CN106635934A - Thermophilic type lactobacillus and method for soaking corns by manually adding thermophilic type lactobacillus - Google Patents
Thermophilic type lactobacillus and method for soaking corns by manually adding thermophilic type lactobacillus Download PDFInfo
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Abstract
The invention relates to thermophilic type lactobacillus and a method for soaking corns by manually adding the thermophilic type lactobacillus. A name of the thermophilic type lactobacillus is C15, a systematic name of the thermophilic type lactobacillus is bacillus (Bacillus sp.), a preservation serial number of the thermophilic type lactobacillus is CGMCC No. 13471, a preservation date is December 19, 2016, and a preservation unit is China general microbiological culture collection center with an address of No. 3, No.1 courtyard, Beichen west road, Chaoyang District, Beijing. By means of the method of adding the certain amount of thermophilic type lactobacillus into corn soaking solution, a corn kernel soaking effect is enhanced, corn soaking time is shortened, the quality of the corn soaking solution is improved, the acid yield and the conversion rate of thermo-sensitive type glutamic acid fermentation with high stability are guaranteed, a good production condition is provided for monosodium glutamate production, and the monosodium glutamate production cost is reduced.
Description
Technical field
The invention belongs to bio-fermentation engineering field, is related to the production of corn soaking, especially a kind of thermophilic type is newborn
Acidfast bacilli and manually add the corn soaking method of the thermophilic type lactobacilluss.
Background technology
The preparation of corn starch is divided to two kinds of dry and wet.So-called dry method refers to the method by grinding screening selecting crude drugs with winnower, separates
Plumule and fiber, and obtain low-fat Semen Maydis powder.Wet method refers to " one leaching two mill three points ", will Semen Maydiss warm water soaking, corase grind,
Fine grinding, separated plumule, cellulose and protein, and obtain high-purity starch.To obtain highly purified corn starch, typically all adopt
Carried out with closed wet processing.
Semen Maydis pulp contains abundant aminoacid and vitamin, promotees as cheap organic nitrogen source in glutamic acid fermentation production
Enter thalli growth, affect the acid production rate and saccharic acid conversion ratio of fermentation.In actual production, during due to by Semen Maydiss quality and immersion
Between the short impact for causing Semen Maydis pulp quality unstable, the production of temperature sensitive type glutamic acid fermentation seriously restricted.
By retrieval, following two patent publication us related to present patent application are found：
1st, a kind of lactobacillus inoculum method (publication number 102942632B) for improving corn soaking effect, is related to a kind of use
In the lactobacillus inoculum method for improving corn soaking effect, old acid in circulation impregnating autoclave group is set to enter the newly-increased old acid by pressure reduction
7～9 hours are placed in tank, the old acid in the newly-increased old acid tank, and temperature 50 C, lactic acid content reaches 2.2%～2.6%；It is right
SO2The old acid being worth after the culture in the newly-increased old acid tank of the steeping tank addition of 100ppm～200ppm, according to impregnation liquid：Culture
Old acid=30 afterwards:1 part by weight is added, and keeps liquid level of the old acid tank constant, has obtained improving breast in soak in big production
The acid content best approach, improves immersion effect and starch recoveries.
2nd, the corn starch soaking prodn. process of adding vegetable protease (patent No.：200610016774), it is related to agricultural byproducts intensive processing,
More particularly to corn starch soaking prodn. process of adding vegetable protease.It overcomes traditional Corn Starch Wet production technology and there is leaching
The deficiencies such as bubble overlong time, water and big, the sulfur dioxide pollution environment of energy resource consumption.Its production craft step is：The first step, selection-breeding
Suitable corn steep liquor lactobacillus inoculation, will be added in corn steep liquor, to shorten effective lactic acid concn after its fermentation culture
The time for reaching, soften the imbibition of corn kernel endosperm, cell wall is destroyed；Second step, the Semen Maydiss of first leaching are crushed
It is that lactic acid and penetrating for protease create more preferable condition into 4-6 lobes, separated plumule；3rd step, in broken corn soaking
Plant rennet is added in liquid, starch is discharged with abundant protein degradation substrate, further shorten soak time.
