CN104164458A - Processing method of starch raw material and method for preparing citric acid - Google Patents
Processing method of starch raw material and method for preparing citric acid Download PDFInfo
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- CN104164458A CN104164458A CN201410367218.0A CN201410367218A CN104164458A CN 104164458 A CN104164458 A CN 104164458A CN 201410367218 A CN201410367218 A CN 201410367218A CN 104164458 A CN104164458 A CN 104164458A
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Abstract
The invention discloses a processing method of a starch raw material, wherein the method includes the starch raw material, mycelia and water are mixed and subjected to slurry conditioning to obtain a starch slurry, and the starch slurry, an amylase and a protease are mixed and subjected to enzymolysis, wherein the mycelia are obtained after solid liquid separation of a citric acid fermentation liquid; relative to 100 parts by weight of the starch raw material, the mycelia amount is 5-40 parts by weight, and the water amount is 50-500 parts by weight. The invention also provides a method for preparing citric acid through using the enzymatic hydrolysate obtained by the processing method. The processing method of the starch raw material can reduce grain consumption, and the protein content of the obtained enzymatic hydrolysate is high. Moreover, the obtained enzymatic hydrolysate is adopted to be subjected to citric acid fermentation, and the conversion rate of citric acid fermentation can be increased.
Description
Technical field
The present invention relates to a kind for the treatment of process of starchy material and the preparation method of citric acid.
Background technology
Citric acid is a kind of organic acid that is widely used in the industries such as beverage, food and medicine.At present, citric acid is mainly prepared by fermentation method, for example, generally need first starchy material to be pulverized, product and water after pulverizing are mixed to get to farinaceous size, farinaceous size is mixed and carries out enzymolysis with enzyme, obtain enzymolysis product (enzymatic liquefaction liquid), and aspergillus niger is seeded to fermentation generation citric acid in the fermented liquid that contains described enzymolysis product.
In above-mentioned technique, the grain of citric acid fermentation consumption is higher, and the protein content in enzymolysis solution and the transformation efficiency that uses this enzymolysis solution to carry out citric acid fermentation need to improve.
Summary of the invention
The object of the present invention is to provide a kind for the treatment of process of new starchy material and by using the enzymolysis solution that this treatment process obtains to carry out method prepared by citric acid.By the treatment process of starchy material of the present invention, can reduce grain consumption, and protein content in the enzymolysis solution obtaining is high.And carry out citric acid fermentation by the enzymolysis solution obtaining described in using, can improve the transformation efficiency of citric acid fermentation.
The present inventor finds by deep research, by the mycelium obtaining after citric acid fermentation broth solid-liquid separation is prepared to enzymolysis solution together with starchy material, can significantly improve the protein content of enzymolysis solution, and can be effectively utilized by aspergillus niger, thereby promote thalli growth, improve thalline vigor, shorten fermentation period, and improve the transformation efficiency of citric acid fermentation, thereby complete the present invention.
To achieve these goals, the invention provides a kind for the treatment of process of starchy material, wherein, the method comprises: starchy material is mixed to size mixing obtaining farinaceous size with mycelium and water, farinaceous size is mixed and carries out enzymolysis with amylase and proteolytic enzyme; Wherein, described mycelium is the mycelium obtaining after citric acid fermentation broth solid-liquid separation; With respect to the starchy material of 100 weight parts, described mycelial consumption is 5-40 weight part, and the consumption of described water is 50-500 weight part.
By method of the present invention, can significantly improve the protein content of enzymolysis solution, and the enzymolysis solution obtaining is easy to be effectively utilized by aspergillus niger, thereby promotes thalli growth, improve thalline vigor, shorten fermentation period, and improve the transformation efficiency of citric acid fermentation.
Other features and advantages of the present invention are described in detail the embodiment part subsequently.
Embodiment
Below the specific embodiment of the present invention is elaborated.Should be understood that, embodiment described herein only, for description and interpretation the present invention, is not limited to the present invention.
The treatment process of starchy material of the present invention comprises: starchy material is mixed to size mixing obtaining farinaceous size with mycelium and water, farinaceous size is mixed and carries out enzymolysis with amylase and proteolytic enzyme; Wherein, described mycelium is the mycelium obtaining after citric acid fermentation broth solid-liquid separation; With respect to the starchy material of 100 weight parts, described mycelial consumption is 5-40 weight part, and the consumption of described water is 50-500 weight part.
