CN101372674B - The production method of Rich chromium yeast - Google Patents
The production method of Rich chromium yeast Download PDFInfo
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- CN101372674B CN101372674B CN200710143066.6A CN200710143066A CN101372674B CN 101372674 B CN101372674 B CN 101372674B CN 200710143066 A CN200710143066 A CN 200710143066A CN 101372674 B CN101372674 B CN 101372674B
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Abstract
The invention provides a kind of method being applicable to suitability for industrialized production Rich chromium yeast, comprise the following steps: A) by yeast saccharomyces cerevisiae bacterial classification amplification culture several times, to provide the yeast saccharomyces cerevisiae bacterial classification of aequum; B) being incorporated in a fermentor tank by the yeast saccharomyces cerevisiae bacterial classification of described aequum, is 3.8 ~ 6.8 at pH, and temperature is the condition bottom fermentation of 28 ~ 35 DEG C, and when weight in wet base reaches more than 30g/L, stream adds chromium salt solution, ferments to stop when weight in wet base reaches 110-150g/L; And C) by centrifugation, washing and dry results Rich chromium yeast product.Wherein, described chromic salts is chromic salt, preferred chromium acetate and CrCl
36H
2o, its consumption is, every 100ml fermented liquid uses 0.05 ~ 0.5g chromic salts.Adopt the Rich chromium yeast that method of the present invention is produced, achieve the conversion of inorganic chromium to organic chromium well, thus be that human body and animal benefit chromium provide approach.
Description
Technical field
The industrialized preparing process that the present invention relates to Rich chromium yeast and the Rich chromium yeast obtained by the method.
Background technology
Chromium is the indispensable a kind of trace element of human body and animal, take part in sugar metabolism and the metabolism of fat of body, has much important physiological function.Regular Insulin is glycometabolic core substance, and Regular Insulin plays a role, and chromium must be had again to participate in.Because trivalent chromium, by forming glucose tolerance factor or other organo-chromium compounds, plays a role with Regular Insulin, and embodies its physiological function.。Chromium can increase decomposition and the excretion of cholesterol, and human body lacks chromium can make fat metabolic disturbance, occurs hyperlipidemia, particularly the disease such as hypercholesterolemia and arteriosclerosis, also can cause the disease such as diabetes and cataract.
Yeast is the widest microorganism of human use, and its protein content, up to more than 50%, also containing abundant vitamin B group and several mineral materials, is called as the source of high-quality nutrition.Yeast has the rich chromium ability of height and chemical chromium is converted into the ability of biological chromium, Rich chromium yeast is exactly add chromium element in the process of culturing yeast, by yeast in process of growth to the autonomous absorption of chromium and conversion, chromium more efficiently, is more safely absorbed by the body utilization.
Summary of the invention
The object of this invention is to provide a kind of method being applicable to suitability for industrialized production Rich chromium yeast, comprise the following steps: A) by yeast saccharomyces cerevisiae bacterial classification amplification culture several times, to provide the yeast saccharomyces cerevisiae bacterial classification of aequum; B) being incorporated in a fermentor tank by the yeast saccharomyces cerevisiae bacterial classification of described aequum, is 3.8 ~ 6.8 at pH, and temperature is the condition bottom fermentation of 28 ~ 35 DEG C, and when weight in wet base reaches more than 30g/L, stream adds chromium salt solution, ferments to stop when weight in wet base reaches 110-150g/L; And C) by centrifugation, washing and dry results Rich chromium yeast product.Wherein, described chromic salts is chromic salt, preferred chromium acetate and CrCl
36H
2o, its consumption is, every 100ml fermented liquid uses 0.05 ~ 0.5g chromic salts.Wherein, A) in substratum used be made up of following ingredients: 1) process water; 2) MgSO
4, ZnSO
4, and KH
2pO
4; 3) molasses; 4) N and P donor solution, it contains urea, NH
4h
2pO
4, (NH
4)
2sO
4, and yeast extract paste.
In one embodiment, above-mentioned substratum has following feature: based on described process water Weight computation, described MgSO
4consumption is 0.05% to 0.1%, described ZnSO
4consumption is 0.05% to 0.1%, and described KH
2pO
4consumption be 0.08% to 0.12%; The consumption of described molasses is 75% for calculating standard with the transformation efficiency that molasses change into yeast; Described N and P donor is the solution containing following ingredients: process water, and based on the % urea of 0.4 ~ 0.8, the NH of 0.4 ~ 0.8% of process water weight
4h
2pO
4, 0.8 ~ 1.2% (NH
4)
2sO
4, and 0.2 ~ 0.4% yeast extract paste.
One preferred embodiment in, described initial medium is further containing vitamin H, calcium pantothenate, VITMAIN B1, Lin Suanna Vitamin B2 Sodium Phosphate and vitamin B6.
One preferred embodiment in, add halfcystine when described weight in wet base reaches 40-70g/L in stepb, and with by water, glucose mother liquid, described molasses, MgSO
4with, ZnSO
4the solution carbon source as a supplement of composition, preferably, described solution contains the glucose of 15 ~ 25%, the vattability sugar content of 25 ~ 35% is 29% molasses, 0.01 ~ 0.05%MgSO
4with 0.01% ~ 0.05%ZnSO
4.
In a preferred embodiment, the configuration proportion of the described solution of carbon source is as a supplement: 40wt% glucose solution: molasses: water: MgSO
4: ZnSO
4=500L: 300L: 200L: 30kg: 30kg.
One preferred embodiment in, step B) in yeast saccharomyces cerevisiae bacterial classification used through three amplification culture, wherein, one-level amplify and secondary amplification be 28 ~ 31 DEG C and 4.8 ~ 5.8 pH value under carry out.For the mixed aqueous solution that the substratum that described one-level is amplified and secondary amplifies is containing 8 ~ 12% sucrose, 1 ~ 3% yeast powder, 0.1 ~ 0.5% potassium primary phosphate and 0.1 ~ 0.5% magnesium sulfate.
One preferred embodiment in, three grades of amplifications carry out under being the condition of 4.5 to 5.5 at temperature, the pH of 26 to 32 DEG C, until stop when weight in wet base reaches 35-40g/L, wherein, pressing the Weight computation of substratum for described three grades of substratum amplified, is by the molasses of 25% to 28%; The urea of 0.08% to 0.11%; The primary ammonium phosphate of 0.07% to 0.16%; The yeast powder of 0.05% to 0.08%; The magnesium sulfate of 0.05% to 0.1%; The process water composition of 0.02% to 0.05% zinc sulfate and surplus.
The Rich chromium yeast obtained according to the inventive method, its chromium content is greater than 2000mg/kg, and protein content is greater than 45.0%, vitamins B
1be greater than 43.0mg/kg, VITAMIN reaches B
2for 63.0mg/kg.
Adopt the Rich chromium yeast that method of the present invention is produced, achieve the conversion of inorganic chromium to organic chromium well, improve the specific absorption of chromium in animal body, thus be that human body and animal benefit chromium provide approach.