By contrast, there are the different of essence in present patent application from above-mentioned patent publication us.
The content of the invention
It is an object of the invention to overcome the deficiencies in the prior art part, in order to solve what current corn soaking method was present
The corn soaking time is short, lactic acid content is low causes the ropy phenomenon of Semen Maydis pulp, proposes a kind of thermophilic type lactobacilluss and artificial
Add the corn soaking method of the thermophilic type lactobacilluss, the method is simple to operate, easy to control, can effectively solving corn soaking
During Semen Maydis pulp it is of poor quality, effectively improve glutamic acid rate of producing acid, reduce the production method of production cost.
The purpose of the present invention is achieved through the following technical solutions：
A kind of thermophilic type lactobacilluss, entitled C15, specific name be bacillus sp., deposit number
For：CGMCC No.13471, preservation date：On December 19th, 2016, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, preservation list
Position：China Committee for Culture Collection of Microorganisms's common micro-organisms center.
And, the thermophilic type lactobacilluss are isolated from soak water of maize, and separating step is as follows：
A. plate isolation base, slant medium are prepared：Glucose 0.2%, yeast extract 1%, peptone 0.5%, bean
Dregs of rice hydrolyzed solution 0.5%, Calcium Carbonate 0.5%, agar 2%, balance of water, 121 DEG C sterilize 30 minutes, and above-mentioned percent is quality
Percent；
B. take in the corn soaking water sample access 250mL triangular flasks that 2mL conventional methods are produced, fully vibrate 15～20 points
Clock, then takes bacterium solution 1mL, moves in the test tube of the normal saline for filling 9ml mass percents 0.9%, shakes up, then from the examination
Suction 1mL is moved into and shaken up in next test tube in pipe, and with same method the 3rd test tube is diluted to, and takes 10-1、10-2、10-3Three dilute
0.1～the 0.5mL of bacterium solution of degree of releasing, is respectively applied in 2 plate isolation bases, applies uniform with Glass rod, is subsequently placed in constant temperature training
Cultivate 24～48 hours for 48～50 DEG C in foster case；
Aseptically the training of culture presevation inclined-plane is transplanted to from cultured flat board with inoculating loop extraction single bacterium colony
In foster base, after 50 DEG C are cultivated 22～24 hours, thermophilic type lactobacilluss are obtained final product, place preservation in 2～4 DEG C of refrigerators.
A kind of corn soaking method for manually adding thermophilic type lactobacilluss, with the addition of such as claim 1 in methods described
Or the thermophilic type lactobacilluss described in 2.
And, step is as follows：
(1) the mass ratio that Semen Maydiss, Semen Maydiss and soak are added in soak is 1：2.5-3.0, obtains corn soaking mixing
Liquid, and be added in corn soaking tank；
(2) soaking corn, the SO in corn soaking to 20～26h, soak2When value is between 100ppm and 200ppm, will
OD values are the seed culture fluid of 0.5～0.9 thermophilic type lactobacilluss according to seed culture fluid：The volume of corn soaking mixed liquor
Than for 0.05～0.1：1 ratio is linked in corn soaking tank, is terminated after continuing to be dipped to 45 hours, obtains corn soaking
Liquid, temperature control is at 50 DEG C ± 2 in immersion process；PH is 4.0 ± 1.0.
And, the activation method of the step (2) middle seed culture fluid is as follows：
(1) thermophilic type lactic acid bacillus mycopremna is connect from inclined-plane in seed culture medium, 50 DEG C of 18～24h of shaken cultivation, OD values
For 0.3～0.8, bacterial strain activating solution is obtained；
(2) it is by volume 0.05～0.2 by bacterial strain activating solution：1 ratio is accessed in seed culture medium, 50 DEG C of vibration trainings
18～24h is supported, seed culture fluid is obtained；
Wherein, the seed culture medium of the thermophilic type lactobacilluss is：20～35g/L of glucose, 5～20g/L of yeast extract,
5～10g/L of peptone, 5～10g/L of Semen Maydis pulp, 2～5g/L of molasses, K2HPO41.0～1.5g/L, MnSO40.00025～
0.0015g/L, VB10.0001～0.0005g/L, Na2HPO41.5～2.5g/L, solvent is water, the μ g/L of Pure biotin 0～30,
5～10g/L of Calcium Carbonate, pH are 6.5,121 DEG C and sterilize 20 minutes.