According to the present invention, described mycelium is the mycelium obtaining after citric acid fermentation broth solid-liquid separation.In preferred situation, described mycelium is the mycelium obtaining after citric acid fermentation broth solid-liquid separation.The method of described solid-liquid separation can adopt the conventional the whole bag of tricks using in this area, for example filter or centrifugal etc., there is no particular limitation for the condition of solid-liquid separation, as long as can obtain the mycelium of following water-content, those skilled in the art can suitably select.
According to the present invention, although in the present invention, be that 0 % by weight also can realize the present invention even if make described mycelial water-content, but, for the ease of operating, reducing costs, preferably described mycelial water-content is more than 60 % by weight, more preferably more than 65 % by weight.In addition, preferably described mycelial water-content is below 95 % by weight, more preferably, below 90 % by weight, more preferably, below 80 % by weight, is further preferably below 70 % by weight.By making described mycelial water-content in above-mentioned scope, have advantages of that farinaceous size mixes, is convenient to enzymolysis.
According to the present invention, the solid content of described farinaceous size can be 20-45 % by weight, for farinaceous size good fluidity, be convenient to carry, enzymolysis is simultaneously more thorough, and ensures the sugared sugared concentration of fermentation, and the solid content of preferred described farinaceous size is 30-40 % by weight.By making the solid content of described farinaceous size in above-mentioned scope, have advantages of that enzymolysis solution sugar concentration stabilize, enzymolysis solution protein content are more conducive to citric acid fermentation.
According to the present invention, in order to obtain the farinaceous size of above-mentioned solid content, and in order to improve the protein content of enzymolysis solution, under preferable case, with respect to the starchy material of 100 weight parts, described mycelial consumption is 8-40 weight part, and the consumption of described water is 100-480 weight part; More preferably, with respect to the starchy material of 100 weight parts, described mycelial consumption is 10-30 weight part, and the consumption of described water is 200-450 weight part.
According to the present invention, described starchy material is mixed to the condition of sizing mixing with mycelium and water and comprise: temperature is 60-70 DEG C, and pH value is controlled at 5.2-5.8; Under preferable case, described starchy material is mixed to the condition of sizing mixing with mycelium and water and comprise: temperature is 65-70 DEG C, and pH value is controlled at 5.4-5.7.
According to the present invention, described starchy material is selected from least one in corn, potato class, wheat and Chinese sorghum.After preferably starchy material being pulverized, use.Condition and mode that starchy material is pulverized have no particular limits, as long as can make starchy material fully broken, under preferable case, the particle diameter of the product after the pulverizing obtaining is 50-1000 micron, is preferably 300-600 micron.
According to the present invention, described farinaceous size is mixed with amylase and proteolytic enzyme and carry out the method for enzymolysis there is no particular limitation, can be the conventional method using in this area.But preferably described farinaceous size is mixed to the method for carrying out enzymolysis with amylase and proteolytic enzyme be: described farinaceous size is mixed with partial starch enzyme, obtain mixture, this mixture is contacted with steam, the temperature of the mixture after the condition of contact makes to contact with steam is 85-105 DEG C, and keep 90-180 minute at this temperature, by obtain contact with steam after mixture carry out flash distillation, then, under enzymatic hydrolysis condition, by the product after flash distillation with another part amylase and proteolytic enzyme mixes and carry out enzymolysis.
Wherein, the 60-80 % by weight that the consumption of described partial starch enzyme is diastatic total consumption, the 20-40 % by weight that the diastatic consumption of described another part is diastatic total consumption; Preferably, the 70-76 % by weight that the consumption of described partial starch enzyme is diastatic total consumption, the 24-30 % by weight that the diastatic consumption of described another part is diastatic total consumption.
According to the present invention, to farinaceous size and partial starch enzyme mix that there is no particular limitation, be conducive to the mixing of the two and follow-up gelatinization but temperature is slightly high, preferably, the temperature of mixing is 50-60 DEG C.Farinaceous size is wider with the selectable range of the condition that the mixture of partial starch enzyme contacts with steam, as long as ensure that the rear farinaceous size of contact and diastatic mixture can reach respectively its suitable temperature, for example, the condition that described mixture contacts with steam comprises usage ratio and the duration of contact etc. of temperature, steam and the mixture of contact.Under preferable case, the condition of contact comprises that the temperature of steam is 150-240 DEG C, and the weight ratio of steam and mixture is 0.1-0.2:1, and the time of contact is 3-10 second; More preferably in situation, consider the energy and cost, the temperature of steam is 150-170 DEG C, and the weight ratio of steam and mixture is 0.13-0.17:1, and the time of contact is 3-6 second.
