed starch 1 again, and 4-maltoside enzyme can cut 1 from the starch molecule non reducing end, and the 4-glycosidic link generates SANMALT-S.The product that this enzyme acts on starch is SANMALT-S and limit dextrin.This enzyme is mainly produced by aspergillus, head mold and endomyces.
According to the present invention, preferably use AMS.
According to the present invention, saccharifying enzyme is preferably α-1,4-glucose hydrolysis enzyme.
According to the present invention, starchy material can be the various raw materials that contain starch that can be used for enzymolysis, fermentative prepn Methionin well known in the art, for example, can be selected from corn, potato class (like cassava) and the wheat one or more.
Among the present invention, the kind of nitrogenous source is conventionally known to one of skill in the art, for example, can be ammonium salt.When nitrogenous source was ammonium salt, the concentration of nitrogen was represented with the concentration of ammonium radical ion in the substratum, and the concentration of nitrogen is controlled at the 0.35-0.8 grams per liter, and then the concentration of ammonium radical ion is controlled at the 0.5-1.0 grams per liter.
Among the present invention; The composition of fermention medium does not have particular requirement; Can adopt this area fermenting lysine substratum commonly used; For example, can use preparation fermention mediums such as starchy material saccharification clear liquid, molasses, steeping water, ammonium sulfate, potassium hydrogenphosphate, sal epsom, Threonine, methionine(Met) and L-glutamic acid.According to the present invention, the consumption of each raw material can in very large range change in every liter of fermention medium, under the preferable case; In every liter of fermention medium, the consumption of starchy material saccharification clear liquid can restrain for 40-60, and the consumption of molasses can restrain for 30-50; The consumption of steeping water (dry weight is 20-50 weight %) can restrain for 20-40, and the consumption of ammonium sulfate can restrain for 20-40, and the consumption of potassium hydrogenphosphate can restrain for 0.5-1.5; The consumption of sal epsom can restrain for 0.4-0.6; The consumption of Threonine can restrain for 0.1-0.3, and the consumption of methionine(Met) can restrain for 0.1-0.3, and the consumption of L-glutamic acid can restrain for 0.2-0.4.
In addition, it will be understood by those skilled in the art that the bacterial classification that the access seeding tank is cultivated is the bacterial classification behind the laggard capable multiplication culture of overactivation.Activation and multiplication culture are the common practise of this area, repeat no more at this.
More than describe preferred implementation of the present invention in detail; But the present invention is not limited to the detail in the above-mentioned embodiment, in technical conceive scope of the present invention; Can carry out multiple simple variant to technical scheme of the present invention, these simple variant all belong to protection scope of the present invention.
Need to prove in addition; Each concrete technical characterictic described in above-mentioned embodiment under reconcilable situation, can make up through any suitable manner; For fear of unnecessary repetition, the present invention is to the explanation no longer separately of various possible array modes.
In addition, also can carry out arbitrary combination between the various embodiment of the present invention, as long as it is without prejudice to thought of the present invention, it should be regarded as the disclosed content of the present invention equally.
Embodiment
Following embodiment will be further described the present invention, but therefore not limit the present invention.
In following embodiment and Comparative Examples:
OD pH-value determination pH: the fermented liquid of sampling is carried out 26 times of dilutions; Adopt the 722N visible spectrophotometer; Under wavelength 562 nanometer visible lights, measure light absorption value; Be the OD value,, produce the Methionin microbial numbers in the numerical value that the calculates reflection unit volume fermented liquid the TV of the OD value that obtains * extension rate ÷ fermented liquid.
Measure the concentration of reducing sugar in the fermented liquid according to the method for GB/T5009.7-2008.
Measure the concentration of ammonium radical ion in the fermented liquid according to the method for GB3595-83.
Measure the concentration of Repone K in the fermented liquid according to the method for GB3595-1996.
Measure the concentration of sal epsom in the fermented liquid according to the method for GB/T671-1998.
Measure the concentration of manganous sulfate in the fermented liquid according to the method for GB/T15899-1995.
According to the lysine concentration (in lysine hydrochloride) in the GB10794-89 standard detection substratum.
Single jar supplies acid amount=(lysine concentration of putting jar * put tank volume+middle blowing lysine concentration * middle blowing volume).
Transformation efficiency (%)=single jar supplies weight * 100% of acid amount/total reducing sugar, and wherein the weight of total reducing sugar comprises that seeding tank uses sugar weight with sugar weight and fermentor tank.
Embodiment 1
Present embodiment is used to explain the working method of Methionin provided by the invention.
(1) the 100 weight part corns that will gather in the crops are pulverized corn particle through mechanical workout, and the percent of pass that makes Semen Maydis powder cross 30 mesh sieves is 80%.