The advantage for obtaining of the invention and good effect are：
1st, method of the inventive method by adding a certain amount of thermophilic lactic acid bacteria in corn steep liquor, strengthens Corn Seeds
Grain immersion effect, shortens the corn soaking time, improves the quality of corn steep liquor, it is ensured that the high temperature sensitive type paddy of stability
The acid yield and conversion ratio of propylhomoserin fermentation, for glutamate production good working condition is provided, and reduces glutamate production cost.
2nd, the inventive method make use of thermophilic type lactobacilluss to utilize the approach product by fermentation of the saccharide in corn steep liquor
Lactogenesis acid, the lactic acid that appropriate approach by fermentation is produced is acted on and formed on corn embryosperm cell wall hole or hole, makes soaking water
Protein net is acted on into seed inside, promotes the softening and expansion of zein, moreover it is possible to make molecular mass relatively high
Soluble protein is hydrolyzed, so as to reduce soaking water evaporating concentration process in foam generation and the process characteristic of colloidal precipitation.
Method is simple to operate, easy to control.
3rd, the inventive method is the seed high density amplification culture by carrying out thermophilic type lactic acid bacteria, with reference to corn soaking mistake
Soak SO in journey2Value is between 100ppm and 200ppm, therefore addition time and addition are easily controlled, simple to operate, non-
It is very suitable for industrialized production application.
4th, the production of corn starch technique of the inventive method adopts the advanced closed circuit production procedure of wet grinding, in corn starch life
In producing the course of processing, corn soaking is slubbing operation, and sulfurous acid concentration is 0.10%～0.35% during corn soaking, soaking temperature
48～52 DEG C, 40～50h of soak time.Immersion purpose：One is that seed softens, seed aqueous 45% or so；Two is to make solubility
Material is leached, mainly mineral, protein and sugar etc.；Three is that the network structure for destroying protein makes starch and protein point
From；Four is to prevent living contaminantses, prevents spoilage organisms growth；Five be with discoloration, can inhibited oxidation enzyme reaction, it is to avoid form sediment
Powder changes colour.In immersion process, with the presence of 1.0%～1.2% a small amount of thermophilic lactic acid bacteria, using the saccharide in soak through sending out
Ferment approach produces lactic acid, and lactic acid affects to soften the Ca in seed, with soak2+、Mg2+Complex is generated, in dissolved state.It is beautiful
The soaking water obtained after rice immersion, containing dry 7%～9%, pH3.9～4.1, acidity more than 11%, microscopy lactobacilluss bacterium
Body is sturdy, and after 3～4 effect vaporizer concentrations Semen Maydis pulp is made.Semen Maydis pulp contains abundant nutrient substance, typically containing dry
>=40%, protein >=40%, acidity 9%～14%, sulfurous acid≤0.3%, concentration 22～240Be, containing biotin
(0.33mg/kg) the abundant aminoacid such as.
Specific embodiment
With reference to embodiment, the present invention is described in further detail, but embodiments of the present invention not limited to this,
Protection scope of the present invention can not be limited with following embodiments.
Experimental technique used in following embodiments if no special instructions, is conventional method；The material that used,
Reagent etc., if no special instructions, commercially obtains.
In Semen Maydis pulp used in the present invention mass fraction of corn syrup solids content be 30-50%, biological quality
Amount content is 600～800 μ g/L；The mass fraction of molasses solid content is 30-60% in molasses, and biotin mass content is
2000～3000 μ g/L.
A kind of thermophilic type lactobacilluss, entitled C15, specific name be bacillus sp., deposit number
For：CGMCC No.13471, preservation date：On December 19th, 2016, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, preservation list
Position：China Committee for Culture Collection of Microorganisms's common micro-organisms center.