Wherein, farinaceous size is not particularly limited with the mode that diastatic mixture contacts with steam, under preferable case, can for example, at injector known in those skilled in the art (, million smooth injectors or day long water heater.) in spray contact, that is, farinaceous size and diastatic mixture and steam are ejected into simultaneously, fast and in injector, carry out moment and contact, and the mixture after this contact is sent into and in another hold-up vessel, is incubated at a certain temperature processing.
According to the present invention, in order to reach better more abundant by the expansion that makes starch molecule that temperature variation is brought fast, to such an extent as to can make the object that macrobead starch swelling fracture is starch granule, and make the better effects if that contacts of amylase and starch granules, to reach better hydrolysis result, the present invention adopts the mode of flash distillation that the mixture after contacting with steam is cooled to 50-80 DEG C (being preferably 55-65 DEG C).Wherein, flash distillation refers to become saturated steam and the saturation water under a part of container pressure because the unexpected reduction of pressure makes these saturation waters after the saturation water of high pressure enters in the container of relatively low pressure.
According to the present invention, the selectable range of the condition of described flash distillation is wider, as long as reach the object that makes it cooling after flash distillation, for example, the mode of described flash distillation can be atmospheric flashing, can be also vacuum flashing (vacuum flash), and the present invention preferably uses the method for vacuum flashing, the condition of described flash distillation comprises that the vacuum tightness of flash distillation can be-0.05 to-0.2MPa (gauge pressure), and the time of flash distillation can be 5-20 second.
According to the present invention, the consumption of described amylase and proteolytic enzyme can change in the larger context, in preferred situation, in starchy material and mycelial dry weight 1g, described diastatic consumption is 5-20 enzyme activity unit, and the consumption of described proteolytic enzyme is 3-8 enzyme activity unit; In preferred situation, in starchy material and mycelial dry weight 1g, described diastatic consumption is 10-16 enzyme activity unit, and the consumption of described proteolytic enzyme is 4-6 enzyme activity unit.
In the present invention, described diastatic enzyme activity unit is defined as: be 6.0 in pH value, temperature is that under the condition of 70 DEG C, it is an enzyme activity unit that 1 milligram of starch is converted into the required enzyme amount of reducing sugar by 1 minute.The enzyme activity unit of described proteolytic enzyme is defined as: 1g solid enzyme powder, under 40 DEG C of (pH=7.5) conditions, it is an enzyme activity unit that 1min hydrolyzed casein produces 1 μ g tyrosine.
In the present invention, amylase refers to the general name of class of enzymes that can starch-splitting glycosidic link, and described amylase generally comprises α-amylase, beta-amylase, saccharifying enzyme and isoamylase.
α-amylase claims again starch Isosorbide-5-Nitrae-dextrinase, and it can cut the α-Isosorbide-5-Nitrae-glycosidic link of starch chain inside at random, brokenly, by Starch Hydrolysis be maltose, the oligosaccharides that contains 6 glucose units and the oligosaccharides with side chain.The microorganism that produces this enzyme mainly has Bacillus subtilus, aspergillus niger, aspergillus oryzae and head mold.
Beta-amylase claims again starch Isosorbide-5-Nitrae-maltoside enzyme, can cut Isosorbide-5-Nitrae-glycosidic link from starch molecule non reducing end, generates maltose.The product that this enzyme acts on starch is maltose and limit dextrin.This enzyme is mainly produced by aspergillus, head mold and endomyces.
Saccharifying enzyme claims again starch α-Isosorbide-5-Nitrae-glucuroide, and this enzyme acts on the non reducing end of starch molecule, taking glucose as unit, acts on successively the α-Isosorbide-5-Nitrae-glycosidic link in starch molecule, generates glucose.Saccharifying enzyme acts on that product after amylopectin has glucose and with α-1, the oligosaccharides of 6-glycosidic link; The product acting on after amylose starch is all almost glucose.It is mainly aspergillus niger (left U.S. aspergillus, Aspergillus awamori), head mold (snow-white enzyme, De Shi head mold), endomycopsi.sp, monascus that this enzyme produces bacterium.
Isoamylase claims again starch α-1,6-glucuroide, branching enzyme, and this enzyme acts on α-1 at amylopectin molecule branching-point place, and 6-glycosidic link cuts the whole side chain of amylopectin to become amylose starch.It is mainly to dislike the bacteriums such as gas bacillus, genus bacillus and some Pseudomonas that this enzyme produces bacterium.