The raw material mending chromium protective foods and medicine is can be used as according to the Rich chromium yeast that the inventive method obtains; Can be used as the raw material of chromium nutrition element strengthening in nourishing reinforced food; Can be used for livestock breeding industry, as the raw material of chromium nutrition element strengthening in feed; And other need the product strengthening chromium nutrition element.
Have the following advantages according to the product tool that the inventive method obtains:
● the chromium content of chromium content height Rich chromium yeast can reach more than 2000mg/kg, and its protein content reaches more than 45.0%, vitamins B
1reach 43.0mg/kg, vitamins B
2reaching 63.0mg/kg, is a kind of well functional food ingredient.
● the albumen of chromium in yeast in utilization ratio height Rich chromium yeast is organically combined, and its human absorptivity is far away higher than inorganic chromium.
● the toxicity of the low Rich chromium yeast of toxicity is significantly less than chromium trichloride.
Embodiment
Describe the embodiment of method of the present invention below in detail.
the amplification culture of bacterial classification
In the present invention, several times amplification culture can be carried out to yeast saccharomyces cerevisiae bacterial classification as required, to provide enough bacterial classifications.The number of times amplified depends on the scale that Rich chromium yeast ferments.In general, need bacterial classification to be carried out at least three times to amplify.Under the scale needed, 4 to 6 times can be carried out and amplify.
Normally, firsts and seconds amplifies is carry out in laboratory scale.Such as, in the Carlsberg's flask of the triangular flask of a 500ml and a 10L, one-level can be carried out respectively to amplify and secondary amplification.In the present invention, firsts and seconds amplifies the identical culture condition of employing, 28 ~ 31 DEG C and 4.8 ~ 5.8 pH value under cultivate, the substratum used there is no strict restriction, conventional carbon source (such as glucose, starch, sucrose and lactose), nitrogenous source (such as ammonium sulfate), and the additive of necessity such as VITAMIN, buffer reagent also can use.In one particular embodiment of the present invention, the substratum cultivated for one-level, secondary is made up of the aqueous sucrose solution of appropriate yeast powder, potassium primary phosphate and magnesium sulfate, wherein, the yeast powder of use can be the yeast extract of the type such as yeast extract, dried yeast powder.Typically, the substratum of use is the mixed aqueous solution containing 8 ~ 12% sucrose, 1 ~ 3% yeast powder, 0.1 ~ 0.5% potassium primary phosphate and 0.1 ~ 0.5% magnesium sulfate.In one embodiment of the invention, the substratum of use is the mixed aqueous solution containing 10% sucrose, 2% yeast powder, 0.1% potassium primary phosphate and 0.1% magnesium sulfate, and be 31 DEG C and 5.0 pH value under carry out.
With triangular flask by spawn culture suitable time (such as 40 ~ 48 hours) after, the nutrient solution obtained is transferred in the Carlsberg's flask that 7.5L substratum is housed, adopts same condition to carry out secondary cultivation.Secondary medium is originally upper is quiescent culture.Preferably, at second stage late stage of culture wave and culture container off and on, such as, within every 2 hours, Carlsberg's flask is shaken 2 minutes by shaking table.
Next, secondary is cultivated the bacterial classification obtained partly or entirely to transfer in a fermentor tank and carry out three grades of amplifications, to provide the bacterial classification of technical scale requirement.The time of volume needs needed for the scale of product fermentation (the last fermentation for the preparation of Rich chromium yeast) and three grades of amplifications of fermentor tank is designed, such as, for the bacterial classification of product fermentation by means of only three grades are amplified situation about providing, the bacterial classification that the volume of fermentor tank should be able to make third stage culture within the such as 20-48 hour suitable time obtain needed for product fermentation.
In the present invention, the substratum that these three grades are amplified is selected.Substratum needs to comprise carbon source, nitrogenous source, necessary mineral substance and optional other compositions, as VITAMIN and buffer reagent.Can be used for three grades of carbon sources of amplifying can be starch, glucose, sucrose and lactose etc.But consider from cost angle, preferably use treated sugar refinery byproduct---molasses.The molasses that the present invention uses can be the molasses of any kind, such as beet sirup or cane molasses.In an embodiment of the present invention, use cane molasses, but the operability of other molasses can not be got rid of.The molasses that buying comes before use generally will through dilution, heating, ventilate, leave standstill, be separated and the treatment step such as high-temperature sterilization, to be mixed with the production molasses containing about 30wt% fermentable sugar.In the present invention, elsewhere mentions that molasses are, typically refers to the molasses that sucrose content is 29%, unless otherwise specified.
The nitrogenous source that can be used in substratum of the present invention can be inorganic nitrogen-sourced and organic nitrogen source, wherein preferred organic nitrogen source, to accelerate biological growth.Inorganic nitrogen-sourcedly comprise ammonia, ammonium salt and nitrate etc., they should note when being employed fermenting in the change of pH, for three grades are amplified, generally pH value should be controlled 4.5 ~ 5.5, in the scope of active 4.8 ~ 5.2, most preferably 5.0.Organic nitrogen source comprises amino acid, protein, urea, soybean cake powder, groundnut meal, cottonseed meal, corn steep liquor, fish meal, yeast powder etc.Because considering cost factor, preferably use soybean cake powder, groundnut meal, cottonseed meal, corn steep liquor, fish meal, yeast powder as nitrogenous source.In a particular embodiment of the present invention, use yeast powder as nitrogenous source.Also organic nitrogen source and inorganic nitrogen-sourced combination can be used.Such as, yeast powder and primary ammonium phosphate can be used simultaneously, primary ammonium phosphate except as except buffer reagent, the double effect playing nitrogenous source.When using yeast powder, urea or ammonium sulfate or the combination of the two can also be added simultaneously.
In three grades of amplification culture, carbon source content is generally no more than 10wt%, and nitrogenous source content is lower, and general carbon-nitrogen ratio is 3 ~ 4: 1.Carbon and nitrogen sources content can not too high reason be avoid producing matrix or product to the suppression of reaction with cause cell dehydration because osmotic pressure is too high and dead.
Can add some other mineral substance in substratum of the present invention, as Mg, P, K, Ca and Zn etc. needed for supplementary biological growth, the example of these mineral substance includes but not limited to magnesium sulfate, zinc sulfate and phosphoric acid series matter, as hydrophosphate, phosphoric acid etc.In addition, the VITAMIN (such as VITMAIN B1) needed for seed growth and vitamin H can also be added.Because pH in fermenting process is very large on the impact forming biologic, for maintaining stable pH, often adopt buffer reagent, such as calcium carbonate or phosphoric acid salt.The latter's decapacitation regulates outside pH, also for substratum provides phosphorus source.
In an embodiment of the inventive method, comprise for three grades of substratum amplified: by the Weight computation of substratum, the molasses of 25% to 28%; The urea of 0.08% to 0.11%; The pH value of substratum is suitable for control the primary ammonium phosphate in 4.5 ~ 5.0 scopes; The yeast powder of 0.05% to 0.08%; The magnesium sulfate of 0.05% to 0.1%; And 0.02% to 0.05% zinc sulfate.
