CN102649971A - Method for cultivating aspergillus niger mouldy bran and method for preparing citric acid through fermentation - Google Patents
Method for cultivating aspergillus niger mouldy bran and method for preparing citric acid through fermentation Download PDFInfo
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- CN102649971A CN102649971A CN2012101501126A CN201210150112A CN102649971A CN 102649971 A CN102649971 A CN 102649971A CN 2012101501126 A CN2012101501126 A CN 2012101501126A CN 201210150112 A CN201210150112 A CN 201210150112A CN 102649971 A CN102649971 A CN 102649971A
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Abstract
The invention discloses a method for cultivating aspergillus niger mouldy bran. The method for cultivating the aspergillus niger mouldy bran comprises the following steps: sterilizing a culture medium; inoculating aspergillus niger into the sterilized culture medium; and cultivating the mouldy bran, wherein the culture medium comprises corn fibrous residues. The invention further discloses a method for preparing citric acid through fermentation. The method for preparing the citric acid through fermentation comprises the following steps: inoculating the aspergillus niger mouldy bran obtained by the method for cultivating the aspergillus niger mouldy bran into a citric acid seed culture medium; performing seed culture; inoculating the seed culture liquid into a citric acid fermentation culture medium; and performing fermentation culture. According to the method for cultivating the aspergillus niger mouldy bran, the culture cost of the aspergillus niger mouldy bran can be reduced and the quality and the stability of the aspergillus niger mouldy bran can be improved, so that the quality and the stability of the subsequent citric acid fermentation are improved.
Description
Technical field
The present invention relates to a kind of cultural method of black mold wheat bran, and the black mold wheat bran that uses this cultural method to obtain carries out the fermentative prepn methods of citric acid.
Background technology
Hydrocerol A is first acid in the organic acid, because the excellent properties of aspects such as physics, chemistry is widely used in industrial circles such as medicine, chemistry, electronics, weaving, oil, leather, building, photography, plastics, casting and pottery.
At present the preparation method of Hydrocerol A black mold seed generally comprises: Testa Tritici is carried out pre-treatment, carries out sterilising treatment after adorning three grades of flasks, and insert inclined-plane black mold seed and cultivate and obtain qualified wheat bran, be further used for fermentative prodn.But it is bigger to the influence of wheat bran seed quality that the back is cultivated in the inoculation of use Testa Tritici, and, also may cause the performance difference of wheat bran seed bigger, and then the instability that causes follow-up citric acid fermentation to be produced.
Summary of the invention
The objective of the invention is to overcome the lower and quality problem of unstable of wheat bran quality of cultivating, provide a mass higher, the stable cultural method of black mold wheat bran preferably through above-mentioned Testa Tritici.
The black mold wheat bran that the present invention also aims to provide a kind of cultural method that uses above-mentioned black mold wheat bran to obtain carries out the fermentative prepn methods of citric acid.
Contriver of the present invention finds, adopts the performance difference of the wheat bran that obtains through pretreated Testa Tritici cultivation bigger, and when being applied to citric acid fermentation, the quality of Hydrocerol A is also unstable.Infer reason maybe for: in the method that adopts Testa Tritici inoculation culture wheat bran; Said pretreated method is generally water Testa Tritici is repeatedly eluriated; Under the prerequisite of taking all factors into consideration cost and effect, generally believe that keeping its starch content is that 3-8 weight % promptly can satisfy culture requirement.But, eluriate amount of starch stable that step is difficult to control the Testa Tritici of each batch, not only wasted water resources thus, also increased cost, but also bigger, and then caused the instability of follow-up fermentative prodn the influence of wheat bran seed quality.Supposition to the bigger reason of the influence of wheat bran seed quality maybe for: 1) eluriate the loss that step causes the soluble protein (being nitrogenous source) that aspergillus niger strain utilizes easily; And residual protein aspergillus niger strain is not easy to utilize, thereby has increased the difficulty that aspergillus niger strain utilizes nitrogenous source; 2) starch content is unfixing in the elutriation process, causes wheat bran to meet group and causes nitrogenous source not make full use of.In addition, contriver of the present invention is unexpected to be found, adopts the substratum that contains the zein fiber slag that aspergillus niger strain is carried out wheat bran and cultivates, and can access the high and good black mold wheat bran of quality stability of a kind of quality, thereby accomplish the present invention.
In the present invention, adopt the substratum that contains the zein fiber slag that aspergillus niger strain is carried out wheat bran and cultivate, can access the high and good black mold wheat bran of quality stability of a kind of quality.Infer that reason possibly be the 1) stable components in the zein fiber slag, and need not through eluriating step that soluble protein does not run off in the zein fiber slag, aspergillus niger strain utilizes the protein in the zein fiber slag easily; 2) with respect to the fiber of Testa Tritici, the fiber in the zein fiber slag is utilized by aspergillus niger strain more easily.
That is, the invention provides a kind of cultural method of black mold wheat bran, this method comprises sterilizes substratum, in culture medium after sterilization, inserts aspergillus niger strain and carries out the wheat bran cultivation, and wherein, said substratum contains the zein fiber slag.
The present invention also provides a kind of fermentative prepn methods of citric acid; Wherein, This method comprise with the resulting black mold wheat bran of cultural method that adopts above-mentioned black mold wheat bran be linked into carry out seed culture in the Hydrocerol A seed culture medium after, the seed culture fluid that obtains is linked into carries out fermentation culture in the citric acid fermentation substratum again.
According to the cultural method of black mold wheat bran of the present invention, this method can reduce the cultivation cost of black mold wheat bran, and can improve the quality and the stability of black mold wheat bran, and then improves the quality and the stability of follow-up citric acid fermentation.