According to the present invention, preferably use α-amylase and/or isoamylase.More preferably purchased from the Jie Neng section amylase of Wuxi biotechnology company limited of Jie Neng section.
In the present invention, described proteolytic enzyme generally comprises aspartic protease, neutral protease, Sumizyme MP.Be preferably aspartic protease and neutral protease.Described proteolytic enzyme for example can use Jie Neng section thermostable proteinase or Novi's letter proteolytic enzyme.
According to the present invention, described enzymatic hydrolysis condition comprises: hydrolysis temperature is 50-80 DEG C, and pH value is 5-6, and enzymolysis time is 10-60 minute; Preferably hydrolysis temperature is 55-65 DEG C, and pH value is 5.2-5.8, and enzymolysis time is 15-40 minute.
After enzymolysis, carry out the effect of iodine examination detection enzymolysis, after iodine examination qualified (orange or redness), part enzymatic liquefaction liquid can be used as fermentation bed material and cultivates seed, major part enters plate-and-frame filter press and carries out press filtration, realize solid-liquid separation, obtain enzymolysis (liquefaction) clear liquid and enzymolysis (liquefaction) residue, prepare citric acid for fermentation.
Thus, the present invention also provides a kind of preparation method of citric acid, and the method comprises according to above-mentioned treatment process to be processed starchy material, obtains enzymolysis product, and the fermented liquid that contains this enzymolysis product is fermented under aspergillus niger exists.
According to the present invention, in described fermented liquid, the content of each component can in very large range change, under preferable case, can also in described fermented liquid, add supplementary nitrogenous source as required, the content of described supplementary nitrogenous source can be the 0.05-0.2 % by weight of fermented liquid gross weight.According to the present invention, the kind of described supplementary nitrogenous source is conventionally known to one of skill in the art, and for example, described supplementary nitrogenous source can be at least one in urea, ammonium sulfate and ammonium nitrate.In addition, can also in fermented liquid, supplement appropriate water according to the requirement of fermented liquid liquid level, the selectable range of the amount of water is wider, can be decided according to the actual requirements.
According to the present invention, the enzymolysis product that described fermented liquid contains starchy material, the enzymolysis product of described starchy material contains starchy material enzymolysis residue and starchy material enzymolysis clear liquid, in order to be easier to the carrying out of fermentation, taking the gross weight of the enzymolysis product of the starchy material that contains in fermented liquid as benchmark, the content of described starchy material enzymolysis clear liquid is 80-95 % by weight, and the content of described starchy material enzymolysis residue is 5-20 % by weight.
According to the present invention, the inoculum size of described aspergillus niger can in very large range change, and under preferable case, taking every gram of fermented liquid as benchmark, the inoculum size of aspergillus niger is 1 × 10
4-5 × 10
4individual colony-forming unit (spore), more preferably 2 × 10
4-4 × 10
4individual colony-forming unit.The condition of described fermentation has no particular limits, for example, can comprise: temperature is 30-40 DEG C, and air flow is 0.1-1 volume: volume minute, the time of fermentation is 50-60 hour.
Described colony-forming unit is defined as a certain amount of bacterium liquid after dilution by the method for pouring into a mould or being coated with, and allows unicellular being dispersed in one by one on culture medium flat plate of microorganism in it, and after cultivating, each viable cell just forms a bacterium colony.It is the single celled number containing in every milliliter of bacterium liquid.
Described colony-forming unit can be measured by means commonly known in the art, for example, counts by blood counting chamber.
Term " air flow " generally with ventilation recently represents, conventionally recently to represent (V/Vmin) by the volume of air of unit volume nutrient solution in per minute, for example ventilation ratio is 1:0.1-1, abbreviation air flow be 0.1-1 volume: volume minute.
The ferment aspergillus niger that uses of the present invention can be aspergillus niger solid preparation or the aspergillus niger strain being purchased, for example, and aspergillus niger Co827 (Shanghai Industrial institute of microbiology) or aspergillus niger T01 (Tianjin industrial microorganism institute).