Specifically, above-mentioned thermophilic type lactobacilluss are isolated from soak water of maize, and separating step is as follows：
A. plate isolation base, slant medium are prepared：Glucose 0.2%, yeast extract 1%, peptone 0.5%, bean
Dregs of rice hydrolyzed solution 0.5%, Calcium Carbonate 0.5%, agar 2%, balance of water, 121 DEG C sterilize 30 minutes, and above-mentioned percent is quality
Percent；
B. take in the corn soaking water sample access 250mL triangular flasks that 2mL conventional methods are produced, fully vibrate 15～20 points
Clock, then takes bacterium solution 1mL, moves in the test tube of the normal saline for filling 9ml mass percents 0.9%, shakes up, then from the examination
Suction 1mL is moved into and shaken up in next test tube in pipe, and with same method the 3rd test tube is diluted to, and takes 10-1、10-2、10-3Three dilute
0.1～the 0.5mL of bacterium solution of degree of releasing, is respectively applied in 2 plate isolation bases, applies uniform with Glass rod, is subsequently placed in constant temperature training
Cultivate 24～48 hours for 48～50 DEG C in foster case；
Aseptically the training of culture presevation inclined-plane is transplanted to from cultured flat board with inoculating loop extraction single bacterium colony
In foster base, after 50 DEG C are cultivated 22～24 hours, thermophilic type lactobacilluss are obtained final product, place preservation in 2～4 DEG C of refrigerators.
A kind of corn soaking method for manually adding thermophilic type lactobacilluss, with the addition of as above thermophilic in methods described
Heat type lactobacilluss.
Specifically, step is as follows：
(1) the mass ratio that Semen Maydiss, Semen Maydiss and soak are added in soak is 1：2.5-3.0, obtains corn soaking mixing
Liquid, and be added in corn soaking tank；
(2) soaking corn, the SO in corn soaking to 20～26h, soak2When value is between 100ppm and 200ppm, will
OD values are the seed culture fluid of 0.5～0.9 thermophilic type lactobacilluss according to seed culture fluid：The volume of corn soaking mixed liquor
Than for 0.05～0.1：1 ratio is linked in corn soaking tank, is terminated after continuing to be dipped to 45 hours, obtains corn soaking
Liquid, temperature control is at 50 DEG C ± 2 in immersion process；PH is 4.0 ± 1.0.
Embodiment 1：
The culture medium that example is used in this enforcement has two kinds：
(1) thermophilic type lactic acid bacteria seed culture medium：Glucose 20g/L, yeast extract 5g/L, peptone 5g/L, Semen Maydis pulp 5g/
L, molasses 2g/L, K2HPO41.0g/L, MnSO40.00025g/L, VB10.0001g/L, Na2HPO41.5g/L, solvent is water, pure
The μ g/L of biotin 5, Calcium Carbonate 5g/L.PH is 6.5,121 DEG C and sterilizes 20 minutes；
(2) temperature sensitive type glutamic acid fermentation culture medium：Amylum hydrolysate of the sugar 40g/L；Semen Maydis pulp 50ml/L；Protein hydrolyzate 10g/
L；Molasses 5ml/L；Disodium hydrogen phosphate 7g/L；Potassium chloride 2g/L；Magnesium sulfate 1.4g/L；Defoamer 0.1ml/L, solvent is water, pH
For 6.9～7.1,121 DEG C sterilize 20 minutes.
Add sugar：The Glucose Liquid that the mass concentration that sugar liquid is glucose is 600g/L is added, 115 DEG C sterilize 20 minutes.