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.
According to the cultural method of black mold wheat bran of the present invention, this method comprises sterilizes substratum, in culture medium after sterilization, inserts aspergillus niger strain and carries out the wheat bran cultivation, and wherein, said substratum contains the zein fiber slag.
According to the present invention, under the preferable case, said zein fiber slag is the by product zein fiber slag that obtains in the W-Gum production process.In the present invention, owing to can utilize the sub product zein fiber slag in the W-Gum production process, and this zein fiber slag need not to eluriate, and therefore can reduce cost.
According to the present invention; Because relating generally to, the cultural method of black mold wheat bran provided by the invention adopt the preparation of zein fiber slag to be used for the substratum that wheat bran is cultivated; Therefore, do not limit for the preparation of zein fiber slag is special, as long as preferably satisfy each component concentration requirement in the zein fiber slag.For example, can be the zein fiber slag by product that obtains in the W-Gum production process.The production of W-Gum (being W-Gum technology) can be carried out with reference to prior art.This W-Gum technology generally includes corn soaking, takes off plumule, grind and obtain coarse starch milk and zein fiber slag crust.
Particularly, in the present invention, the zein fiber slag can obtain through following method:
At first, corn particle is soaked.The condition that corn particle is soaked can in very large range change; But under the preferable case; The temperature of soaking can be 40-60 ℃, has no particular limits for the ratio between the amount of soak solution and dried corn particle, as long as dried corn particle is fully soaked; But under the preferable case, the liquid level of said soak solution is higher than at least 10 centimetres of dried corn particles.Said soak solution can be the sulfurous acid solution of 0.15-0.25 weight % concentration, and soak time can be preferably 55-65 hour for more than 50 hours.According to the present invention, it is conventionally known to one of skill in the art soaking the equipment that is adopted, and for example, can in steeping tank, soak corn.
Secondly, remove the plumule of the corn particle after the immersion.Remove the method that the method for the plumule of the corn particle after the immersion can be taken off embryo for various corns; For example; This method can comprise: will once grind through the corn particle after soaking; The condition of once grinding makes corn particle be broken into average particle diameter can be the particle of 2-5 millimeter, and removes the plumule under grinding; Carry out regrind afterwards, the condition of regrind makes the particulate average particle diameter that obtains can be the 1-1.5 millimeter, and removes the plumule under grinding.
Then, isolate the zein fiber slag.The method of isolating the zein fiber slag is conventionally known to one of skill in the art, for example, can obtain soup compound with carrying out fine grinding except that the corn particle behind the degerming; This soup compound is incorporated into sieves the after scouring solid in the pressure curved sieve, obtain the zein fiber slag.The compass screen surface radian of above-mentioned bent sieve is 100-120 °, and the sieve seam width of bent sieve is the 50-75 micron, and feed pressure is 0.2-0.4MPa.Said pressure curved sieve can be commercially available, for example the DZQ50 type pressure curved sieve of Yixing starch instrument factory production.In addition, the soup compound of isolating the zein fiber slag obtains milk of starch after removing Deproteinization.
According to the present invention, contain zein fiber, water, starch and protein in the zein fiber slag that obtains through aforesaid method.Under the preferred situation, in the said zein fiber slag, the content of zein fiber is 20-35 weight %, and the content of water is 45-65 weight %, and starch content is 1.5-5 weight %, and protein contnt is 5-15 weight %; In the more preferably said zein fiber slag, the content of zein fiber is 25-30 weight %, and the content of water is 54-60 weight %, and starch content is 2-4 weight %, and protein contnt is 7-12 weight %.The impurity that possibly also contain in addition, other equal amounts in the said zein fiber slag.
According to the present invention, in the zein fiber slag that obtains through aforesaid method, its protein contnt insufficient nitrogen might occur during less than 5 weight %, influences training quality.In this case, can therefore, can also contain various nitrogenous source known in those skilled in the art in the said substratum through in substratum, adding the problem that the mode of replenishing nitrogenous source solves insufficient nitrogen.The addition of said additional nitrogenous source can come according to the nitrogenous source content (coming from the protein in the zein fiber slag) of said zein fiber slag suitably to select, as long as preferably can make the scope of the nitrogenous source content of substratum at 0.7-1.2 weight %.Said additional nitrogenous source can be in corn steep liquor, urea, ammonium sulfate and an ammonium nitrate etc. one or more, is preferably corn steep liquor.Said corn steep liquor is meant the corn soaking liquid that contains soluble sugar that obtains behind the soaking corn particle in the above-mentioned acquisition zein fiber slag method.In order to reduce the use volume of said corn steep liquor; Re-use after preferably this corn steep liquor being concentrated; Volume to after concentrating does not have special requirement, generally speaking, doubly gets final product through the 0.5-0.1 that this corn steep liquor is evaporated to original volume under 35-80 ℃.
In addition, can also contain in the said substratum and cultivate needed other trace elements that well known to a person skilled in the art of black mold wheat bran, for example, sodium, potassium, oxygen etc., the visual particular case of addition and selecting here repeats no more.
According to the present invention, the access amount of aspergillus niger strain can be the routine access amount of this area when cultivating the black mold wheat bran.For example, be benchmark with every gram substratum, the access amount of said aspergillus niger strain is 1.5 * 10
5~ 5.5 * 10
5Individual aspergillus niger spore; Under the preferable case, be benchmark with every gram substratum, the access amount of said aspergillus niger strain is 2 * 10
5~ 4 * 10
5Individual aspergillus niger spore.