Described aspergillus niger can adopt conventional method inoculation, for example, and before being seeded in fermented liquid, by the processing of described aspergillus niger process seed culture, be seeded in aspergillus niger nutrient solution by aspergillus niger, obtain aspergillus niger seed liquor, afterwards the seed liquor obtaining is joined in fermented liquid.The degree of aspergillus niger seed culture can be measured and be observed the growth of aspergillus niger by sampling sediments microscope inspection, acid test and pH, when pH 2.0-2.5, acidity 0.5-2.0%, bacterium ball size evenly, mycelia is sturdy stops cultivating while stretching out.
According to the present invention, the preparation method of described aspergillus niger nutrient solution has no particular limits, as long as the nutrient solution obtaining can be applicable to the cultivation of aspergillus niger, for example, can, by the enzymolysis product sterilizing obtaining according to method of the present invention, obtain nutrient solution.
In the present invention, in described aspergillus niger seed culture treating processes, the inoculum size of aspergillus niger can in very large range change, and under preferable case, taking every gram of aspergillus niger nutrient solution as benchmark, the inoculum size of aspergillus niger is 2 × 10
5-4 × 10
5individual colony-forming unit.
According to the present invention, the condition that described fermentation of Aspergillus niger is cultivated can in very large range change, the condition of for example described cultivation can comprise: the temperature of cultivation can be 25-40 DEG C, pH value can be 2-7, air flow can be 0.1-1 volume: volume minute, and the time of cultivation can be 20-40 hour; In preferred situation, the condition of described cultivation comprises: the temperature of cultivation is 30-40 DEG C, and pH value is 2.5-6.5, and air flow is 0.2-0.8 volume: volume minute, the time of described cultivation is 25-35 hour.
Tunning citric acid can, by conventional method, separate and refine according to the requirement of different Industrial products, such as neutralization, acidolysis, decolouring, concentrated, crystallization, packaging.
Below in conjunction with embodiment, the present invention will be described in more detail.
In the embodiment of the present invention, amylase used is the Jie Neng section amylase purchased from Wuxi biotechnology company limited of Jie Neng section; Proteolytic enzyme used is the Jie Neng section thermostable proteinase purchased from Wuxi biotechnology company limited of Jie Neng section; Aspergillus niger T01 used is purchased from Tianjin industrial microorganism institute; Injector used is million smooth injectors.
In following examples, the mycelium that the mycelium using obtains after for citric acid fermentation broth solid-liquid separation.
The measuring method of the moisture in mycelium is: take the mycelium of W1 weight, dry to constant weight W2, moisture content=the 1-W2/W1 in mycelium in 105 DEG C of baking ovens.In addition, mycelial dry weight is W2.
According to the concentration (abbreviation acidity) of GB 1987-2007 standard detection fermentation secondary fermentation liquid, and calculate the transformation efficiency of citric acid, weight × 100% of the volume/total reducing sugar (total reducing sugar=seeding tank total reducing sugar+fermentor tank total reducing sugar) of concentration (abbreviation the acidity) × fermented liquid of transformation efficiency (%)=fermented liquid.
In following examples, the protein content in enzymolysis product is measured by the method for Micro-kjoldahl method.
Embodiment 1
1) 62 tons of corns are pulverized, obtained average particle diameter and be the crushed products of 400 microns.By the product after pulverizing and mycelium (water content is 60 % by weight) and the 254.2m of 6.2 tons
3under the condition that water in temperature is at 60 DEG C, pH value is 5.2, mix, obtain farinaceous size, wherein, the solid content of farinaceous size is 20 % by weight.
2) by step 1) farinaceous size that obtains and 12.1kg amylase (the total weight 1g that is equivalent to the corn pulverized and mycelium dry weight uses the amylase of 7.5 enzyme activity units) mixes, obtain mixture, this mixture is sprayed in injector with the steam of 150 DEG C and contact (weight ratio of steam and mixture is 0.15:1), the time of contact is 5 seconds, the temperature that makes the mixture after contacting with steam is 105 DEG C, and at this temperature, keeps 90 minutes;
Mixture after above-mentioned steps is contacted with steam carries out flash distillation, and (vacuum tightness is-0.1MPa (gauge pressure), time is 10 seconds) to 55 DEG C, and ((being equivalent to the proteolytic enzyme of 4 enzyme activity units of total weight 1g use of the corn pulverized and mycelium dry weight) mixes with the 4.0Kg amylase amylase of 2.5 enzyme activity units (be equivalent to the corn pulverized and the total weight 1g of mycelium dry weight use) and 5kg proteolytic enzyme, regulate pH value to 5.2, and at this temperature, keep 20 minutes, obtain enzymolysis product (protein content in enzymolysis product is 1.6875 % by weight).