A kind of corn soaking method for manually adding thermophilic type lactobacilluss, step is as follows：
Thermophilic type lactic acid bacteria bacillus on (1) one ring slant medium access oneself sterilizing equipped with 250ml seed culture mediums
1L triangular flask in, 50 DEG C of shaken cultivation 24h obtain bacterial strain activating solution (OD values be 0.8), are placed in cold preservation in 4 DEG C of refrigerators standby；
The bacterial strain activating solution of the thermophilic type lactic acid bacteria bacillus after by activation presses activating solution：The volume ratio of seed culture medium is 0.05～0.2：
1 ratio is accessed in seed culture medium, 50 DEG C of shaken cultivation 24h, obtains seed culture fluid (OD values are 0.8)；
(2) mass ratio that Semen Maydiss, Semen Maydiss and soak are added in soak is 1：2.5-3.0, obtains corn soaking and mixes
Liquid is closed, and is added in corn soaking tank；
(3) soaking corn, the SO in corn soaking to 20～26h, soak2When value is between 100ppm and 200ppm,
By OD values for 0.5～0.9 thermophilic type lactobacilluss seed culture fluid according to seed culture fluid：The body of corn soaking mixed liquor
Product is than being 0.05～0.1：1 ratio is linked in corn soaking tank, is terminated after continuing to be dipped to 45 hours, obtains corn soaking
Liquid, temperature control is at 50 DEG C ± 2 in immersion process；PH is 4.0 ± 1.0；
(4) by corn steep liquor Jing after evaporation and concentration, temperature sensitive type bacterial strain glutamic acid fermentation Semen Maydis pulp is produced into.
The Semen Maydis pulp that said method is produced is applied to into the method that temperature sensitive type glutamic acid fermentation culture medium produces glutamic acid,
Step is as follows：
To produce bacterium seed liquor (OD is 0.8～1.2) by 15% inoculum concentration and access fermentation medium, using continuous stream plus
The mode of the ammonia of mass fraction 28% is supplemented nitrogen source and controls the pH of fermentation liquid 6.8～7.2, is flowed as needed plus appropriate bubble
Oppose to lather collapse, stream adds the glucose solution (adding sugar liquid) that mass concentration is 600g/L residual sugar to be controlled in fermentation liquid
Mass fraction be 5～10g/L, fermentation culture to after fermentation in 32 hours terminates, that is, ferment to obtain glutamic acid；
Wherein, the microbial strains for being used are temperature sensitive type glutamic acid producer；
Fermentation medium：Amylum hydrolysate of the sugar 40g/L；Semen Maydis pulp 50ml/L；Protein hydrolyzate 10g/L；Molasses 5ml/L；Phosphorus
Sour disodium hydrogen 7g/L；Potassium chloride 2g/L；Magnesium sulfate 1.4g/L；Defoamer 0.1ml/L, solvent is water, and pH is 6.9～7.1,121
DEG C sterilizing 20 minutes；
Add sugar liquid：The Glucose Liquid that the mass concentration that sugar liquid is glucose is 600g/L is added, 115 DEG C sterilize 20 minutes
It is standby afterwards.
After testing, it is 180g/L that glutamate levels are produced in fermentation, and saccharic acid conversion ratio is 68%.
Embodiment 2：
The culture medium that example is used in this enforcement has two kinds：
(1) thermophilic lactic acid bacteria seed culture medium：Glucose 20g/L, yeast extract 10g/L, peptone 5g/L, Semen Maydis pulp 5g/L,
Molasses 2g/L, K2HPO41.0g/L, MnSO40.00025g/L, VB10.0001g/L, Na2HPO41.5g/L, solvent is water, pure life
Thing 5 μ g/L of element, Calcium Carbonate 5g/L.PH is 6.5,115 DEG C and sterilizes 20 minutes；
(2) temperature sensitive type glutamic acid fermentation culture medium：Amylum hydrolysate of the sugar 50g/L；Semen Maydis pulp 55ml/L；Protein hydrolyzate 15g/
L；Molasses 10ml/L；Disodium hydrogen phosphate 8g/L；Potassium chloride 3g/L；Magnesium sulfate 1.5g/L；Defoamer 0.2ml/L, solvent is water, pH
For 6.9～7.1,121 DEG C sterilize 20 minutes.
Add sugar：The Glucose Liquid that the mass concentration that sugar liquid is glucose is 600g/L is added, 115 DEG C sterilize 20 minutes.
A kind of corn soaking method for manually adding thermophilic type lactobacilluss, with the addition of as above thermophilic in methods described
Heat type lactobacilluss.