In the fermentation in this stage, need to fill into air in fermentor tank, to provide oxygen required in fermenting process.Usually, due to different in the yeast number of this grade of fermentation different time, therefore, air intake should work suitably changes according to the time of fermentation is different, such as, in the first half time of fermentation, can stronger ventilation speed gradually, and in the later half time, ventilation speed can remain unchanged.Also in whole fermenting process, ventilation speed can be amplified step by step.
In the present invention, this first step fermentation (three grades of amplifications), normally at 26 DEG C to 32 DEG C, is preferably carried out at the temperature of 29 DEG C to 31 DEG C.The time of fermentation can be decided according to biomass in the size of fermentor tank, leavening temperature and the unit volume fermented liquid set.In the present invention, it is suitable for carrying out when weight in wet base reaches 35-40g/L putting tank.Here, weight in wet base refers to the quality of often liter of fermented liquid by the fresh yeast of dry-matter near 26% of centrifugal rear acquisition.In one particular embodiment of the present invention, the design volume for three grades of fermentor tanks amplified is 10m
3, substratum is by 6m
3process water (water through sterilization), 2000L molasses (29% weight ratio), 9.1kg urea, 12.5kg primary ammonium phosphate, 4.8kg yeast powder, 4.8kg magnesium sulfate, 2.4kg zinc sulfate composition, for reaching 35 ~ 40g/L institute weight in wet base, need 31 DEG C of bottom fermentations 18 ~ 24 hours.For the fermentor tank of different size, can zoom in or out in this ratio.
product ferments
After completing three grades and amplifying, and after by second stage fermentor tank sterilizing, can proceed to disposable for fermented liquids all in three grade fermemtation tank in product fermentor tank.Also can only yeast-lactic isolated from fermented liquid be proceeded in product fermentor tank.But front a kind of mode is preferred, because in the present invention, the substratum difference that three grades of amplifications and product ferment used is little.The bacterial classification being in logarithmic phase can be made like this to be transplanted in control environment ferment, greatly shorten the lag phase of its process of growth.The reason that lag phase shortens ties up to seed growth phase formed owing to participating in the enzyme of metabolic activity in cells, and do not need spended time separately to build the enzyme system of suitable new environment.
According to method of the present invention, the substratum fermented for this product consists of the following components:
1) process water, namely through the water of sterilization;
2) based on the MgSO of process water weight 0.05% to 0.1%
4, the ZnSO of 0.05% to 0.1%
4, the KH of 0.08% to 0.12%
2pO
4;
3) molasses, its consumption is 75% for calculating standard with the transformation efficiency that molasses change into yeast.In a particular embodiment of the present invention, use vattability sugar content to be the cane molasses of 29wt%, but method of the present invention is not limited to cane molasses.
4) glucose mother liquid+molasses: it is by being that 30% ~ 40% glucose solution and molasses and appropriate water mix by homemade concentration, and adds 0.01 ~ 0.05%MgSO by weight
4with 0.01% ~ 0.05%ZnSO
4.Typically, in this part the ratio of each composition for being by glucose mother liquid (40%): molasses: water: MgSO
4: ZnSO
4=500L: 300L: 200L: 30kg: 30kg.
The molasses used in this step can be the molasses of any kind, such as beet sirup or cane molasses.In an embodiment of the present invention, use cane molasses, but the operability of other molasses can not be got rid of.
In a preferred embodiment of the present invention, product fermentation early stage, with 3) described in molasses as carbon source, in the later stage, with 4) in component replacement molasses as carbon source, be so more conducive to accumulation and the absorption of chromium.
5) N and P donor: may be used for N and P donor of the present invention and include but not limited to urea, NH
4h
2pO
4, (NH
4)
2sO
4, and yeast extract paste.In a preferred embodiment of the invention, for use 50m
3the situation of process water, employs 180kg urea, 126kg NH
4h
2pO
4, 300kg (NH
4)
2sO
4with 60kg yeast extract paste as N and P donor, be dissolved in 3000L water during use, disposablely before fermentation added in fermentor tank; And
6) trivalent chromium salt solution, it can be organo-chromium compound, such as chromium acetate, and also can be inorganic chromium compound, such as chromium trichloride, trivalent chromium compound used in the present invention includes but not limited to CrCl
36H
2o.In one particular embodiment of the present invention, for use 50m
3the situation of process water, employs 210kgCrCl
36H
2o, is dissolved in during use in 3000L water.
In addition, other nutritive ingredients appropriate can also be added in the medium, as vitamin H, calcium pantothenate, VITMAIN B1, Lin Suanna Vitamin B2 Sodium Phosphate and vitamin B6.In one particular embodiment of the present invention, for use 50m
3the situation of process water, before switching yeast starter, after adding 5 ~ 10g vitamin H, 300 ~ 450g calcium pantothenate, 150 ~ 200g VITMAIN B1,5 ~ 10g Lin Suanna Vitamin B2 Sodium Phosphate and 200 ~ 250g vitamin B6, then ferments to substratum.
It should be noted that, in the method for the invention, chromium cpd adds after product fermentation proceeds to a predefined phase, and doing like this is because chromium cpd is very large on yeast growth impact, if the fermentation initial stage adds will affect yeast growth, and causes final yeast output very low.Can ensure that again inorganic chromium compound is organic chromium by yeast conversion in a large number in order to reduce chromium cpd to the impact of yeast growth as far as possible, adding chromium cpd by the production advantageously in Rich chromium yeast in the fermentation middle and later periods.
In this operation, the content of chromium in fermented liquid should be controlled, its ultimate density is controlled between 800-1100ppm, otherwise will the growth of yeast cell be suppressed.In order to carry out product fermentation, three grades can be amplified all fermented liquids obtained that ferment transfers in the fermentor tank of size, then the process water, molasses and N and the P donor that prepare in advance is added, add the vitamin H of requirement, calcium pantothenate, VITMAIN B1, Lin Suanna Vitamin B2 Sodium Phosphate and vitamin B6 after sterilization, in suitable temperature as 31 DEG C of bottom fermentations.By detecting the weight in wet base of fermented liquid, after fermentation reaches a predetermined extent, appropriate halfcystine can be filled into, changing glucose and molasses mixed solution is carbon source simultaneously.After fermentation reaches another predetermined extent, by the pH regulator to 3.8 of fermented liquid ~ 4.0, be preferably adjusted to 3.9, then add with liquid stream the CrCl that form adds the above-mentioned preparation of requirement
3the aqueous solution.In adition process, remain this pH value range.
In the fermentation in this stage, need to fill into air in fermentor tank, to provide oxygen required in fermenting process.Usually, because the yeast number of the different time in fermentation is different, therefore, air intake should work suitably changes according to the time of fermentation is different, such as, in the first half time of fermentation, can stronger ventilation speed gradually, and in the later half time, ventilation speed can remain unchanged.Also in whole fermenting process, ventilation speed can be amplified step by step.