According to the present invention, carry out the wheat bran culture condition and can be this area condition commonly used when cultivating the black mold wheat bran.For example, this culture condition comprises: temperature is 30-37 ℃, and humidity is 45-55%, and air flow is the 0.1-0.3 volume: (volume minute), and incubation time is 7-15 days; More preferably temperature is 32-35 ℃, and humidity is 47-53%, and air flow is the 0.1-0.2 volume: (volume minute), incubation time is 9-11 days
In the present invention; Term " air flow " is generally with ventilation expression recently; Usually recently to represent (V/Vmin) through the volume of air of unit volume nutrient solution in the PM, for example ventilation is than being 1:0.1-1, and being called for short air flow is the 0.01-1 volume: (volume minute).
According to the present invention, the said method that substratum is sterilized can adopt and well known to a person skilled in the art the whole bag of tricks.For example, can be that 121-125 ℃, pressure are sterilization 45-90min under the 0.1-0.15MPa in temperature with substratum.
The present invention also provides a kind of fermentative prepn methods of citric acid; Wherein, This method comprise with the resulting black mold wheat bran of cultural method that adopts above-mentioned black mold wheat bran be linked into carry out seed culture in the Hydrocerol A seed culture medium after, the seed culture fluid that obtains is linked into carries out fermentation culture in the citric acid fermentation substratum again.
According to the present invention, said Hydrocerol A seed culture medium there is not special requirement, as long as can be used for the substratum of Hydrocerol A seed culture.Under the preferable case, said Hydrocerol A seed culture medium is a starchy material enzymatic liquefaction clear liquid.Total amount with this starchy material enzymatic liquefaction clear liquid is a benchmark, and the total reducing sugar in the starchy material enzymatic liquefaction clear liquid is that 7-13 weight %, nitrogenous source (in the nitrogen element) are 0.1-0.5 weight %; Preferably, the total reducing sugar in this starchy material enzymolysis solution is that 9-11 weight %, nitrogenous source are 0.1-0.3 weight %.In addition, as required, also can in seed culture medium, add and replenish nitrogenous source.The addition of said additional nitrogenous source can be the 0.01-0.05 weight % of substratum gross weight.The kind of said additional nitrogenous source is conventionally known to one of skill in the art, and for example, said additional nitrogenous source can be in urea, ammonium sulfate and an ammonium nitrate etc. one or more.
According to the present invention, to the citric acid fermentation substratum do not have a special requirement yet, as long as can be used for the fermention medium of citric acid fermentation.Under the preferable case, said fermention medium contains the starchy material enzymolysis product.Total amount with this starchy material enzymolysis product is a benchmark, and the total reducing sugar in the starchy material enzymolysis product is that 14.5-18 weight %, nitrogenous source (in the nitrogen element) are 0.05-0.2 weight %; Preferably, the total reducing sugar in this starchy material enzymolysis solution is that 15.5-17 weight %, nitrogenous source are 0.08-0.15 weight %.Usually; The starchy material enzymolysis obtains starchy material enzymolysis residue and starchy material enzymatic liquefaction clear liquid; Usually can starchy material enzymatic liquefaction clear liquid be used to prepare Hydrocerol A seed culture medium or citric acid fermentation substratum, also can with starchy material enzymatic liquefaction clear liquid be used to prepare the citric acid fermentation substratum after starchy material enzymolysis residue mixes.Citric acid fermentation substratum according to the invention (being the starchy material enzymolysis product) is preferably mixed with water or is not mixed with water by starchy material enzymolysis residue and starchy material enzymatic liquefaction clear liquid and obtains; And further preferred gross weight with said fermention medium is that 100 weight parts are benchmark; The consumption of said starchy material enzymatic liquefaction clear liquid is the 70-90 weight part; The consumption of said starchy material enzymolysis residue is the 10-20 weight part, and the consumption of water is 0-10 amount part.
According to the present invention, said starchy material enzymatic liquefaction clear liquid can prepare through several different methods, for example; Can prepare through following method: starchy material is pulverized, the product after pulverizing is carried out enzymolysis, obtain enzymolysis product; With the enzymolysis product solid-liquid separation; Obtain starchy material enzymatic liquefaction clear liquid and starchy material enzymolysis residue, it is 5-60 weight %, more preferably 30-50 weight % that the condition of said solid-liquid separation makes the solid content of starchy material enzymolysis residue.
According to the present invention; Said starchy material can be the known various raw materials that contain starch that can be used for enzymolysis, fermentative prepn Hydrocerol A of ability, for example, can be selected from corn, potato class (like cassava) and the wheat one or more; Under the preferable case, said starchy material is a corn.
Said enzymolysis step can be accomplished through this area method commonly used, and such as in crushed products, adding microbes producing cellulase and/or enzyme, insulation is accomplished under the growth temperature of microbes producing cellulase and/or the great-hearted temperature of enzyme.Said microbes producing cellulase be can secreting amylase microbes producing cellulase.Said enzyme comprises glycase.
Because microorganism growth can produce by product, the therefore preferred enzyme that directly adds.The consumption of said enzyme is The more the better, from cost consideration, and the dry weight basis of the crushed products after preferably pulverizing with every gram, said diastatic consumption is a 15-50 enzyme activity unit.
Being defined as of the enzyme activity unit of enzyme according to the 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.
The temperature of said enzymolysis can in very large range change, and is preferably 70-105 ℃, more preferably 80-95 ℃.The longer the better on the time theory of said enzymolysis, considers plant factor, and the time of preferred said enzymolysis is 90-150 minute, more preferably 100-120 minute.The pH value of said enzymolysis can in very large range change, and is preferably 5.0-7.0, and more preferably the pH value is 5.4-5.7.