3) the Partial digestion product (80 % by weight of enzymolysis product gross weight) step (2) being obtained, by carrying out press filtration with fluid pressure type sheet frame pressure filter, is isolated enzymolysis clear liquid and enzymolysis solid phase residue (water content is 50% weight).
The enzymolysis product configuration fermented liquid that uses above-mentioned steps to obtain, joins in fermentor tank after specifically consisting of the enzymolysis solid phase residue of enzymolysis clear liquid, 19.9 weight parts of 80 weight parts and 0.1 weight part urea sterilizing, obtains fermented liquid.
Another part enzymolysis product obtained above is heated to 121 DEG C of sterilizations, maintain 20 minutes after fast cooling to 37 DEG C, access aspergillus niger strain, inoculum size is: every gram of enzymolysis product inoculation 3 × 10
5individual colony-forming unit, at 36 DEG C, 0.3 volume: carry out spawn culture under the aeration condition of volume minute; Measure the growth of aspergillus niger observed by sampling sediments microscope inspection, acid test and pH, when pH value 2.5, acidity 1%, bacterium ball size evenly, mycelia is sturdy while stretching out, stops cultivation.
Above-mentioned cultivation aspergillus niger strain is joined in the fermentor tank of the fermented liquid that contains above-mentioned preparation and ferment, and detect the total reducing sugar in fermented liquid, inoculum size is: every gram of fermented liquid inoculation 3 × 10
4individual colony-forming unit stirs, 0.2 volume: under the aeration condition of volume minute, cultivate 58 hours fermentation ends under 37 DEG C, 90 revs/min.
Acidity, transformation efficiency and the fermentation grain consumption of calculating the fermented liquid after fermentation, result is as shown in table 1.
Embodiment 2
1) 62 tons of corns are pulverized, obtained average particle diameter and be the crushed products of 400 microns.By the product after pulverizing and mycelium (water content is 70 % by weight) and the 69.6m of 18.6 tons
3under the condition that water in temperature is at 70 DEG C, pH value is 5.8, mix, obtain farinaceous size, wherein, the solid content of farinaceous size is 45 % by weight.
2) by step 1) farinaceous size that obtains and 18.9kg amylase (the total weight 1g that is equivalent to the corn pulverized and mycelium dry weight uses the amylase of 11.2 enzyme activity units) mixes, obtain mixture, this mixture is sprayed in injector with the steam of 170 DEG C and contact (weight ratio of steam and mixture is 0.15:1), the time of contact is 3 seconds, the temperature that makes the mixture after contacting with steam is 105 DEG C, and at this temperature, keeps 180 minutes;
Mixture after above-mentioned steps is contacted with steam carries out flash distillation, and (vacuum tightness is-0.1MPa (gauge pressure), time is 6 seconds) to 65 DEG C, and ((being equivalent to the proteolytic enzyme of 6 enzyme activity units of total weight 1g use of the corn pulverized and mycelium dry weight) mixes with the 8.1Kg amylase amylase of 4.8 enzyme activity units (be equivalent to the corn pulverized and the total weight 1g of mycelium dry weight use) and 7.5kg proteolytic enzyme, regulate pH value to 5.8, and at this temperature, keep 20 minutes, obtain enzymolysis product (protein content in enzymolysis product is 1.875 % by weight).
3) the Partial digestion product (85 % by weight of enzymolysis product gross weight) step (2) being obtained, by carrying out press filtration with fluid pressure type sheet frame pressure filter, is isolated enzymolysis clear liquid and enzymolysis solid phase residue (water content is 50% weight).
The enzymolysis product configuration fermented liquid that uses above-mentioned steps to obtain, joins in fermentor tank after specifically consisting of the enzymolysis solid phase residue of enzymolysis clear liquid, 9.9 weight parts of 90 weight parts and 0.1 weight part urea sterilizing, obtains fermented liquid.
Another part enzymolysis product obtained above is heated to 121 DEG C of sterilizations, maintain 20 minutes after fast cooling to 37 DEG C, access aspergillus niger strain, inoculum size is: every gram of enzymolysis product inoculation 3 × 10
5individual colony-forming unit, at 36 DEG C, 0.3 volume: carry out spawn culture under the aeration condition of volume minute; Measure the growth of aspergillus niger observed by sampling sediments microscope inspection, acid test and pH, when pH value 2.5, acidity 1%, bacterium ball size evenly, mycelia is sturdy while stretching out, stops cultivation.