In the present invention, the fermentation of this product, normally at 28 DEG C to 37 DEG C, is preferably carried out at the temperature of 31 DEG C to 35 DEG C.PH value can be controlled between 3.8 to 6.8.The time of fermentation can be decided according to biomass in the size of fermentor tank, leavening temperature and the unit volume fermented liquid set.Those skilled in the art can regulate as the case may be.In one particular embodiment of the present invention, this product tank volume used of fermenting is designed to 90m
3, in this case, adopting above-mentioned fermentation system, at the fermentations of 31 DEG C DEG C, for reaching the weight in wet base of 150g/L, needing fermentation about 24 to 28 hours.
be separated
After completing the said products fermentation, adopt disk plate centrifuge to be separated fermented liquid, regulate by controlling fermented liquid feeding rate the yeast-lactic concentration being finally separated and reaching, the discharge of wastewater separated is to waste water workshop.The yeast-lactic obtained with technique water washing centrifugation, to obtain the whiter yeast-lactic of color.Being separated the yeast-lactic obtained adopts vacuum drum to filter, and then obtains the finished product by spraying dry, packaging and other steps.
The concrete enforcement of the inventive method is described with an embodiment below, but this embodiment only should be interpreted as and exemplifies of the present invention, and can not be used for limiting scope of the present invention.
Embodiment 1
The sucrose medium of 100 milliliters of sucrose containing 10% weight, 2% yeast powder, 0.1% potassium primary phosphate and 0.1% magnesium sulfate is introduced in the triangular flask of a 250ml, at PH 5.0, quiescent culture 48 hours at 31 DEG C.
In the Carlsberg's flask of one 15 liters, put into the sucrose medium 7.5L that fill a prescription same with described in the last period, after sterilizing, this Carlsberg's flask will be inoculated at the cultured yeast soln of triangular flask, PH 5.0,31 DEG C of quiescent culture 48 hours, later stage interval shake.
At a 10m
3the first fermentor tank in, use 6m
3process water, 2000L molasses (29%), 9.1kg urea, 12.5kg high purity phosphorus ammonium, 4.8kg yeast powder, 4.8kg magnesium sulfate, 2.4kg zinc sulfate are mixed with substratum, then the yeast soln that whole above-mentioned Carlsberg's flask is cultivated is seeded in this first fermentor tank, when weight in wet base reaches 35g--40g/L, stop fermentation, put tank.In the process, according to carrying out shown in table 1 fermenting and controlling to ventilate.
Table 1, the implementation condition of first step fermentation
| Time (h) | Temperature | pH | Stir | Ventilation M3/min |
| 0 | 31 | 5.0 | 0 | |
| 1 | 31 | 5.0 | 0 | |
| 2 | 31 | 5.0 | 0 | |
| 3 | 31 | 5.0 | 10 | |
| 4 | 31 | 5.0 | 10 | |
| 5 | 31 | 5.0 | 20 | |
| 6 | 31 | 5.0 | 40 | |
| 7 | 31 | 5.0 | 60 | |
| 8 | 31 | 5.0 | 80 | |
| 9 | 31 | 5.0 | 100 | |
| 10 | 31 | 5.0 | 100 | |
| 11 | 31 | 5.0 | 100 | |
| 12 | 31 | 5.0 | 100 | |
| 13 | 31 | 5.0 | 100 | |
| 14 | 31 | 5.0 | 100 | |
| 15 | 31 | 5.0 | 100 | |
| 16 | 31 | 5.0 | 100 | |
| 17 | 31 | 5.0 | 100 |
| 18 | 31 | 5.0 | 100 | |
| 19 | 31 | 5.0 | 100 | |
| 20 | 31 | 5.0 | 100 |
Following part solution is loaded a 90m
3the second fermentor tank in:
50m
3process water, and dissolve in 48kgMgSO
4, 48kgZnSO
4and 60kgKH
2pO
4;
Molasses: 7800L 29%
N and P donor: urea 180kg, NH
4h
2pO
4126kg, (NH
4)
2sO
4300kg, yeast extract paste 60kg is dissolved in 3000L water
After by fermentor tank sterilizing, add vitamin H 500g, calcium pantothenate 500g, VB
1500g, VB
2250g and VB
6250g.Then 9600L obtained above tri-grades being amplified fermented liquid transfers in this fermentor tank, and the condition listed according to table 2 is carried out fermenting and controlled to ventilate.
Table 2, the implementation condition of second stage fermentation
| H | Molasses (L/hr) | Nitrogen .P (L/hr) | Chromium source L | Temperature | PH | Ventilate (m3/min) | Alcohol | Remarks |
| 0 | 0 | 0 | 31 | 4.2 | 2000 | 0.60 | ||
| 1 | 75 | 60 | 31 | 4.2 | 3000 | 0.60 | ||
| 2 | 118 | 60 | 31 | 4.5 | 3000 | 0.60 | ||
| 3 | 187 | 120 | 31 | 4.5 | 3000 | 0.60 | ||
| 4 | 220 | 120 | 31 | 4.8 | 4000 | 0.60 | ||
| 5 | 279 | 120 | 31 | 4.8 | 4000 | 0.60 | ||
| 6 | 279 | 120 | 31 | 4.8 | 4000 | 0.60 | ||
| 7 | 325 | 120 | 31 | 4.8 | 5000 | 0.60 | ||
| 8 | 398 | 180 | 31 | 4.8 | 5000 | 0.60 | ||
| 9 | 488 | 180 | 31 | 4.8 | 6000 | 0.60 | ||
| 10 | 524 | 180 | 31 | 4.5 | 6000 | 0.60 | ||
| 11 | 587 | 180 | 31 | 4.2 | 7000 | 0.60 | ||
| 12 | 700 | 240 | 480 | 31 | 3.9 | 8000 | 0.60 | |
| 13 | 840 | 240 | 480 | 31 | 3.9 | 8000 | 0.60 | |
| 14 | 1080 | 240 | 480 | 31 | 3.9 | 10000 | 0.60 | |
| 15 | 1080 | 240 | 480 | 31 | 3.9 | 10000 | 0.60 | |
| 16 | 1320 | 240 | 480 | 31 | 3.9 | 10000 | 0.60 |
| 17 | 1500 | 180 | 600 | 31 | 3.9 | 10000 | 0.60 | |
| 18 | 1500 | 180 | 31 | 3.9 | 10000 | 0.60 | ||
| 19 | 1500 | 31 | 3.9 | 10000 | 0.60 | |||
| 20 | 1800 | 31 | 3.9 | 10000 | 0.50 | |||
| 21 | 1440 | 31 | 3.9 | 10000 | ＜0.8 | |||
| 22 | 1440 | 31 | 3.9 | 10000 | ＜0.8 | |||
| 23 | 1000 | 31 | 3.9 | 10000 | ＜0.8 | |||
| 23 | 720 | 31 | 3.9 | 10000 | ＜0.8 | |||
| 24 | 31 | 3.9 | 10000 |
In the fermentation of this product, add halfcystine 36kg when weight in wet base is 50g/L, once finish, change glucose and molasses mixed solution is carbon source simultaneously.The concrete compound method of this carbon source is, 40wt% glucose solution: molasses: water: MgSO
4: ZnSO
4=500L: 300L: 200L: 30kg: 30kg.Configuration 12000L.
Start when weight in wet base is 65g/L to add chromium source, it is by by 210kg CrCl
36H
2o is dissolved in 3000L water, adds with liquid stream form.Adding fashionable maintenance PH is 3.9, adds in 6 hours by technological design.When weight in wet base reaches 120g/L, stop fermentation, put tank.