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, saccharifying enzyme and isoamylase.
According to the present invention, preferably use AMS and/or isoamylase.
According to the present invention, the method and apparatus of said solid-liquid separation is conventionally known to one of skill in the art, for example, and pressure filter or whizzer.
According to the present invention, when carrying out the Hydrocerol A seed culture, the access amount of aspergillus niger strain can be the routine access amount of this area.For example, be benchmark with every milliliter of lemon acid seed culture medium, the access amount of black mold wheat bran makes that the access amount of aspergillus niger spore is 1.5 * 10
5~ 4.5 * 10
5Individual; Preferably, be benchmark with every milliliter of Hydrocerol A seed culture medium, the access amount of black mold wheat bran makes that the access amount of aspergillus niger spore is 2 * 10
5~ 3 * 10
5Individual.
In the present invention, aspergillus niger spore is that the form with the black mold wheat bran is linked in the substratum, and the aspergillus niger spore number in the black mold wheat bran can record according to the blood cell colony counting method.Particularly; This method is: take by weighing 5 groups in 1g black mold wheat bran, add in every group and contain the SPSS of telling anthracene 4%, make spore become unbound state through magnetic stirring apparatus and UW; Carry out the spore counting at microscopically respectively, get the MV of 5 groups of data that obtain.
In the present invention; The degree of Hydrocerol A seed culture can be measured through sampling sediments microscope inspection, acid test and pH and observe the growth of black mold, when pH 2.0-2.5, acidity 0.5-2.0%, bacterium ball size evenly, mycelia is sturdy stops to cultivate when stretching out.
The condition of above-mentioned seed culture can in very large range change; For example said culture condition can comprise: the temperature of cultivation can be 33-42 ℃; The pH value can be 2-6, and air flow can be the 0.1-0.5 volume: (volume minute), and the time of cultivation can be 18-40 hour; Under the preferred situation, said culture condition can comprise: the temperature of cultivation can be 34-38 ℃, and the pH value can be 2.5-4.5, and air flow can be the 0.15-0.4 volume: (volume minute), the time of said cultivation can be 22-30 hour.
According to the present invention, when carrying out the citric acid fermentation cultivation, the access amount of Hydrocerol A seed culture fluid can be the routine access amount of this area.For example, be benchmark with every milliliter of citric acid fermentation substratum, the access amount of seed culture fluid makes that the access amount of aspergillus niger spore is 1 * 10
4~ 5 * 10
4Individual; Preferably, be benchmark with every milliliter of citric acid fermentation substratum, the access amount of seed culture fluid makes that the access amount of aspergillus niger spore is 1.5 * 10
4~ 3.5 * 10
4Individual.
According to the present invention, the condition of citric acid fermentation can be the normal condition of this area.For example the condition of citric acid fermentation can comprise: the temperature of cultivation can be 33-40 ℃, and the pH value can be 1.5-6, and air flow can be the 0.1-0.5 volume: (volume minute), and the time of cultivation can be 45-65 hour; More preferably said culture condition can comprise: the temperature of cultivation can be 34-39 ℃, and the pH value can be 1.5-3.0, and air flow can be the 0.12-0.3 volume: (volume minute), the time of said cultivation can be 50-60 hour.
The tunning Hydrocerol A for preparing according to method of the present invention can be used conventional method, separate and refining according to the requirement of different Industrial products, such as neutralization, acidolysis, decolouring, concentrate, crystallization, packing.
Below will describe the present invention through embodiment.
In following examples, the measuring method of each component content of zein fiber slag is:
Zein fiber Determination on content in the zein fiber slag: measure zein fiber content in the zein fiber slag according to GB-T 6434-1994.
The mensuration of water-content in the zein fiber slag: take by weighing 10g zein fiber slag, 90 ℃ of down oven dry, the weight after the oven dry and the ratio of the weight of the zein fiber slag that takes by weighing are the content of the water in the zein fiber slag.
The mensuration of starch content in the zein fiber slag: measure starch content in the zein fiber slag according to GB/T 5009.9-2008.
The mensuration of protein contnt in the zein fiber slag: adopt nitrogen determination to measure protein contnt in the zein fiber slag.
In the Comparative Examples, the measuring method of each method for measuring components of Testa Tritici is:
The mensuration of fibre content in the Testa Tritici: measure fibre content in the Testa Tritici according to GB-T 6434-1994.
The mensuration of water-content in the Testa Tritici: take by weighing the 10g Testa Tritici, 90 ℃ of oven dry down, the per-cent of the weight of weight after the oven dry and the Testa Tritici that takes by weighing is the content of the water in the Testa Tritici.
The mensuration of starch content in the Testa Tritici: measure starch content in the Testa Tritici according to GB/T 5009.9-2008.
The mensuration of protein contnt in the Testa Tritici: adopt nitrogen determination to measure protein contnt in the Testa Tritici.
In following examples, the aspergillus niger spore number in the black mold wheat bran can record according to the blood cell colony counting method.Particularly; This method is: take by weighing 5 groups in 1g black mold wheat bran, add in every group and contain the SPSS of telling anthracene 4%, make spore become unbound state through magnetic stirring apparatus and UW; Carry out the spore counting at microscopically respectively, get the MV of 5 groups of data that obtain.
Corn enzymolysis clear liquid in following examples and corn enzymolysis residue can obtain with reference to following method:
(1) 56 kg corn that will gather in the crops are stewing in hot water tank profit, are 15 weight % until the water cut of corn, pulverize then, obtain average particle diameter and be 400 microns pulverizing after product.