(2) product after will pulverizing adds water and sizes mixing to 12B é °, with respect to the dry weight of every gram crushed products, adds the glycase (Novozymes Company, AMS) of 20 enzyme activity units, is enzymolysis 100 minutes under 5.5 the condition at 90 ℃, pH, obtains enzymolysis product.Wherein, enzymolysis product through carrying out press filtration with the fluid pressure type plate-and-frame filter press, is isolated enzymolysis clear liquid (solid content is 20 weight %); The saccharifying enzyme (α-1,4-glucose hydrolysis enzyme, Novozymes Company) that adds 115 enzyme activity units afterwards is enzymolysis 420 minutes under 4.5 the condition at 60 ℃, pH, obtains starchy material saccharification clear liquid.
(3) the starchy material saccharification clear liquid preparation seed tank culture base that uses step (2) to obtain specifically consists of: with respect to every liter substratum, the consumption of starchy material saccharification clear liquid is 35 grams; The consumption of steeping water (dry weight is 35 weight %) is 80 grams; The consumption of potassium hydrogenphosphate is 1.0 grams, and the consumption of sal epsom is 0.5 gram, and the consumption of ammonium sulfate is 10 grams; The consumption of Threonine is 0.2 gram, and the consumption of methionine(Met) is 0.2 gram.Substratum is heated to 121 ℃ of sterilizations, keeps being cooled to 37 ℃ and keep constant after 20 minutes.Open and stir, the adjusting tank pressure is 0.1MPa, feeds sterile air according to ventilation and 1: 0.5 volume ratio of substratum, regulates pH to 6.8 and keeps constant with ammoniacal liquor.Then brevibacterium flavum bacterial classification (bacterial strain original seed FB42 is available from Southern Yangtze University) activation is inserted in the seeding tank with the propagation back and cultivate; Every microscopies and measure the OD value of taking a sample at a distance from 120 minutes in the culturing process; Stop cultivation when normal and OD value reaches 0.8 when the microscopy thalli morphology, obtain mature seed liquid.
(4) the starchy material saccharification clear liquid preparation fermention medium that uses step (2) to obtain specifically consists of: with respect to every liter of fermention medium, the consumption of starchy material saccharification clear liquid is 50 grams; The consumption of molasses (Xinjiang, the place of production) is 40 grams; The consumption of steeping water (dry weight is 35 weight %) is 30 grams, and the consumption of ammonium sulfate is 30 grams, and the consumption of potassium hydrogenphosphate is 1.0 grams; The consumption of sal epsom is 0.5 gram; The consumption of Threonine is 0.2 gram, and the consumption of methionine(Met) is 0.2 gram, and the consumption of L-glutamic acid is 0.3 gram.Substratum is heated to 121 ℃ of sterilizations and is cooled to 37 ℃ and keep constant after 30 minutes, regulates pH to 6.9 with ammoniacal liquor.
(5) fermention medium that step (4) prepares of in fermentor tank, packing into, the fermention medium volume is 50% of a fermentor tank volume.Using the mature seed liquid of step (3) gained, carry out fermentation culture in the substratum of access fermentor tank, is benchmark with postvaccinal fermention medium, and the inoculum size of the mature seed liquid of step (3) gained is 15 volume %.Inoculation back even flow adds starchy material saccharification clear liquid and the ammonium sulfate that step (2) obtains; The amount that stream adds the starchy material saccharification clear liquid that step (2) obtains makes that the concentration of reducing sugar is controlled at the 6-8 grams per liter in the fermented liquid; The amount that stream adds ammonium sulfate makes the concentration of ammonium radical ion in the fermented liquid be controlled at the 0.6-0.8 grams per liter; Tank pressure is controlled to be 0.1MPa, and leavening temperature is controlled to be 37 ℃, air flow be 0.7 cubic metres of air/cubic meter substratum/minute; And regulate pH with liquefied ammonia and maintain 6.9 and carry out fermentation culture; Finish in preceding 6 hours to fermentation culture after 15 hours in fermentation culture, stream adds Repone K, sal epsom and manganous sulfate in fermented liquid, and the amount that stream adds Repone K makes that the concentration of Repone K is controlled at the 2-4 grams per liter in the fermented liquid; The amount that stream adds sal epsom makes that the concentration of sal epsom is controlled at the 1-3 grams per liter in the fermented liquid, and the amount that stream adds manganous sulfate makes that the concentration of manganous sulfate is controlled at the 0.02-0.06 grams per liter in the fermented liquid.
Stream adds starchy material saccharification clear liquid and the ammonium sulfate that step (2) obtains, and stream adds 75% o'clock blowing of Repone K, sal epsom and manganous sulfate to fermentor tank volume, the blowing volume be in the blowing prefermentor culture volume 8%.Fermentation culture was put jar after 48 hours, measured the lysine concentration of the lysine concentration put jar (be the terminal point lysine content, down with) and middle blowing, calculated single jar and supplied sour measure and transformation efficiency is seen table 1.
