CN101575580B - Zinc-rich saccharomyces cerevisiae and production method thereof - Google Patents
Zinc-rich saccharomyces cerevisiae and production method thereof Download PDFInfo
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Abstract
The invention provides a zinc-rich saccharomyces cerevisiae Z1.4CCTCC M 205126. The zinc-rich saccharomyces cerevisiae has stable zinc resistance, fast growing speed and high utility ratio of zinc andcontains zinc as high as 80,000mg/kg, wherein the content of biological zinc can reach 90 percent.
Description
Technical field
The present invention relates to a kind of mikrobe, relate in particular to a kind of Zinc-rich saccharomyces cerevisiae, the invention still further relates to the selection of this bacterial classification and use this bacterial classification to carry out industrial method.
Background technology
Zinc is the essential trace element of human body and animal, and in vivo, zinc is the moity of many enzymes, can influence the mechanism configuration of the organic molecule coordination body of some non-enzyme again, plays important effect to keeping the normal physiological function of human body.Human body lacks zinc can cause numerous disease, like nanism, mellitus, hypertension, sexual organ and secondal sexual character underdevelopment, diseases such as male sterility.A part of zinc is lost in the normal physiological metabolism of human body meeting, and the zinc that replenishes in the food is not enough to satisfy the zinc of the homergy of human body, thereby causes people generally to lack zinc.Zinc-rich saccharomyces cerevisiae is as a kind of good zinc supplementation raw material, and its bioavailability is high, and toxic side effect is little, and contains rich in amino acid and VITAMINs in the yeast, and these can both promote the absorption of zinc.
Many to Zinc-rich saccharomyces cerevisiae research both at home and abroad in recent years; But content is not very high; Main problem is: (1) is not enough to the bacterial screening of Zinc-rich saccharomyces cerevisiae, and the bacterial classification that screens is not strong to the accumulation ability of zinc, and the zinc content in the yeast not high (2) organic zinc content is low; Part just yeast and simple absorption of zinc or mixing are arranged, inorganic zinc is not converted into organic zinc; (3) certain methods is not suitable for big production, and repeatability is very poor, and some can be produced, but productive rate is very low, and production cost is higher; (4) utilization ratio to zinc is low.
Summary of the invention
The objective of the invention is to provides the higher Zinc-rich saccharomyces cerevisiae bacterial classification of a kind of zinc content and utilizes this bacterial classification to carry out industrial method to above-mentioned deficiency.
Because having certain inhibition, general yeast to be difficult under the condition that contains high zinc to zymic growth, inorganic zinc grows, even growth velocity is arranged but not high to the utilization ratio of zinc.The present invention is through screening, domestication, mutagenesis, selects that zinc content is high, growth velocity is fast, productive rate is high, the Zinc-rich saccharomyces cerevisiae bacterial classification high to the utilization ratio of zinc, wherein a preferred strain called after Z1.4.This yeast strain on October 25th, 2005 at China typical culture collection center (CCTCC; Wuhan University; Postcode 430072) carried out biological preservation, classification called after yeast saccharomyces cerevisiae (Saccharomyces cerevisiae), deposit number is CCTCC M205126.
Specifically, through following method breeding species:
1) barms of the anti-high zinc of screening: different barmses is inoculated into containing in the zinc substratum of different concns, the bacterial classification that screening can be grown in high zinc substratum.Pick out the bacterial classification of the bacterial classification of anti-zinc as domestication.
2) bacterial classification inoculation of anti-zinc is cultivated to the substratum of high density zinc, and improved zinc concentration in the substratum gradually, keep zinc in higher concentration.Pick out anti-high zinc and stable bacterial classification as the mutagenesis bacterial classification.
3) with the mutagenesis in the substratum of the lithium chloride of different concns of select bacterial classification inoculation.
4) bacterial classification with mutagenesis carries out separation screening again, utilize shake bottle ferment with the small-sized fermentation jar select zinc content height, growth velocity is fast, productive rate is high, to the barms of the high anti-high zinc of the utilization ratio of zinc, be the Zinc-rich saccharomyces cerevisiae bacterial classification.
5) screening and culturing based formulas
Malt extract medium sugar degree 5～25%, containing zinc (zinc-amino acid chelate) concentration is 150～5000 μ g/ml, the pH value is 4～6.5.
6) screening and culturing condition
26～35 ℃ of temperature, stir culture 18～48 hours.
Another aspect the invention provides the method for producing above-mentioned Zinc-rich saccharomyces cerevisiae product, comprises actication of culture, enlarged culturing, fermentor cultivation, and fermentor cultivation finishes the back culture is separated, gets product after dry, the packing.