(2) product after will pulverizing is sized mixing by the concentration of 25 weight %; With respect to the product after every gram pulverizing, add glycase (Novozymes Company, the AMS of 20 enzyme activity units; Equal glycase for this reason in the embodiment of the invention); Getting into injector, is enzymolysis 100 minutes under 5.5 the condition at 85 ℃, pH, obtains enzymolysis product.
(3) with enzymolysis product through carrying out press filtration with the fluid pressure type plate-and-frame filter press, isolate enzymatic liquefaction clear liquid and enzymolysis filter residue, wherein, the water cut of enzymolysis residue is 50%.
Concentration (abbreviation acidity) according to gained citric acid fermentation broth in GB 1987-2007 standard detection following examples; Calculate the transformation efficiency of Hydrocerol A, weight * 100% of the volume/total reducing sugar of the concentration of transformation efficiency (%)=citric acid fermentation broth (abbreviation acidity) * citric acid fermentation broth.
In following examples, fermentation grain consumption is meant the amount of producing 1 ton of corn that Hydrocerol A consumes.Ferment strength per hour is meant in the production that every cubic metre of fermented liquid produces the amount of acid.
Embodiment 1
1) preparation of zein fiber slag
Corn particle (water cut is 15 weight %) is placed steeping tank; And the sulfurous acid solution of introducing 0.25 weight % soaks corn; The temperature of soaking is 50 ℃, and the time of immersion is 55 hours, and the introducing amount of the sulfurous acid solution of 0.25 weight % makes liquid level be higher than 10 centimetres of corn particles.
Corn after soaking connected in the degerming mill machine that water is transported to the DTMT-80 type together take off embryo, being milled to average particle diameter through the corn particle of the draining screen in the germ separator is 3 millimeters particle; Afterwards with this particle with the undersized water of dehydration, be transported in the swirler-, to remove the plumule under grinding; Afterwards; To in swirler-, remove the corn particle that grinds plumule down and water is incorporated into together and proceeds grinding in the DTMT-80 type degerming mill machine; Obtain average particle diameter after the grinding and be 1.5 millimeters particle; Afterwards with this particle with the undersized water of dehydration, be delivered in the swirler-, to remove the plumule under grinding.
The corn particle that has removed plumule is incorporated in the impact grinding of LZM685-NA type grinds, obtain soup compound, and said soup compound is incorporated into pressure curved sieve (Yixing starch instrument factory; DZQ50) sieve in; Component after obtaining the zein fiber slag and having removed the zein fiber slag, wherein, the compass screen surface radian of pressure curved sieve is 120 °; The sieve seam width of pressure curved sieve is 75 microns, and feed pressure is 0.2MPa.The component of having removed behind the zein fiber slag is used to prepare milk of starch.
The zein fiber slag that obtains joined in the machine of extracting dewater, the content that obtains zein fiber is 25 weight %, and the content of water is 55 weight %, and starch content is 4 weight %, and protein contnt is the zein fiber slag of 10 weight %, and surplus is an equal amount impurity.
2) cultivation of black mold wheat bran
The zein fiber slag that step (1) is obtained is that 121 ℃, pressure are to obtain substratum behind the high-temperature sterilization 50min under the 0.12MPa in temperature; In substratum, insert aspergillus niger strain (slant strains); With every gram substratum is benchmark, and the access amount of said aspergillus niger strain is 2 * 10
5Individual spore.Culture condition comprises: temperature is 32 ℃, and humidity is 47%, and air flow is 0.1 volume: (volume minute); Incubation time is 9 days; And whenever turned over bran once at a distance from 12 hours, obtain the black mold wheat bran, the spore count that contains in the black mold wheat bran that every gram obtains is as shown in table 1.
3) citric acid fermentation
Adding corn enzymolysis clear liquid, water and urea obtain seed culture medium and (make that total sugar content is 9 weight % in seeding tank; Nitrogenous source (in the nitrogen element) is 0.1 weight %); In temperature is that 121 ℃, pressure are high-temperature sterilization fast cooling to 36 ℃ after 50 minutes under the 0.12MPa, and (inoculum size of black mold wheat bran makes that the access amount of aspergillus niger spore is 2 * 10 in every milliliter of seed culture medium to insert the black mold wheat bran
5Individual).Be 3 at 36 ℃, pH value then, air flow is 0.2 volume: carried out spawn culture 30 hours under the condition of (volume minute), obtain seed culture fluid.Measure and can know through sampling sediments microscope inspection, acid test and pH, the pH of seed culture fluid is 2.0, acidity is 1%, bacterium ball size evenly, mycelia is sturdy stretches out.
Adding corn enzymolysis clear liquid, corn enzymolysis residue, water and urea obtain fermention medium and (make that total sugar content is 15.5 weight % in fermentor tank; Nitrogenous source (in the nitrogen element) is 0.08 weight %); In temperature is that 121 ℃, pressure are high-temperature sterilization fast cooling to 39 ℃ after 50 minutes under the 0.12MPa; Insert step 2) seed culture fluid that obtains, the inoculum size of seed culture fluid makes that the access amount of aspergillus niger spore is 1.5 * 10 in every milliliter of fermention medium
4Individual.And be 1.8 at 35 ℃, pH value, air flow is 0.12 volume: fermented 60 hours under the condition of (volume minute), obtain fermented liquid.The result of the transformation efficiency of fermenting acidity and Hydrocerol A is as shown in table 2.