Specifically, step is as follows：
(1) the mass ratio that Semen Maydiss, Semen Maydiss and soak are added in soak is 1：2.5-3.0, obtains corn soaking mixing
Liquid, and be added in corn soaking tank；
(2) soaking corn, the SO in corn soaking to 20～26h, soak2When value is between 100ppm and 200ppm, will
OD values are the seed culture fluid of 0.5～0.9 thermophilic type lactobacilluss according to seed culture fluid：The volume of corn soaking mixed liquor
Than for 0.05～0.1：1 ratio is linked in corn soaking tank, is terminated after continuing to be dipped to 45 hours, obtains corn soaking
Liquid, temperature control is at 50 DEG C ± 2 in immersion process；PH is 4.0 ± 1.0；
The activation method of the step (2) middle seed culture fluid is as follows：
(1) thermophilic type lactic acid bacillus mycopremna is connect from inclined-plane in seed culture medium, 50 DEG C of 18～24h of shaken cultivation, OD values
For 0.3～0.8, bacterial strain activating solution is obtained；
(2) it is by volume 0.05～0.2 by bacterial strain activating solution：1 ratio is accessed in seed culture medium, 50 DEG C of vibration trainings
18～24h is supported, seed culture fluid is obtained；
Wherein, the seed culture medium of the thermophilic type lactobacilluss is：20～35g/L of glucose, 5～20g/L of yeast extract,
5～10g/L of peptone, 5～10g/L of Semen Maydis pulp, 2～5g/L of molasses, K2HPO41.0～1.5g/L, MnSO40.00025～
0.0015g/L, VB10.0001～0.0005g/L, Na2HPO41.5～2.5g/L, solvent is water, the μ g/L of Pure biotin 0～30,
5～10g/L of Calcium Carbonate, pH are 6.5,121 DEG C and sterilize 20 minutes.
(3) by corn steep liquor Jing after evaporation and concentration, temperature sensitive type bacterial strain glutamic acid fermentation Semen Maydis pulp is produced into.
The Semen Maydis pulp produced using said method is applied to the side that temperature sensitive type glutamic acid fermentation culture medium produces glutamic acid
Method, step is as follows：
Seed liquor (OD is 0.8～1.2) is accessed into fermentation medium by 15% inoculum concentration production bacterium, using continuous stream plus
The mode of the ammonia of mass fraction 28% is supplemented nitrogen source and controls the pH of fermentation liquid 6.8～7.2；Stream plus in right amount bubble as needed
Oppose to lather collapse, stream adds the glucose solution (adding sugar liquid) that mass concentration is 700g/L residual sugar to be controlled in fermentation liquid
Mass fraction be 5～10g/L, fermentation culture to after fermentation in 32 hours terminates, that is, ferment to obtain glutamic acid；
Wherein, the microbial strains for being used are temperature sensitive type glutamic acid producer；
Fermentation medium：Amylum hydrolysate of the sugar 50g/L；Semen Maydis pulp 55ml/L；Protein hydrolyzate 15g/L；Molasses 10ml/L；Phosphorus
Sour disodium hydrogen 8g/L；Potassium chloride 3g/L；Magnesium sulfate 1.5g/L；Defoamer 0.2ml/L, solvent is water, and pH is 6.9～7.1,121
DEG C sterilizing 20 minutes；
Add sugar liquid：The Glucose Liquid that the mass concentration that sugar liquid is glucose is 700g/L is added, 115 DEG C sterilize 20 minutes
It is standby afterwards.
After testing, it is 191g/L that glutamate levels are produced in fermentation, and saccharic acid conversion ratio is 70%.
Embodiment 3：
Culture medium used in the present embodiment has two kinds：
(1) thermophilic lactic acid bacteria seed culture medium：Glucose 30g/L, yeast extract 10g/L, peptone 5g/L, Semen Maydis pulp 5g/
L, molasses 2g/L, K2HPO41.0g/L, MnSO40.00025g/L, VB10.0001g/L, Na2HPO41.5g/L, solvent is water, pure
The μ g/L of biotin 5, Calcium Carbonate 5g/L.PH is 6.5,115 DEG C and sterilizes 20 minutes；
(2) temperature sensitive type glutamic acid fermentation culture medium：Amylum hydrolysate of the sugar 60g/L；Semen Maydis pulp 60ml/L；Protein hydrolyzate 20g/
L；Molasses 15ml/L；Disodium hydrogen phosphate 9g/L；Potassium chloride 4g/L；Magnesium sulfate 1.6g/L；Defoamer 0.3ml/L, solvent is water, pH
For 6.9～7.1,121 DEG C sterilize 20 minutes.