Embodiment 2
Present embodiment is used to explain the working method of Methionin provided by the invention.
The preparation method of starchy material saccharification clear liquid, seed tank culture based formulas, seeding tank mature seed liquid cultural method, fermentor cultivation based formulas are all identical with embodiment 1.
Culture volume in the fermentor tank is 40% of a fermentor tank volume.Carrying out fermentation culture in the substratum with mature seed liquid access fermentor tank, is benchmark with postvaccinal fermention medium, and the inoculum size of seed liquor is 12 volume %.Inoculation back even flow adds starchy material saccharification clear liquid and the ammonium sulfate that makes; The amount that stream adds the starchy material saccharification clear liquid that makes makes that the concentration of reducing sugar is controlled at the 5-7 grams per liter in the fermented liquid; The amount that stream adds ammonium sulfate makes the concentration of ammonium radical ion in the fermented liquid be controlled at the 0.5-0.7 grams per liter; Tank pressure is controlled to be 0.08MPa, and leavening temperature is controlled to be 35 ℃, air flow be 0.8 cubic metres of air/cubic meter substratum/minute; And regulate pH with liquefied ammonia and maintain 6.7 and carry out fermentation culture; Finish in preceding 6 hours to fermentation culture after 15 hours in fermentation culture, stream adds Repone K, sal epsom and manganous sulfate in fermented liquid, and the amount that stream adds Repone K makes that the concentration of Repone K is controlled at the 2-4 grams per liter in the fermented liquid; The amount that stream adds sal epsom makes that the concentration of sal epsom is controlled at the 1-3 grams per liter in the fermented liquid, and the amount that stream adds manganous sulfate makes that the concentration of manganous sulfate is controlled at the 0.02-0.06 grams per liter in the fermented liquid.
Stream adds starchy material saccharification clear liquid and the ammonium sulfate that makes, and stream adds 70% o'clock blowing of Repone K, sal epsom and manganous sulfate to fermentor tank volume, the blowing volume be in the blowing prefermentor culture volume 5%.Fermentation culture was put jar after 42 hours, and mensuration is put the lysine concentration of jar and the lysine concentration of middle blowing, calculated single jar and supplied acid amount and transformation efficiency to see table 1.
Embodiment 3
Present embodiment is used to explain the working method of Methionin provided by the invention.
The preparation method of starchy material saccharification clear liquid, seed tank culture based formulas, seeding tank mature seed liquid cultural method, fermentor cultivation based formulas are all identical with embodiment 1.
Culture volume in the fermentor tank is 60% of a fermentor tank volume.Carrying out fermentation culture in the substratum with mature seed liquid access fermentor tank, is benchmark with postvaccinal fermention medium, and the inoculum size of seed liquor is 18 volume %.Inoculation back even flow adds starchy material saccharification clear liquid and the ammonium sulfate that makes; The amount that stream adds the starchy material saccharification clear liquid that makes makes that the concentration of reducing sugar is controlled at the 8-10 grams per liter in the fermented liquid; The amount that stream adds ammonium sulfate makes the concentration of ammonium radical ion in the fermented liquid be controlled at the 0.8-1.0 grams per liter; Tank pressure is controlled to be 0.05MPa, and leavening temperature is controlled to be 38 ℃, air flow be 0.9 cubic metres of air/cubic meter substratum/minute; And regulate pH with liquefied ammonia and maintain 7.0 and carry out fermentation culture; Finish in preceding 6 hours to fermentation culture after 15 hours in fermentation culture, stream adds Repone K, sal epsom and manganous sulfate in fermented liquid, and the amount that stream adds Repone K makes that the concentration of Repone K is controlled at the 2-4 grams per liter in the fermented liquid; The amount that stream adds sal epsom makes that the concentration of sal epsom is controlled at the 1-3 grams per liter in the fermented liquid, and the amount that stream adds manganous sulfate makes that the concentration of manganous sulfate is controlled at the 0.02-0.06 grams per liter in the fermented liquid.
Stream adds starchy material saccharification clear liquid and the ammonium sulfate that makes, and stream adds 80% o'clock blowing of Repone K, sal epsom and manganous sulfate to fermentor tank volume, the blowing volume be in the blowing prefermentor culture volume 10%.Fermentation culture was put jar after 54 hours, and mensuration is put the lysine concentration of jar and the lysine concentration of middle blowing, calculated single jar and supplied acid amount and transformation efficiency to see table 1.
Comparative Examples 1
Produce Methionin according to the method for embodiment 1, different is, in the fermentation culture process in fermented liquid stream add Repone K, sal epsom and manganous sulfate.Fermentation culture was put jar after 48 hours, and mensuration is put the lysine concentration of jar and the lysine concentration of middle blowing, calculated single jar and supplied acid amount and transformation efficiency to see table 1.