Above-mentioned cultivation aspergillus niger strain is joined in the fermentor tank of the fermented liquid that contains above-mentioned preparation and ferment, and detect the total reducing sugar in fermented liquid, inoculum size is: every gram of fermented liquid inoculation 3 × 10
4individual colony-forming unit stirs, 0.2 volume: under the aeration condition of volume minute, cultivate 57 hours fermentation ends under 37 DEG C, 90 revs/min.
Acidity, transformation efficiency and the fermentation grain consumption of calculating the fermented liquid after fermentation, result is as shown in table 1.
Embodiment 3
1) 62 tons of corns are pulverized, obtained average particle diameter and be the crushed products of 400 microns.By the product after pulverizing and mycelium (water content is 65 % by weight) and the 129.7m of 12.4 tons
3under the condition that water in temperature is at 65 DEG C, pH value is 5.5, mix, obtain farinaceous size, wherein, the solid content of farinaceous size is 32.5 % by weight.
2) by step 1) farinaceous size that obtains and 15.6kg amylase (the total weight 1g that is equivalent to the corn pulverized and mycelium dry weight uses the amylase of 9.4 enzyme activity units) mixes, obtain mixture, this mixture is sprayed in injector with the steam of 160 DEG C and contact (weight ratio of steam and mixture is 0.15:1), the time of contact is 6 seconds, the temperature that makes the mixture after contacting with steam is 105 DEG C, and at this temperature, keeps 135 minutes;
Mixture after above-mentioned steps is contacted with steam carries out flash distillation, and (vacuum tightness is-0.1MPa (gauge pressure), time is 10 seconds) to 60 DEG C, and ((being equivalent to the proteolytic enzyme of 5 enzyme activity units of total weight 1g use of the corn pulverized and mycelium dry weight) mixes with the 5.2Kg amylase amylase of 3.1 enzyme activity units (be equivalent to the corn pulverized and the total weight 1g of mycelium dry weight use) and 6.25kg proteolytic enzyme, regulate pH value to 5.5, and at this temperature, keep 20 minutes, obtain enzymolysis product (protein content in enzymolysis product is 1.781 % by weight).
3) the Partial digestion product (85 % by weight of enzymolysis product gross weight) step (2) being obtained, by carrying out press filtration with fluid pressure type sheet frame pressure filter, is isolated enzymolysis clear liquid and enzymolysis solid phase residue (water content is 50% weight).
The enzymolysis product configuration fermented liquid that uses above-mentioned steps to obtain, joins in fermentor tank after specifically consisting of the enzymolysis solid phase residue of enzymolysis clear liquid, 14.9 weight parts of 85 weight parts and 0.1 weight part urea sterilizing, obtains fermented liquid.
Another part enzymolysis product obtained above is heated to 121 DEG C of sterilizations, maintain 20 minutes after fast cooling to 37 DEG C, access aspergillus niger strain, inoculum size is: every gram of enzymolysis product inoculation 3 × 10
5individual colony-forming unit, at 36 DEG C, 0.3 volume: carry out spawn culture under the aeration condition of volume minute; Measure the growth of aspergillus niger observed by sampling sediments microscope inspection, acid test and pH, when pH value 2.5, acidity 1%, bacterium ball size evenly, mycelia is sturdy while stretching out, stops cultivation.
Above-mentioned cultivation aspergillus niger strain is joined in the fermentor tank of the fermented liquid that contains above-mentioned preparation and ferment, and detect the total reducing sugar in fermented liquid, inoculum size is: every gram of fermented liquid inoculation 3 × 10
4individual colony-forming unit stirs, 0.2 volume: under the aeration condition of volume minute, cultivate 59 hours fermentation ends under 37 DEG C, 90 revs/min.
Acidity, transformation efficiency and the fermentation grain consumption of calculating the fermented liquid after fermentation, result is as shown in table 1.
Embodiment 4
Carry out according to the method for embodiment 1, different, mycelial consumption is 21.7 tons.Wherein, the protein content in the enzymolysis product obtaining is 1.625 % by weight, acidity, transformation efficiency and the fermentation grain consumption of the fermented liquid after fermentation, and result is as shown in table 1.
Comparative example 1
Carry out according to the method for embodiment 1, different is that the mycelium of 6.2 tons is replaced with to 6.2 tons of products after crush maize.Wherein, the protein content in the enzymolysis product obtaining is 1.5625 % by weight, acidity, transformation efficiency and the fermentation grain consumption of the fermented liquid after fermentation, and result is as shown in table 1.