After putting tank, rinse fermentor tank with 0.1%EDTA, the chromium complexing be attached in fermentor tank is got off.Adopt disk plate centrifuge to be separated fermented liquid, the yeast-lactic process water obtained carries out washing and is separated, to obtain the whiter yeast-lactic of color.Being separated the yeast-lactic obtained adopts vacuum drum to filter, and then by spraying dry, obtains about ... the yellow the finished product of .kg, wherein dry-matter 95.34%, chromium content is 2053ppm, is 2153ppm after giving money as a gift, its protein content reaches more than 45.0%, vitamins B
1reach 43.0mg/kg, vitamins B
2reach 63.0mg/kg.
Embodiment 2
The sucrose medium of 100 milliliters of sucrose containing 8% weight, 1.5% yeast powder, 0.1% potassium primary phosphate and 0.1% magnesium sulfate is introduced in the triangular flask of a 250ml, at PH4.9, quiescent culture 48 hours at 29 DEG C.
In the Carlsberg's flask of one 15 liters, put into the sucrose medium 7.5L that fill a prescription same with described in the last period, after sterilizing, this Carlsberg's flask will be inoculated at the cultured yeast soln of triangular flask, with the CMC model identical with previous step 48 hours, later stage interval shake.
At a 10m
3the first fermentor tank in, use 6m
3process water, 2500L molasses (containing sucrose amount 29%), 6.8kg urea, 6.2kg high purity phosphorus ammonium, 4.4kg yeast powder, 4.4kg magnesium sulfate, 2.2kg zinc sulfate are mixed with substratum, then the yeast soln that whole above-mentioned Carlsberg's flask is cultivated is seeded in this first fermentor tank, when weight in wet base reaches 35g--40g/L, stop fermentation, put tank.In the process, according to carrying out shown in table 3 fermenting and controlling to ventilate.
Table 3, the implementation condition of first step fermentation
| Time (h) | Temperature | pH | Stir | Ventilation M3/min |
| 0 | 28 | 4.6 | 0 | |
| 1 | 28 | 4.6 | 0 | |
| 2 | 28 | 4.6 | 0 | |
| 3 | 28 | 4.6 | 10 | |
| 4 | 28 | 4.6 | 10 | |
| 5 | 28 | 4.6 | 20 | |
| 6 | 28 | 4.6 | 40 | |
| 7 | 28 | 4.6 | 60 | |
| 8 | 28 | 4.6 | 80 | |
| 9 | 28 | 4.6 | 100 | |
| 10 | 28 | 4.6 | 100 | |
| 11 | 28 | 4.6 | 100 | |
| 12 | 28 | 4.6 | 100 | |
| 13 | 28 | 4.6 | 100 | |
| 14 | 28 | 4.6 | 100 | |
| 15 | 28 | 4.6 | 100 | |
| 16 | 28 | 4.6 | 100 | |
| 17 | 28 | 4.6 | 100 | |
| 18 | 28 | 4.6 | 100 | |
| 19 | 28 | 4.6 | 100 | |
| 28 | 28 | 4.6 | 100 |
Following part solution is loaded a 90m
3the second fermentor tank in:
50m
3process water, and dissolve in 25kgMgSO
4, 25kgZnSO
4and 40kgKH
2pO
4;
Molasses: 7800L 29%
N and P donor: urea 130kg, NH
4h
2pO
4126kg, (NH
4)
2sO
4270kg, yeast extract paste 60kg is dissolved in 3000L water.
After by fermentor tank sterilizing, add vitamin H 500g, calcium pantothenate 500g, VB
1500g, VB
2250g and VB
6250g.Then 9600L obtained above tri-grades being amplified fermented liquid transfers in this fermentor tank, and the condition listed according to table 4 is carried out fermenting and controlled to ventilate.
Table 4, the implementation condition of second stage fermentation
| H | Molasses (L/hr) | Nitrogen .P (L/hr) | Chromium source L | Temperature | PH | Ventilate (m 3/min) | Alcohol | Remarks |
| 0 | 0 | 0 | 28～31 | 6.2 | 2000 | 0.60 | ||
| 1 | 75 | 60 | 28～31 | 6.2 | 3000 | 0.60 | ||
| 2 | 118 | 60 | 28～31 | 6.5 | 3000 | 0.60 | ||
| 3 | 187 | 120 | 28～31 | 6.5 | 3000 | 0.60 | ||
| 4 | 220 | 120 | 28～31 | 6.8 | 4000 | 0.60 | ||
| 5 | 279 | 120 | 28～31 | 6.8 | 4000 | 0.60 | ||
| 6 | 279 | 120 | 28～31 | 6.8 | 4000 | 0.60 | ||
| 7 | 325 | 120 | 28～31 | 6.8 | 5000 | 0.60 | ||
| 8 | 398 | 180 | 28～31 | 6.8 | 5000 | 0.60 | ||
| 9 | 488 | 180 | 28～31 | 6.8 | 6000 | 0.60 | ||
| 10 | 524 | 180 | 28～31 | 6.5 | 6000 | 0.60 | ||
| 11 | 587 | 180 | 28～31 | 6.2 | 7000 | 0.60 | ||
| 12 | 700 | 240 | 480 | 28～31 | 5.9 | 8000 | 0.60 | |
| 13 | 840 | 240 | 480 | 28～31 | 5.9 | 8000 | 0.60 | |
| 14 | 1080 | 240 | 480 | 28～31 | 5.9 | 10000 | 0.60 | |
| 15 | 1080 | 240 | 480 | 28～31 | 5.9 | 10000 | 0.60 | |
| 16 | 1320 | 240 | 480 | 28～31 | 5.9 | 10000 | 0.60 | |
| 17 | 1500 | 180 | 600 | 28～31 | 5.9 | 10000 | 0.60 | |
| 18 | 1500 | 180 | 28～31 | 5.9 | 10000 | 0.60 | ||
| 19 | 1500 | 28～31 | 5.9 | 10000 | 0.60 |
| 20 | 1800 | 28～31 | 5.9 | 10000 | 0.50 | |||
| 21 | 1440 | 28～31 | 5.9 | 10000 | ＜0.8 | |||
| 22 | 1440 | 28～31 | 5.9 | 10000 | ＜0.8 | |||
| 23 | 1000 | 28～31 | 5.9 | 10000 | ＜0.8 | |||
| 23 | 720 | 28～31 | 5.9 | 10000 | ＜0.8 | |||
| 24 | 28～31 | 5.9 | 10000 |
In the fermentation of this product, add halfcystine 36kg when weight in wet base is 50g/L, once finish, change glucose and molasses mixed solution is carbon source simultaneously.The concrete compound method of this carbon source is, 40wt% glucose solution: molasses: water: MgSO
4: ZnSO
4=500L: 300L: 200L: 30kg: 30kg, proportions 12000L like this.
Start when weight in wet base is 35g/L to add chromium source, it is by by 41.5kgCrCl
36H
2o is dissolved in 3000L water, adds with liquid stream form.Adding fashionable maintenance PH is 5.9, adds in 6 hours by technological design.When weight in wet base reaches 110g/L, stop fermentation, put tank.