Embodiment 2
1) preparation of zein fiber slag
Corn particle (water cut is 15 weight %) is placed steeping tank; And the sulfurous acid solution of introducing 0.2 weight % soaks corn; The temperature of soaking is 45 ℃, and the time of immersion is 50 hours, and the introducing amount of the sulfurous acid solution of 0.2 weight % makes liquid level be higher than 10 centimetres of corn particles.
Corn after soaking connected in the degerming mill machine that water is transported to the DTMT-80 type together take off embryo, being milled to average particle diameter through the corn particle of the draining screen in the germ separator is 4 millimeters particle; Afterwards with this particle with the undersized water of dehydration, be transported in the swirler-, to remove the plumule under grinding; Afterwards; To in swirler-, remove the corn particle that grinds plumule down and water is incorporated into together and proceeds grinding in the DTMT-80 type degerming mill machine; Obtain average particle diameter after the grinding and be 1.5 millimeters particle; Afterwards with this particle with the undersized water of dehydration, be delivered in the swirler-, to remove the plumule under grinding.
The corn particle that has removed plumule is incorporated in the impact grinding of LZM685-NA type grinds, obtain soup compound, and said soup compound is incorporated into pressure curved sieve (Yixing starch instrument factory; DZQ50) sieve in; Component after obtaining the zein fiber slag and having removed the zein fiber slag, wherein, the compass screen surface radian of pressure curved sieve is 120 °; The sieve seam width of pressure curved sieve is 75 microns, and feed pressure is 0.3MPa.The component of having removed behind the zein fiber slag is used to prepare milk of starch.
The zein fiber slag that obtains joined in the machine of extracting dewater, the content that obtains zein fiber is 30 weight %, and the content of water is 55 weight %, and starch content is 2 weight %, and protein contnt is the zein fiber slag of 7 weight %, and surplus is an equal amount impurity.
2) cultivation of black mold wheat bran
The zein fiber slag that step (1) is obtained is being that 121 ℃, pressure are to obtain substratum behind the high-temperature sterilization 50min under the 0.12MPa in temperature; In substratum, insert aspergillus niger strain (slant strains); With every gram substratum is benchmark, and the access amount of said aspergillus niger strain is 4 * 10
5Individual spore.Culture condition comprises: temperature is 35 ℃, and humidity is 53%, and air flow is 0.2 volume: (volume minute); Incubation time is 11 days; And whenever turned over bran once at a distance from 12 hours, obtain the black mold wheat bran, the spore count that contains in the black mold wheat bran that every gram obtains is as shown in table 1.
3) citric acid fermentation
Adding corn enzymolysis clear liquid, water and urea obtain seed culture medium and (make that total sugar content is 11 weight % in seeding tank; Nitrogenous source (in the nitrogen element) is 0.3 weight %); In temperature is that 121 ℃, pressure are high-temperature sterilization fast cooling to 36 ℃ after 50 minutes under the 0.12MPa, and (inoculum size of black mold wheat bran makes that the access amount of aspergillus niger spore is 3 * 10 in every milliliter of seed culture medium to insert the black mold wheat bran
5Individual).Be 2.5 at 38 ℃, pH value then, air flow is 0.4 volume: carried out spawn culture 22 hours under the condition of (volume minute), obtain seed culture fluid.Measure and can know through sampling sediments microscope inspection, acid test and pH, the pH of seed culture fluid is 2.1, acidity is 1.2%, bacterium ball size evenly, mycelia is sturdy stretches out.
Adding corn enzymolysis clear liquid, corn enzymolysis residue, water and urea obtain fermention medium and (make that total sugar content is 17 weight % in fermentor tank; Nitrogenous source (in the nitrogen element) is 0.15 weight %); In temperature is that 121 ℃, pressure are high-temperature sterilization fast cooling to 38 ℃ after 50 minutes under the 0.12MPa; Insert step 2) seed culture fluid that obtains, the inoculum size of seed culture fluid makes that the access amount of aspergillus niger spore is 3.5 * 10 in every milliliter of fermention medium
4Individual.And be 0.3 volume: fermented 60 hours under the condition of (volume minute), obtain fermented liquid at 34 ℃, pH value 2.5, air flow.The result of the transformation efficiency of fermenting acidity and Hydrocerol A is as shown in table 2.
Embodiment 3
1) preparation of zein fiber slag
Corn particle (water cut is 15 weight %) is placed steeping tank; And the sulfurous acid solution of introducing 0.25 weight % soaks corn; The temperature of soaking is 50 ℃, and the time of immersion is 55 hours, and the introducing amount of the sulfurous acid solution of 0.25 weight % makes liquid level be higher than 10 centimetres of corn particles.
Corn after soaking connected in the degerming mill machine that water is transported to the DTMT-80 type together take off embryo, being milled to average particle diameter through the corn particle of the draining screen in the germ separator is 3 millimeters particle; Afterwards with this particle with the undersized water of dehydration, be transported in the swirler-, to remove the plumule under grinding; Afterwards; To in swirler-, remove the corn particle that grinds plumule down and water is incorporated into together and proceeds grinding in the DTMT-80 type degerming mill machine; Obtain average particle diameter after the grinding and be 1.5 millimeters particle; Afterwards with this particle with the undersized water of dehydration, be delivered in the swirler-, to remove the plumule under grinding.
The corn particle that has removed plumule is incorporated in the impact grinding of LZM685-NA type grinds, obtain soup compound, and said soup compound is incorporated into pressure curved sieve (Yixing starch instrument factory; DZQ50) sieve in; Component after obtaining the zein fiber slag and having removed the zein fiber slag, wherein, the compass screen surface radian of pressure curved sieve is 120 °; The sieve seam width of pressure curved sieve is 75 microns, and feed pressure is 0.3MPa.The component of having removed behind the zein fiber slag is used to prepare milk of starch.