Table 1
| Numbering | Acidity (g/ml) | Fermentation period (h) | Transformation efficiency (%) | Fermentation grain consumption (g/1.h) |
| Embodiment 1 | 15.9 | 58 | 96.2 | 1.560 |
| Embodiment 2 | 15.9 | 57 | 96.2 | 1.560 |
| Embodiment 3 | 15.9 | 59 | 96.2 | 1.560 |
| Embodiment 4 | 15.8 | 62 | 95.6 | 1.570 |
| Comparative example 1 | 15.6 | 65 | 94.4 | 1.590 |
More than describe the preferred embodiment of the present invention in detail; but the present invention is not limited to the detail in above-mentioned embodiment, within the scope of technical conceive 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.It should be noted that in addition, each concrete technical characterictic described in above-mentioned embodiment, in reconcilable situation, can combine by any suitable mode, 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 various embodiment of the present invention, as long as it is without prejudice to thought of the present invention, it should be considered as content disclosed in this invention equally.
Claims (12)
1. a treatment process for starchy material, is characterized in that, the method comprises: starchy material is mixed to size mixing obtaining farinaceous size with mycelium and water, farinaceous size is mixed and carries out enzymolysis with amylase and proteolytic enzyme; Wherein, described mycelium is the mycelium obtaining after citric acid fermentation broth solid-liquid separation; With respect to the starchy material of 100 weight parts, described mycelial consumption is 5-40 weight part, and the consumption of described water is 50-500 weight part.
2. treatment process according to claim 1, wherein, described mycelium is the mycelium obtaining after citric acid fermentation broth solid-liquid separation.
3. treatment process according to claim 1 and 2, wherein, the solid content of described farinaceous size is 20-45 % by weight.
4. according to the treatment process described in any one in claim 1-3, wherein, described mycelial water-content is 60-70 % by weight.
5. according to the treatment process described in any one in claim 1-4, wherein, with respect to the starchy material of 100 weight parts, described mycelial consumption is 10-30 weight part, and the consumption of described water is 200-450 weight part.
6. according to the treatment process described in any one in claim 1-4, wherein, described starchy material is mixed to the condition of sizing mixing with mycelium and water and comprise: temperature is 60-70 DEG C, and pH value is controlled at 5.2-5.8.
7. according to the treatment process described in any one in claim 1-4, wherein, in starchy material and mycelial dry weight 1g, described diastatic consumption is 5-20 enzyme activity unit; The consumption of described proteolytic enzyme is 3-8 enzyme activity unit.
8. treatment process according to claim 7, wherein, farinaceous size being mixed to the method for carrying out enzymolysis with amylase and proteolytic enzyme is: described farinaceous size is mixed with partial starch enzyme, obtain mixture, this mixture is contacted with steam, the temperature of the mixture after the condition of contact makes to contact with steam is 85-105 DEG C, and keep 90-180 minute at this temperature, by obtain contact with steam after mixture carry out flash distillation, then, under enzymatic hydrolysis condition, by the product after flash distillation with another part amylase and proteolytic enzyme mixes and carry out enzymolysis.
9. treatment process according to claim 8, wherein, the condition of described contact comprises that the temperature of steam is 150-240 DEG C, and the weight ratio of steam and mixture is 0.1-0.2:1, and the time of contact is 3-10 second; Described enzymatic hydrolysis condition comprises: hydrolysis temperature is 50-80 DEG C, and pH value is 5-6, and enzymolysis time is 10-60 minute.
10. treatment process according to claim 8 or claim 9, wherein, the 60-80 % by weight that the consumption of described partial starch enzyme is diastatic total consumption, the 20-40 % by weight that the diastatic consumption of described another part is diastatic total consumption.
11. treatment processs according to claim 1, wherein, described starchy material is selected from least one in corn, potato class, wheat and Chinese sorghum.
The preparation method of 12. 1 kinds of citric acids, the method comprises according to the treatment process described in any one in claim 1-11 to be processed starchy material, obtains enzymolysis product, and the fermented liquid that contains this enzymolysis product is fermented under aspergillus niger exists.
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| CN109852640A (en) * | 2017-11-30 | 2019-06-07 | 中粮生物化学（安徽）股份有限公司 | Full starch prepares citric acid fermented with seed culture medium and the citric acid fermented method with culture medium and citric acid |
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