Add sugar：The Glucose Liquid that the mass concentration that sugar liquid is glucose is 600g/L is added, 115 DEG C sterilize 20 minutes.
A kind of corn soaking method for manually adding thermophilic type lactobacilluss and Semen Maydis pulp are produced, and step is with embodiment 2.
The Semen Maydis pulp produced using said method is applied to the side that temperature sensitive type glutamic acid fermentation culture medium produces glutamic acid
Method, step is as follows：
To produce bacterium seed liquor (OD is 0.8～1.2) by 15% inoculum concentration and access fermentation medium, using continuous stream plus
The mode of the ammonia of mass fraction 28% is supplemented nitrogen source and controls the pH of fermentation liquid 6.8～7.2；Stream plus in right amount bubble as needed
Oppose to lather collapse, stream adds the glucose solution (adding sugar liquid) that mass concentration is 800g/L residual sugar to be controlled in fermentation liquid
Mass fraction be 5～10g/L, fermentation culture to after fermentation in 32 hours terminates, that is, ferment to obtain glutamic acid；
Wherein, the microbial strains for being used are temperature sensitive type glutamic acid producer；
Fermentation medium：Amylum hydrolysate of the sugar 60g/L；Semen Maydis pulp 60ml/L；Protein hydrolyzate 20g/L；Molasses 15ml/L；Phosphorus
Sour disodium hydrogen 9g/L；Potassium chloride 4g/L；Magnesium sulfate 1.6g/L；Defoamer 0.3ml/L, solvent is water, and pH is 6.9～7.1,121
DEG C sterilizing 20 minutes；
Add sugar liquid：The Glucose Liquid that the mass concentration that sugar liquid is glucose is 800g/L is added, 115 DEG C sterilize 20 minutes
It is standby afterwards.
After testing, it is 208g/L that glutamate levels are produced in fermentation, and saccharic acid conversion ratio is 72%.
Using culture medium fermenting and producing glutamic acid of the present invention and using known culture medium fermenting and producing glutamic acid
Technical specification contrast is shown in Table 1:
The technology of the culture medium fermenting and producing glutamic acid of the present invention of table 1 and the glutamic acid using known culture medium fermenting and producing
Index contrast table
Can be seen that by the result of table 1 carries out the side of the Semen Maydis pulp that corn soaking is produced with artificial addition thermophilic lactic acid bacteria
Method, every production target of temperature sensitive type glutamic acid fermentation does not have significantly reduction, and glutamic acid acid production rate and saccharic acid conversion ratio also have
Improved.It is feasible to prove that the inventive method is popularized in an all-round way in temperature sensitive type glutamic acid fermentation technique productions.
Claims (5)
1. a kind of thermophilic type lactobacilluss, it is characterised in that：Entitled C15, specific name is bacillus sp.,
Deposit number is：CGMCC No.13471, preservation date：On December 19th, 2016, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Number, depositary institution：China Committee for Culture Collection of Microorganisms's common micro-organisms center.
2. thermophilic type lactobacilluss according to claim 1, it is characterised in that：The thermophilic type lactobacilluss are isolated from jade
In rice soaking water, separating step is as follows：
A. plate isolation base, slant medium are prepared：Glucose 0.2%, yeast extract 1%, peptone 0.5%, bean cake water
Solution liquid 0.5%, Calcium Carbonate 0.5%, agar 2%, balance of water, 121 DEG C sterilize 30 minutes, and above-mentioned percent is quality percentage
Number；
B. take in the corn soaking water sample access 250mL triangular flasks that 2mL conventional methods are produced, fully vibrate 15～20 minutes,
Then bacterium solution 1mL is taken, is moved in the test tube of the normal saline for filling 9ml mass percents 0.9%, shaken up, then from the test tube
Middle suction 1mL is moved into and shaken up in next test tube, and with same method the 3rd test tube is diluted to, and takes 10-1、10-2、10-3Three dilutions
0.1～the 0.5mL of bacterium solution of degree, is respectively applied in 2 plate isolation bases, applies uniform with Glass rod, is subsequently placed in constant temperature culture
Cultivate 24～48 hours for 48～50 DEG C in case；
Aseptically culture presevation slant medium is transplanted to from cultured flat board with inoculating loop extraction single bacterium colony
It is interior, after 50 DEG C are cultivated 22～24 hours, thermophilic type lactobacilluss are obtained final product, place preservation in 2～4 DEG C of refrigerators.