After putting tank, rinse fermentor tank with 0.1%EDTA, the chromium complexing be attached in fermentor tank is got off.Adopt disk plate centrifuge to be separated fermented liquid, the yeast-lactic process water obtained carries out washing and is separated, to obtain the whiter yeast-lactic of color.Being separated the yeast-lactic obtained adopts vacuum drum to filter, and then by spraying dry, obtain yellow the finished product, wherein dry-matter 92.15%, chromium content is 2002ppm, and be 2103ppm after giving money as a gift, its protein content reaches more than 45.0%, vitamins B
1reach 43.0mg/kg, vitamins B
2reach 63.0mg/kg.
Embodiment 3
The sucrose medium of 100 milliliters of sucrose containing 11% weight, 2.5% yeast powder, 0.5% potassium primary phosphate and 0.5% magnesium sulfate is introduced in the triangular flask of a 250ml, at PH5.5, quiescent culture 36 hours at 31 DEG C.
In the Carlsberg's flask of one 15 liters, put into the sucrose medium 7.5L that fill a prescription same with described in the last period, after sterilizing, this Carlsberg's flask will be inoculated at the cultured yeast soln of triangular flask, with the CMC model identical with previous step 48 hours, later stage interval shake.
At a 10m
3the first fermentor tank in, use 6m
3process water, 2200L molasses (containing sucrose amount 29%), 8.8kg urea, 12.8kg high purity phosphorus ammonium, 6.4kg yeast powder, 8.8kg magnesium sulfate, 4.2kg zinc sulfate are mixed with substratum, then the yeast soln that whole above-mentioned Carlsberg's flask is cultivated is seeded in this first fermentor tank, when weight in wet base reaches 35g--40g/L, stop fermentation, put tank.In the process, according to carrying out shown in table 5 fermenting and controlling to ventilate.
Table 5, the implementation condition of first step fermentation
| Time (h) | Temperature | pH | Stir | Ventilation M3/min |
| 0 | 32 | 5.4 | 0 | |
| 1 | 32 | 5.4 | 0 | |
| 2 | 32 | 5.4 | 0 | |
| 3 | 32 | 5.4 | 10 | |
| 4 | 32 | 5.4 | 10 | |
| 5 | 32 | 5.4 | 20 | |
| 6 | 32 | 5.4 | 40 | |
| 7 | 32 | 5.4 | 60 | |
| 8 | 32 | 5.4 | 80 | |
| 9 | 32 | 5.4 | 100 | |
| 10 | 32 | 5.4 | 100 | |
| 11 | 32 | 5.4 | 100 | |
| 12 | 32 | 5.4 | 100 | |
| 13 | 32 | 5.4 | 100 | |
| 14 | 32 | 5.4 | 100 | |
| 15 | 32 | 5.4 | 100 | |
| 16 | 32 | 5.4 | 100 | |
| 17 | 32 | 5.4 | 100 | |
| 18 | 32 | 5.4 | 100 | |
| 19 | 32 | 5.4 | 100 | |
| 28 | 32 | 5.4 | 100 |
Following part solution is loaded a 90m
3the second fermentor tank in:
50m
3process water, and dissolve in 50kgMgSO
4, 50kgZnSO
4and 60kgKH
2pO
4;
Molasses: 7800L 29%
N and P donor: urea 240kg, NH
4h
2pO
4240kg, (NH
4)
2sO
4360kg, yeast extract paste 120kg is dissolved in 3000L water
After by fermentor tank sterilizing, add vitamin H 500g, calcium pantothenate 500g, VB
1500g, VB
2250g and VB
6250g.Then 9600L obtained above tri-grades being amplified fermented liquid transfers in this fermentor tank, and the condition listed according to table 6 is carried out fermenting and controlled to ventilate.
Table 6, the implementation condition of second stage fermentation
| H | Molasses (L/hr) | Nitrogen .P (L/hr) | Chromium source L | Temperature | PH | Ventilate (m3/min) | Alcohol | Remarks |
| 0 | 0 | 0 | 31～34 | 4.2 | 2000 | 0.60 | ||
| 1 | 75 | 60 | 31～34 | 4.2 | 3000 | 0.60 | ||
| 2 | 118 | 60 | 31～34 | 4.5 | 3000 | 0.60 | ||
| 3 | 187 | 120 | 31～34 | 4.5 | 3000 | 0.60 | ||
| 4 | 220 | 120 | 31～34 | 4.8 | 4000 | 0.60 | ||
| 5 | 279 | 120 | 31～34 | 4.8 | 4000 | 0.60 | ||
| 6 | 279 | 120 | 31～34 | 4.8 | 4000 | 0.60 | ||
| 7 | 325 | 120 | 31～34 | 4.8 | 5000 | 0.60 | ||
| 8 | 398 | 180 | 31～34 | 4.8 | 5000 | 0.60 | ||
| 9 | 488 | 180 | 31～34 | 4.8 | 6000 | 0.60 | ||
| 10 | 524 | 180 | 31～34 | 4.5 | 6000 | 0.60 | ||
| 11 | 587 | 180 | 31～34 | 4.2 | 7000 | 0.60 | ||
| 12 | 700 | 240 | 480 | 31～34 | 3.9 | 8000 | 0.60 | |
| 13 | 840 | 240 | 480 | 31～34 | 3.9 | 8000 | 0.60 | |
| 14 | 1080 | 240 | 480 | 31～34 | 3.9 | 10000 | 0.60 | |
| 15 | 1080 | 240 | 480 | 31～34 | 3.9 | 10000 | 0.60 | |
| 16 | 1320 | 240 | 480 | 31～34 | 3.9 | 10000 | 0.60 | |
| 17 | 1500 | 180 | 600 | 31～34 | 3.9 | 10000 | 0.60 | |
| 18 | 1500 | 180 | 31～34 | 3.9 | 10000 | 0.60 | ||
| 19 | 1500 | 31～34 | 3.9 | 10000 | 0.60 |
| 20 | 1800 | 31～34 | 3.9 | 10000 | 0.50 | |||
| 21 | 1440 | 31～34 | 3.9 | 10000 | ＜0.8 | |||
| 22 | 1440 | 31～34 | 3.9 | 10000 | ＜0.8 | |||
| 23 | 1000 | 31～34 | 3.9 | 10000 | ＜0.8 | |||
| 23 | 720 | 31～34 | 3.9 | 10000 | ＜0.8 | |||
| 24 | 31～34 | 3.9 | 10000 |
In the fermentation of this product, add halfcystine 36kg when weight in wet base is 50g/L, once finish, change glucose and molasses mixed solution is carbon source simultaneously.The concrete compound method of this carbon source is, 40wt% glucose solution: molasses: water: MgSO
4: ZnSO
4=500L: 300L: 200L: 30kg: 30kg, preparation 12000L.
Start when weight in wet base is 65g/L to add chromium source, it is by by 415kg CrCl
36H
2o is dissolved in 3000L water, adds with liquid stream form.Adding fashionable maintenance PH is 3.9, adds in 6 hours by technological design.When weight in wet base reaches 150g/L, stop fermentation, put tank.