Wherein, actication of culture can adopt ordinary method well known to those skilled in the art to carry out, and cultivates activation as adopting slant strains.
Wherein, enlarged culturing can comprise level liquid seed culture, the cultivation of secondary liquid seeds and fermentor tank seed culture.
Its Production Flow Chart is following: slant strains cultivation → level liquid seed culture → secondary liquid seeds cultivation → fermentor tank seed culture → fermentor cultivation → separation → drying → packing.
Specifically can adopt following method:
1) slant strains is cultivated: with the Zinc-rich saccharomyces cerevisiae bacterial classification inoculation of anti-high zinc to sugar degree be 5～25%, zinc concentration is that 150～5000 μ g/ml, pH value are on 4～6.5 the wort solid inclined-plane, 26～35 ℃ of cultivations 24～48 hours.
2) level liquid seed culture: with the barms on inclined-plane inoculation 1～4 encircle to that the sugar degree that 100～500ml is housed is 5～25%, zinc concentration is that 150～5000 μ g/ml, pH value are in 4～6.5 the special-purpose culturing bottle of wort, 26～35 ℃ of cultivations 24～48 hours.
3) the secondary liquid seeds is cultivated; With the level liquid seed culture fluid be inoculated into that sugar (mixtures of wort or molasses or wort and the molasses) amount of containing that 10～40L is housed is 5～25%, zinc concentration is that 150～5000 μ g/ml, pH value are in 4～6.5 the Ka Shi culturing bottle; Inoculum size is 1%～10%, cultivates 18～48 hours at 26～35 ℃.
4) fermentor tank seed culture: the secondary seed nutrient solution is inoculated into 0.5～15m is housed
3Sugar (mixtures of wort or molasses or wort and the molasses) amount of containing be 5～25%, zinc concentration is 150～5000 μ g/ml, replenish suitable nitrogenous source and phosphorus source, pH value is in 4～6.5 the seed fermentation jar; Inoculum size is 1%～10%, cultivates 18～48 hours at 26～35 ℃.
5) ferment tank is cultivated: with the fermentor tank seed culture fluid be inoculated into that 1～300 ton sugar (mixtures of amylum hydrolysate of the sugar or molasses or amylum hydrolysate of the sugar and the molasses) amount that contains is 5～25%, zinc) concentration is that 150～5000 μ g/ml, additional suitable nitrogenous source and phosphorus source, pH value are in 4～6.5 the fermentor tank; Inoculum size is 5%～15%; Used nutrition source adopts fed-batch mode; Zinc adopts fed-batch mode to add, and cultivates 14～36 hours at 26～35 ℃.
6) separate: adopt stacked separating machine to separate again and filter or use filter press with vacuum drum.
7) drying: adopt 60～140 ℃ of fluidized dryings, the exsiccant product has activity; Adopt 100～180 ℃ of spraying dryings perhaps with the vapor pressure roller drying of 0.3～0.6Mpa, it is powdery or strip that drying prods does not have activity; Also can adopt freeze-drying dry.
8) packing: can adopt the packing of various ways to become the Zinc-rich saccharomyces cerevisiae of high zinc content.Zinc content reaches 2000～80000mg/kg.
In fermentor tank seed culture and ferment tank cultivation, need to add an amount of nitrogenous source and phosphorus source, nitrogenous source is selected from sulfate of ammoniac, bicarbonate of ammonia, ammoniacal liquor, phosphoric acid ammonia, phosphoric acid hydrogen ammonia and DAP, or the combination of these materials.The phosphorus source is for being selected from phosphoric acid, phosphoric acid ammonia, phosphoric acid hydrogen ammonia and DAP, or the combination of these materials.Be preferably phosphoric acid ammonia, phosphoric acid hydrogen ammonia and DAP.The usage quantity in preferred nitrogenous source and phosphorus source is 1～10% of substratum weight.The zinc source is selected from zinc sulfate, zinc chloride, zinc nitrate, Zinc Gluconate, amino-acid zinc etc., preferred amino acid chelating zinc, and usage quantity is 1～10% of a substratum weight, flows in 14 hours to fermenting from the fermentation beginning to add
If successive production can increase the fermentation of one grade fermemtation jar secondary seed; Fermentation condition is: the fermentor tank seed culture fluid is inoculated into that 1～300 ton sugar (mixtures of amylum hydrolysate of the sugar or molasses or amylum hydrolysate of the sugar and the molasses) amount of containing is 5～25%, zinc (zinc-amino acid chelate) concentration is 150～5000 μ g/ml, replenishes suitable nitrogenous source and phosphorus source, pH value is in 4～6.5 the secondary seed fermentor tank; Used nutrition source adopts fed-batch mode; Zinc adopts fed-batch mode to add, and cultivates 14～36 hours at 26～35 ℃.Carry out whizzer or sheet frame then and separate, in 4～10 ℃ of storage tanks or freezer, preserve, be inoculated into as required then and carry out the next stage cultivation in the fermentor tank again.This moment, the condition optimization of ferment tank was: the fermentor tank seed culture fluid is inoculated into that 1～300 ton sugar degree is 5～25%, concentration of iron is 150～5000 μ g/ml, replenishes suitable nitrogenous source and phosphorus source, pH value is in 4～6.5 the fermentor tank; Used nutrition source adopts fed-batch mode; Zinc adopts fed-batch mode to add, and cultivates 14～36 hours at 26～35 ℃.