The zein fiber slag that obtains joined in the machine of extracting dewater, the content that obtains zein fiber is 29 weight %, and the content of water is 53 weight %, and starch content is 3 weight %, and protein contnt is the zein fiber slag of 9 weight %, and surplus is an equal amount impurity.
2) cultivation of black mold wheat bran
The zein fiber slag that step (1) is obtained is being that 121 ℃, pressure are to obtain substratum behind the high-temperature sterilization 50min under the 0.12MPa in temperature; In substratum, insert aspergillus niger strain (slant strains); With every gram substratum is benchmark, and the access amount of said aspergillus niger strain is 3 * 10
5Individual spore.Culture condition comprises: temperature is 34 ℃, and humidity is 50%, and air flow is 0.15 volume: (volume minute), incubation time is 10 days, and whenever turns over bran once at a distance from 12 hours.Obtain the black mold wheat bran, the spore count that contains in the black mold wheat bran that every gram obtains is as shown in table 1.
3) citric acid fermentation
Adding corn enzymolysis clear liquid, water and urea obtain seed culture medium and (make that total sugar content is 10 weight % in seeding tank; Nitrogenous source (in the nitrogen element) is 0.2 weight %); In temperature is that 121 ℃, pressure are high-temperature sterilization fast cooling to 36 ℃ after 50 minutes under the 0.12MPa, and (inoculum size of black mold wheat bran makes that the access amount of aspergillus niger spore is 2.5 * 10 in every milliliter of seed culture medium to insert the black mold wheat bran
5Individual).Be 4.5 at 36 ℃, pH value then, air flow is 0.3 volume: carried out spawn culture 25 hours under the condition of (volume minute), obtain seed culture fluid.Measure and can know through sampling sediments microscope inspection, acid test and pH, the pH of seed culture fluid is 2.3, acidity is 1.3%, bacterium ball size evenly, mycelia is sturdy stretches out.
Adding corn enzymolysis clear liquid, corn enzymolysis residue, water and urea obtain fermention medium and (make that total sugar content is 16.5 weight % in fermentor tank; Nitrogenous source (in the nitrogen element) is 0.13 weight %); In temperature is that 121 ℃, pressure are high-temperature sterilization fast cooling to 38 ℃ after 50 minutes under the 0.12MPa; Insert step 2) seed culture fluid that obtains, the inoculum size of seed culture fluid makes that the access amount of aspergillus niger spore is 2.5 * 10 in every milliliter of fermention medium
4Individual.And be 2.2 at 36.5 ℃, pH value, air flow is 0.2 volume: send out 50 hours under the condition of (volume minute), obtain fermented liquid.The result of the transformation efficiency of fermenting acidity and Hydrocerol A is as shown in table 2.
Embodiment 4
1) preparation of zein fiber slag
Corn particle (water cut is 15 weight %) is placed steeping tank; And the sulfurous acid solution of introducing 0.15 weight % soaks corn; The temperature of soaking is 60 ℃, and the time of immersion is 55 hours, and the introducing amount of the sulfurous acid solution of 0.15 weight % makes liquid level be higher than 10 centimetres of corn particles.
Corn after soaking connected in the degerming mill machine that water is transported to the DTMT-80 type together take off embryo, being milled to average particle diameter through the corn particle of the draining screen in the germ separator is 5 millimeters particle; Afterwards with this particle with the undersized water of dehydration, be transported in the swirler-, to remove the plumule under grinding; Afterwards; To in swirler-, remove the corn particle that grinds plumule down and water is incorporated into together and proceeds grinding in the DTMT-80 type degerming mill machine; Obtain average particle diameter after the grinding and be 1.5 millimeters particle; Afterwards with this particle with the undersized water of dehydration, be delivered in the swirler-, to remove the plumule under grinding.
The corn particle that has removed plumule is incorporated in the impact grinding of LZM685-NA type grinds, obtain soup compound, and said soup compound is incorporated into pressure curved sieve (Yixing starch instrument factory; DZQ50) sieve in; Component after obtaining the zein fiber slag and having removed the zein fiber slag, wherein, the compass screen surface radian of pressure curved sieve is 120 °; The sieve seam width of pressure curved sieve is 75 microns, and feed pressure is 0.25MPa.The component of having removed behind the zein fiber slag is used to prepare milk of starch.
The zein fiber slag that obtains joined in the machine of extracting dewater, the content that obtains zein fiber is 33 weight %, and the content of water is 45 weight %, and starch content is 5 weight %, and protein contnt is the zein fiber slag of 11 weight %, and surplus is an equal amount impurity.
2) cultivation of black mold wheat bran
According to the step 2 among the embodiment 1) method carry out the cultivation of black mold wheat bran, different is: substratum is an above-mentioned steps 1) in the zein fiber slag that obtains, obtain the black mold wheat bran, the spore count that contains in the black mold wheat bran that every gram obtains is as shown in table 1.
3) citric acid fermentation
Method according to the step 3) among the embodiment 1 is carried out citric acid fermentation, and different is, and what to insert seeding tank is above-mentioned steps 2) in the black mold wheat bran that obtains, the result of the fermenting acidity of the fermented liquid that obtains and the transformation efficiency of Hydrocerol A is as shown in table 2.