After putting tank, rinse fermentor tank with 0.1%EDTA, the chromium complexing be attached in fermentor tank is got off.Adopt disk plate centrifuge to be separated fermented liquid, the yeast-lactic process water obtained carries out washing and is separated, to obtain the whiter yeast-lactic of color.Being separated the yeast-lactic obtained adopts vacuum drum to filter, and then by spraying dry, obtain yellow the finished product, wherein dry-matter 94.34%, chromium content is 2075ppm, and be 2174ppm after giving money as a gift, its protein content reaches more than 45.0%, vitamins B
1reach 43.0mg/kg, vitamins B
2reach 63.0mg/kg.
Because the present invention selects add chromic salts a certain suitable opportunity in saccharomycetes to make fermentation process, both the growth suppressing yeast because of adding of chromic salts had been avoided, also be conducive to the absorption of yeast to chromic salts, thus produce high chromium content, safety non-toxic and be beneficial to animal body absorb Rich chromium yeast product.
According to the standard of " protective foods inspection and assessment technical specifications " (2003 editions), carry out toxicity inspection (Inspection Unit: function of health food inspection center of Tongji Medical College, Huazhong Science and Technology Univ.) to the product of embodiment 1 gained, the conclusion drawn is as follows: this Rich chromium yeast recommended intake is 3.33mg/kg.bw.The acute toxicity tests shows, the mld (LD50) of this Rich chromium yeast to mouse is greater than 10.0g/kg.kw, belongs to actual non-toxic substance.The test of Salmonella reversion test, Micronuclei In The Mouse Bone Marrow is feminine gender with sperm malformation test.In rat 30 days feeding studys, animal is given by 25 times, 50 times, 100 times (are respectively 83,167,333mg/kg.kw) of human body recommended intake, and set up Normal group and wine base control group, measurement result shows that the indexs such as each group of rat body weight growth, food utilization, dirty body ratio, hematology and serum biochemistry are all in normal range, and each dosage group does not have significance with the corresponding group difference that contrasts; The poisoning pathology relevant with tested material is not found in the inspection of liver,kidney,spleen, stomach and intestine, ovary and testopathy yet.According to " protective foods inspection and assessment technical specifications " (2003 editions), judge that this Angel TH Rich chromium yeast is checked by food safety toxicology.
As can be seen here, adopt the Rich chromium yeast that method of the present invention is produced, achieve the conversion of inorganic chromium to organic chromium well, improve the specific absorption of chromium in animal body, thus be that human body and animal benefit chromium provide approach.
The raw material mending chromium protective foods and medicine is can be used as according to the Rich chromium yeast that the inventive method obtains; Can be used as the raw material of chromium nutrition element strengthening in nourishing reinforced food; Can be used for livestock breeding industry, as the raw material of chromium nutrition element strengthening in feed; And other need the product strengthening chromium nutrition element.
Have the following advantages according to the product tool that the inventive method obtains:
● the chromium content of chromium content height Rich chromium yeast can reach more than 2000mg/kg, and its protein content reaches more than 45.0%, vitamins B
1reach 43.0mg/kg, vitamins B
2reaching 63.0mg/kg, is a kind of well functional food ingredient.
● the albumen of chromium in yeast in utilization ratio height Rich chromium yeast is organically combined, and its human absorptivity is far away higher than inorganic chromium.
● the toxicity of the low Rich chromium yeast of toxicity is significantly less than chromium trichloride.
Above to the description of specific embodiment of the present invention and embodiment be for illustration of with the object described, although described the present invention by some previous embodiment, should not regard limitation of the present invention as.They are not detailed or limit the invention to disclosed precise forms, and obviously according to above-mentioned instruction, many improvement, embodiment and change are possible.Therefore, scope of the present invention comprises general field as herein disclosed, and contained by claims and its equivalence replacement.
Claims (17)
1. produce a method for Rich chromium yeast, comprise the following steps:
By yeast saccharomyces cerevisiae bacterial classification amplification culture several times, to provide the yeast saccharomyces cerevisiae bacterial classification of aequum;
Being incorporated in a fermentor tank by the yeast saccharomyces cerevisiae bacterial classification of described aequum, is 3.8 ~ 6.8 at pH, and temperature is the condition bottom fermentation of 28 ~ 35 DEG C, and when weight in wet base reaches more than 30g/L, stream adds chromium salt solution, ferments when reaching more than 110g/L to weight in wet base and stops;
By centrifugation, washing and dry results Rich chromium yeast product,
Wherein, described chromic salts is CrCl
36H
2o, its consumption is, every 100ml fermented liquid uses 0.05 ~ 0.5g chromic salts.
2. method according to claim 1, wherein, using the substratum containing following ingredients as initial medium:
1) process water;
2) MgSO
4, ZnSO
4, and KH
2pO
4;
3) molasses;
4) N and P donor solution, it contains urea, NH
4h
2p0
4, (NH
4)
2sO
4, and yeast extract paste.
3. method according to claim 2, wherein, based on described process water Weight computation, described MgSO
4consumption is 0.05% to 0.1%, described ZnSO
4consumption is 0.05% to 0.1%, and described KH
2pO
4consumption be 0.08% to 0.12%.
4. method according to claim 2, wherein, described N and P donor is the solution containing following ingredients: process water, and based on the % urea of 0.4 ~ 0.8, the NH of 0.4 ~ 0.8% of process water weight
4h
2p0
4, 0.8 ~ 1.2% (NH
4)
2sO
4, and 0.2 ~ 0.4% yeast extract paste.
5. method according to claim 2, wherein, described initial medium is further containing vitamin H, calcium pantothenate, VITMAIN B1, Lin Suanna Vitamin B2 Sodium Phosphate and vitamin B6.
6. method according to any one of claim 1 to 5, wherein, adds halfcystine when described weight in wet base reaches 40-70g/L, and with by water, glucose mother liquid, described molasses, MgSO
4with, ZnSO
4the solution carbon source as a supplement of composition.
7. method according to claim 6, wherein, the configuration proportion of the described solution of carbon source is as a supplement: 40wt% Pu grape Tang Rong Ye ︰ Tang Mi ︰ Shui ︰ MgSO
4︰ ZnSO
4=500L ︰ 300L ︰ 200L ︰ 30kg ︰ 30kg.
8. method according to claim 1, wherein, by described yeast saccharomyces cerevisiae bacterial classification through three amplification culture, wherein, one-level amplify and secondary amplification be 28 ~ 31 DEG C and 4.8 ~ 5.8 pH value under carry out.
9. method according to claim 8 wherein, is the mixed aqueous solution containing 8 ~ 12% sucrose, 1 ~ 3% yeast powder, 0.1 ~ 0.5% potassium primary phosphate and 0.1 ~ 0.5% magnesium sulfate for the substratum that described one-level is amplified and secondary amplifies.
10. method according to claim 8, wherein, three grades of amplifications carry out under being the condition of 4.5 to 5.5, until stop when weight in wet base reaches 35-40g/L at temperature, the pH of 26 to 32 DEG C.