Table 1
| Terminal point lysine content (g/100mL) | Single jar supplies acid amount (t) | Transformation efficiency (%) | |
| Embodiment 1 | 18.35 | 52.62 | 63.25 |
| Embodiment 2 | 18.20 | 50.04 | 60.09 |
| Embodiment 3 | 18.12 | 53.51 | 62.35 |
| Comparative Examples 1 | 17.34 | 48.03 | 59.95 |
From table 1, can find out, adopt the inventive method fermentative prodn Methionin, can obviously improve terminal point lysine content, single jar of sour the measuring and transformation efficiency of confession.
The working method of Methionin provided by the invention; Through in fermentation culture after 15 hours; Stream adds the nutritive salt that comprises Repone K, sal epsom and manganous sulfate in fermented liquid; Improved terminal point lysine content, single jar of sour the measuring and transformation efficiency of confession, promptly improved the Methionin production capacity, thereby improved economic benefit.
Claims (13)
1. the working method of a Methionin; Said method comprises in the fermenting lysine bacterial classification access fermenting lysine substratum, adds carbon source at stream and adds under the condition of nitrogenous source with stream, carries out fermentation culture; It is characterized in that; After 15 hours, in fermented liquid, flow Ensure Liquid salt in fermentation culture, said nutritive salt comprises Repone K, sal epsom and manganous sulfate.
2. method according to claim 1 wherein, finishes in preceding 6 hours to fermentation culture after 15 hours in fermentation culture, and stream adds said nutritive salt in fermented liquid.
3. method according to claim 1 and 2; Wherein, The amount that stream adds Repone K makes that the concentration of Repone K is controlled at the 2-4 grams per liter in the fermented liquid; The amount that stream adds sal epsom makes that the concentration of sal epsom is controlled at the 1-3 grams per liter in the fermented liquid, and the amount that stream adds manganous sulfate makes that the concentration of manganous sulfate is controlled at the 0.02-0.06 grams per liter in the fermented liquid.
4. according to any described method among the claim 1-3, wherein, to add the fermention medium that carbon source and stream adds before the nitrogenous source be benchmark to inoculate back and stream, and the inoculum size of said fermenting lysine bacterial classification is 12-18 volume %.
5. method according to claim 4, wherein, the amount that said stream adds carbon source makes that the concentration of reducing sugar is controlled at the 5-10 grams per liter in the fermented liquid, and the amount that said stream adds nitrogenous source makes that the concentration of nitrogen is controlled at the 0.35-0.8 grams per liter in the fermented liquid.
6. according to any described method among the claim 1-5, wherein, said stream adds for even flow and adds.
7. according to any described method among the claim 1-6; Wherein, Said fermentation culture is carried out in fermentor tank; It is the 40-60% of fermentor tank volume that inoculation back and stream add the volume that carbon source and stream adds the substratum in the fermentor tank before the nitrogenous source, blowing when stream adds carbon source and nitrogenous source and flows the 70-80% of Ensure Liquid salt to fermentor tank volume, and the blowing volume is the 5-10% of culture volume in the blowing prefermentor.
8. method according to claim 7 wherein, is put jar after fermentation culture 42-54 hour.
9. according to claim 7 or 8 described methods, wherein, said method also comprises from the solution that said blowing or said is put jar extracts Methionin.
10. according to any described method among the claim 1-9, wherein, the condition of said fermentation culture is: temperature is 35-38 ℃, and pressure is 0.05-0.1MPa, and the pH value is 6.7-7.0, air flow be 0.5-1.2 cubic metres of air/cubic meter substratum/minute.
11. according to any described method among the claim 1-10, wherein, said fermenting lysine bacterial classification is at least a in Corynebacterium glutamicum, intestinal bacteria and the brevibacterium flavum.
12. according to any described method among the claim 1-11, wherein, said carbon source is a starchy material saccharification clear liquid.
13. according to any described method among the claim 1-12, wherein, said nitrogenous source is an ammonium salt.
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| CN110872606A (en) * | 2019-09-02 | 2020-03-10 | 黑龙江成福食品集团有限公司 | Preparation method of L-lysine sulfate and by-product thereof |
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| CN104232702B (en) * | 2014-07-28 | 2020-02-14 | 中粮生物科技股份有限公司 | Production method of lysine |
| CN106148443A (en) * | 2015-04-02 | 2016-11-23 | 中粮生物化学（安徽）股份有限公司 | A kind of method producing lysine |
| 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 |
| CN110872606A (en) * | 2019-09-02 | 2020-03-10 | 黑龙江成福食品集团有限公司 | Preparation method of L-lysine sulfate and by-product thereof |
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