Comparative Examples 1
1) preparation of Testa Tritici
Through after the removal of impurities, the water wheat wetting makes moisture content even with wheat; Reach 14-15 weight %; Sieve through mechanical mill then, and then hit wheat bran and friction through bran finisher is powerful, separate adherent powder on the bran flakes with wheat bran; The process powder coater is under the brush of the powder on the bran flakes again, and the process rotary strainer is handled at last, repeatedly elutriation obtains Testa Tritici.The content of fiber is 25 weight % in the Testa Tritici that obtains, and the content of water is 55 weight %, and starch content is 4 weight %, and protein contnt is 9 weight %, and surplus is an equal amount impurity.
2) cultivation of black mold wheat bran
According to the step 2 among the embodiment 1) method carry out the cultivation of black mold wheat bran, different is that substratum is above-mentioned Testa Tritici.Obtain the black mold wheat bran, the spore count that every gram obtains containing in the black mold wheat bran is as shown in table 1.
3) citric acid fermentation
Method according to the step 3) among the embodiment 1 is carried out citric acid fermentation, and different is, and what to insert seeding tank is above-mentioned steps 2) in the black mold wheat bran that obtains.The result of the transformation efficiency of fermenting acidity and Hydrocerol A is as shown in table 2.
Table 1
| Numbering | Every gram obtains the spore count (hundred million) that contains in the black mold wheat bran |
| Embodiment 1 | 14.85 |
| Embodiment 2 | 14.83 |
| Embodiment 3 | 14.81 |
| Embodiment 4 | 14.45 |
| Comparative Examples 1 | 14.22 |
Table 2
| Numbering | Fermentation conversion rate (%) | Ferment strength (kg/ (m 3h)） | Fermentation grain consumption (t) |
| Embodiment 1 | 97.3 | 2.379 | 1.552 |
| Embodiment 2 | 97.4 | 2.384 | 1.558 |
| Embodiment 3 | 97.1 | 2.391 | 1.563 |
| Embodiment 4 | 96.3 | 2.256 | 1.565 |
| Comparative Examples 1 | 94.9 | 2.102 | 1.579 |
Can find out the black mold wheat bran that uses the aspergillus niger spore number in the black mold wheat bran that the cultural method of black mold wheat bran of the present invention obtains to obtain in the Comparative Examples 1 through table 1 and table 2.In addition; The black mold wheat bran that uses the cultural method of black mold wheat bran of the present invention to obtain carries out transformation efficiency and ferment strength that inoculation fermentation prepares Hydrocerol A apparently higher than the method for using the black mold wheat bran that obtains in the Comparative Examples 1 to ferment, and fermentation grain consumption is less than the method for using the black mold wheat bran that obtains in the Comparative Examples 1 to ferment.
Claims (10)
1. the cultural method of a black mold wheat bran, this method comprises sterilizes substratum, in culture medium after sterilization, inserts aspergillus niger strain and carries out wheat bran and cultivate, and it is characterized in that said substratum contains the zein fiber slag.
2. cultural method according to claim 1, wherein, in the said zein fiber slag, the content of zein fiber is 20-35 weight %, and the content of water is 45-65 weight %, and contents of starch is 1.5-5 weight %, and Protein content is 5-15 weight %.
3. cultural method according to claim 2, wherein, in the said zein fiber slag, the content of zein fiber is 25-30 weight %, and the content of water is 54-60 weight %, and contents of starch is 2-4 weight %, and Protein content is 7-12 weight %.
4. cultural method according to claim 1 wherein, is a benchmark with every gram substratum, and the access amount of said aspergillus niger strain is 1.5 * 10
5~ 5.5 * 10
5Individual aspergillus niger spore.
5. cultural method according to claim 1, wherein, carry out the wheat bran culture condition and comprise: temperature is 30-37 ℃, and humidity is 45-55%, and air flow is the 0.1-0.3 volume: (volume minute), incubation time is 7-15 days.
6. fermentative prepn methods of citric acid; It is characterized in that; This method comprise with the resulting black mold wheat bran of cultural method that adopts any described black mold wheat bran among the claim 1-5 be linked into carry out seed culture in the Hydrocerol A seed culture medium after, the seed culture fluid that obtains is linked into carries out fermentation culture in the citric acid fermentation substratum again.
7. method according to claim 6 wherein, is a benchmark with every milliliter of Hydrocerol A seed culture medium, and the access amount of black mold wheat bran makes that the access amount of aspergillus niger spore is 1.5 * 10
5~ 4.5 * 10
5Individual.
8. according to claim 6 or 7 described methods, wherein, the condition of said seed culture comprises: the temperature of cultivation is 30-42 ℃, and the pH value is 2-6, and air flow is the 0.1-0.5 volume: (volume minute), the time of cultivation is 18-40 hour.
9. method according to claim 6 wherein, is a benchmark with every milliliter of citric acid fermentation substratum, and the access amount of seed culture fluid makes that the access amount of aspergillus niger spore is 1 * 10
4~ 5 * 10
4Individual.
10. according to claim 6 or 8 described methods, wherein, the condition of citric acid fermentation comprises: the temperature of cultivation is 33-40 ℃, and the pH value is 1.5-6, and air flow is the 0.1-0.5 volume: (volume minute), the time of cultivation is 45-65 hour.
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| CN111349570A (en) * | 2018-12-24 | 2020-06-30 | 中粮生物化学（安徽）股份有限公司 | Method for preparing aspergillus niger seeds and fermentation method of citric acid |
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| CN113621522B (en) * | 2021-07-20 | 2023-11-03 | 佛山市海天(高明)调味食品有限公司 | Mould fermentation medium and application thereof |
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