11. methods according to claim 10, wherein, pressing the Weight computation of substratum for described three grades of substratum amplified, is by the molasses of 25% to 28%; The urea of 0.08% to 0.11%; The primary ammonium phosphate of 0.07% to 0.16%; The yeast powder of 0.05% to 0.08%; The magnesium sulfate of 0.05% to 0.1%; The process water composition of 0.02% to 0.05% zinc sulfate and surplus.
12. methods according to claim 1, wherein, ferment stops when weight in wet base reaches 150g/L.
13. methods according to claim 6, wherein, described solution contains the glucose of 15 ~ 25%, the vattability sugar content of 25 ~ 35% is 29% molasses, 0.01 ~ 0.05%MgSO
4with 0.01% ~ 0.05%ZnSO
4.
14. 1 kinds of methods of producing Rich chromium yeast, comprise the following steps:
A) in a triangular flask, and 28 ~ 31 DEG C and 4.8 ~ 5.8 pH value under yeast saccharomyces cerevisiae bacterial classification carried out 36 ~ 48 hours one-levels and amplifies, the substratum amplified for described one-level is the mixed aqueous solution containing 8 ~ 12% sucrose, 1 ~ 3% yeast powder, 0.1 ~ 0.5% potassium primary phosphate and 0.1 ~ 0.5% magnesium sulfate;
B) gained fermented liquid is proceeded in a Carlsberg's flask carry out 36 ~ 48 hours secondarys and amplify, its 28 ~ 31 DEG C and 4.8 ~ 5.8 pH value condition under carry out, the substratum amplified for described secondary is the mixed aqueous solution containing 8 ~ 12% sucrose, 1 ~ 3% yeast powder, 0.1 ~ 0.5% potassium primary phosphate and 0.1 ~ 0.5% magnesium sulfate;
C) gained fermented liquid is proceeded to the first fermentor tank and carry out three grades of amplifications, described three grades of amplifications carry out under being the condition of 4.5 to 5.5 at temperature, the pH of 26 to 32 DEG C, until stop when weight in wet base reaches 35-40g/L, wherein, pressing the Weight computation of substratum for described three grades of substratum amplified, is by the molasses of 25% to 28%; The urea of 0.08% to 0.11%; The primary ammonium phosphate of 0.07% to 0.16%; The yeast powder of 0.05% to 0.08%; The magnesium sulfate of 0.05% to 0.1%; The process water composition of 0.02% to 0.05% zinc sulfate and surplus;
D) gained fermented liquid is proceeded to one second fermentor tank and carry out product fermentation, wherein said product fermentation is at 28 DEG C to 35 DEG C, and pH carries out under the condition of 3.8 to 6.8, and the initial medium fermented for described product comprises:
Process water, and based on the MgSO of process water weight 0.05% to 0.1%
4, the ZnSO of 0.05% to 0.1%
4, and the KH of 0.08% to 0.12%
2pO
4;
Molasses;
N and P donor solution: it is the solution containing following ingredients: process water, and based on the % urea of 4 ~ 8, the NH of 4 ~ 8% of process water weight
4h
2p0
4, 8 ~ 12% (NH
4)
2sO
4with 2 ~ 4% yeast extract paste;
Appropriate vitamin H, calcium pantothenate, VITMAIN B1, Lin Suanna Vitamin B2 Sodium Phosphate and vitamin B6,
Wherein, the volume ratio of described process water, molasses and described N and P donor solution is 50 ︰ 12 ︰ 3,
Wherein, add appropriate halfcystine when described weight in wet base reaches 50g/L, and with by water, glucose mother liquid, described molasses, MgSO
4with, ZnSO
4the solution carbon source as a supplement of composition, and when described weight in wet base is more than 30g/L, stream adds CrCl
36H
2o solution, wherein said CrCl
36H
2the consumption of O is that every 100ml fermented liquid uses 0.05 ~ 0.5g, ferments to stop when weight in wet base reaches 110 ~ 150g/L; And
E) separation, washing, dry gained Rich chromium yeast product.
15. methods according to claim 14, comprise the following steps:
A) in a triangular flask, and 29 DEG C and 4.9 pH value under yeast saccharomyces cerevisiae bacterial classification carried out 48 hours one-levels and amplifies, the substratum amplified for described one-level is the mixed aqueous solution containing 8% sucrose, 1.5% yeast powder, 0.1% potassium primary phosphate and 0.1% magnesium sulfate;
B) gained fermented liquid is proceeded in a Carlsberg's flask carry out 48 hours secondarys and amplify, its 29 DEG C and 4.9 pH value under carry out, the substratum amplified for described secondary is the mixed aqueous solution containing 8% sucrose, 1.5% yeast powder, 0.1% potassium primary phosphate and 0.1% magnesium sulfate;
C) gained fermented liquid is proceeded to the first fermentor tank and carry out three grades of amplifications, described three grades of amplifications carry out under being the condition of 4.9 at temperature, the pH of 29 DEG C, until stop when weight in wet base reaches 35-40g/L, wherein, is: every 6m for described three grades of substratum amplified
3process water uses 2500L vattability sugar content to be the molasses of 29%; 6.8kg urea; 6.2kg primary ammonium phosphate; 4.4kg yeast powder; 4.4kg magnesium sulfate; 2.2kg zinc sulfate;
D) gained fermented liquid is proceeded to the second fermentor tank and carry out product fermentation, wherein said product fermentation is at 28 DEG C, and pH carries out under the condition of 3.8 to 4.9, and according to such proportions initial medium, every 9600L is amplified to the fermented liquid of gained by above-mentioned three grades, introduces:
Process water: 50m
3,
MgSO
4，25kg，
ZnSO
4：25kg，
KH
2PO
4：40kg
Wherein, vattability sugar content is 29% to molasses: 7800L,
N and P donor solution: it is by 130kg urea, 126kg NH
4h
2p0
4, 270kg (NH
4)
2sO
4, and 60kg yeast extract paste is dissolved in 3000L water and makes,
Vitamin H: 500g
Calcium pantothenate: 500g
VB1：500g
VB2:250g, and
VB6：250g
Wherein, add appropriate halfcystine when described weight in wet base reaches 50g/L, and with following solution carbon source as a supplement, the specific configuration method of this supplementary carbon source is: the sweet ︰ water ︰ MgSO of the molten liquid ︰ sugar of 40wt% Portugal grape sugar
4︰ ZnSO
4=500L ︰ 300L ︰ 200L ︰ 30kg ︰ 30kg, proportional arrangement 12000L like this,
Further, when described weight in wet base is more than 35g/L, stream adds chromium salt solution, and pH is adjusted to 5.9, and wherein, described chromium salt solution is by 41.5kgCrCl
36H
2o and 3000L process water forms, and ferment stops when weight in wet base reaches 110g/L; And
E) separation, washing, dry gained Rich chromium yeast product.
16. methods according to claim 14, wherein, steps d) in described product fermentation be at 28 DEG C to 35 DEG C, pH carries out under the condition of 3.8 to 6.8.
17. methods according to claim 14, wherein, as a supplement the described solution of carbon source contain 15 ~ 25% glucose, 25 ~ 35% vattability sugar content be 29% molasses, 0.01 ~ 0.05%MgSO
4with 0.01% ~ 0.05%ZnSO
4.
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