3. a kind of corn soaking method for manually adding thermophilic type lactobacilluss, it is characterised in that：With the addition of in methods described as
Thermophilic type lactobacilluss described in claim 1 or 2.
4. the corn soaking method for manually adding thermophilic type lactobacilluss according to claim 3, it is characterised in that：Step
It is as follows：
(1) the mass ratio that Semen Maydiss, Semen Maydiss and soak are added in soak is 1：2.5-3.0, corn soaking mixed liquor is obtained,
And be added in corn soaking tank；
(2) soaking corn, the SO in corn soaking to 20～26h, soak2When value is between 100ppm and 200ppm, by OD values
For 0.5～0.9 thermophilic type lactobacilluss seed culture fluid according to seed culture fluid：The volume ratio of corn soaking mixed liquor is
0.05～0.1：1 ratio is linked in corn soaking tank, is terminated after continuing to be dipped to 45 hours, obtains corn steep liquor, is soaked
Temperature control is at 50 DEG C ± 2 during bubble；PH is 4.0 ± 1.0.
5. the corn soaking method for manually adding thermophilic type lactobacilluss according to claim 4, it is characterised in that：It is described
The activation method of step (2) middle seed culture fluid is as follows：
(1) thermophilic type lactic acid bacillus mycopremna is connect from inclined-plane in seed culture medium, 50 DEG C of 18～24h of shaken cultivation, OD values are
0.3～0.8, obtain bacterial strain activating solution；
(2) it is by volume 0.05～0.2 by bacterial strain activating solution：1 ratio is accessed in seed culture medium, 50 DEG C of shaken cultivation 18
～24h, obtains seed culture fluid；
Wherein, the seed culture medium of the thermophilic type lactobacilluss is：20～35g/L of glucose, 5～20g/L of yeast extract, albumen
5～10g/L of peptone, 5～10g/L of Semen Maydis pulp, 2～5g/L of molasses, K2HPO41.0～1.5g/L, MnSO40.00025～
0.0015g/L, VB10.0001～0.0005g/L, Na2HPO41.5～2.5g/L, solvent is water, the μ g/ of Pure biotin 0～30
L, 5～10g/L of Calcium Carbonate, pH are 6.5,121 DEG C and sterilize 20 minutes.
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| CN115669854A (en) * | 2022-11-16 | 2023-02-03 | 吉林农业大学 | High-processing-quality high-nutrition corn flour and efficient industrial milling method and application thereof |
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| CN109234185A (en) * | 2018-06-24 | 2019-01-18 | 山东省科学院生物研究所 | A kind of agent of lactic acid bacteria composition being directly used in wet process corn soaking |
| CN109234185B (en) * | 2018-06-24 | 2020-03-31 | 山东省科学院生物研究所 | Lactic acid bacteria agent composition directly used for wet corn soaking |
| CN109913388A (en) * | 2019-03-20 | 2019-06-21 | 中粮集团有限公司 | Improve composite bacteria agent and its application of corn soaking effect |
| CN110904163A (en) * | 2019-12-31 | 2020-03-24 | 内蒙古阜丰生物科技有限公司 | Method for improving lactic acid content of corn steep liquor |
| CN114181980A (en) * | 2021-11-09 | 2022-03-15 | 吉林中粮生化有限公司 | Corn steep liquor, preparation method and application thereof, and production method of high-optical-purity lactic acid |
| CN115669854A (en) * | 2022-11-16 | 2023-02-03 | 吉林农业大学 | High-processing-quality high-nutrition corn flour and efficient industrial milling method and application thereof |
| CN115669854B (en) * | 2022-11-16 | 2024-05-24 | 吉林农业大学 | Corn flour and industrialized flour making method and application thereof |
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