The Zinc-rich saccharomyces cerevisiae bacterial classification of the anti-high zinc that the present invention uses, anti-zinc stable performance, growth velocity is fast, and is high to the utilization ratio of zinc, contains the zinc height and can reach 2000～80000mg/kg.And the Zinc-rich saccharomyces cerevisiae zinc content that prior art is produced is usually below 1000mg/kg, the highest 2000mg/kg that just arrives.In addition, the Zinc-rich saccharomyces cerevisiae that the present invention produces high zinc content can be the yeast viable cell, and the Zinc-rich saccharomyces cerevisiae that prior art is produced is not have active Zinc-rich saccharomyces cerevisiae mostly.And the biological zinc content of the Zinc-rich saccharomyces cerevisiae of the high zinc content that the present invention produces is high, can reach 90%.
Embodiment
Come further to set forth the present invention below in conjunction with concrete embodiment.Only if specialize, experimental technique and reagent used among the present invention are method known in those skilled in the art and reagent.Should be appreciated that these embodiment only are used to explain the present invention, and can not limit protection scope of the present invention.To those skilled in the art, under the prerequisite that does not deviate from the present invention's spirit and essence, various changes or change that the nutrient media components in these embodiments, content, culture condition, separation processing conditions are carried out also belong to protection scope of the present invention.
Embodiment 1: the present invention is to the screening of yeast saccharomyces cerevisiae bacterial classification
1) barms of the anti-high zinc of screening: yeast saccharomyces cerevisiae (Saccharomycescerevisiae) is inoculated into containing in the zinc substratum of different concns, the bacterial classification that screening can be grown in high zinc substratum.Pick out the bacterial classification of the bacterial classification of anti-zinc as domestication.
2) bacterial classification inoculation of anti-zinc is cultivated to the substratum of high density zinc, and improved zinc concentration in the substratum gradually, keep zinc in higher concentration.Pick out anti-high zinc and stable bacterial classification as the mutagenesis bacterial classification.
3) with the mutagenesis in the substratum of the lithium chloride of different concns of select bacterial classification inoculation.
4) bacterial classification with mutagenesis carries out separation screening again, utilize shake bottle ferment with the small-sized fermentation jar select zinc content height, growth velocity is fast, productive rate is high, to the barms of the high anti-high zinc of the utilization ratio of zinc, be the Zinc-rich saccharomyces cerevisiae bacterial classification.Through screening obtain that a strain zinc content is high, growth velocity is fast, productive rate is high, to the barms of the high anti-high zinc of the utilization ratio of zinc, with its called after Z1.4, and carry out preservation.
5) screening and culturing based formulas
Malt extract medium sugar degree 10% contains zinc (zinc-amino acid chelate) amount 150～5000 μ g/ml, and the pH value is 5.5.
6) screening and culturing condition
30 ℃ of temperature, stir culture 24 hours.
Embodiment 2: the production of Zinc-rich saccharomyces cerevisiae product
Produce the Production Flow Chart of high zinc content Zinc-rich saccharomyces cerevisiae: slant strains cultivation → level liquid seed culture → secondary liquid seeds cultivation → fermentor tank seed culture → fermentor cultivation → separation → drying → packing.Concrete technology is following:
1) slant strains is cultivated: with the Zinc-rich saccharomyces cerevisiae bacterial classification inoculation of anti-high zinc to sugar degree be 10%, zinc (zinc-amino acid chelate) concentration is that 150 μ g/ml, pH value are on 5.5 the wort solid inclined-plane, at 30 ℃, cultivated 24 hours.
2) level liquid seed culture: with the barms on inclined-plane inoculation 3 encircle to that sugar (wort and the molasses half and half) amount that contains that 250ml is housed is 10%, zinc (zinc-amino acid chelate) concentration is that 150 μ g/ml, pH value are in 5.5 the wort special use culturing bottle, 30 ℃ of cultivations 24 hours.
3) the secondary liquid seeds is cultivated; With the level liquid seed culture fluid be inoculated into that sugar (wort and the molasses half and half) amount of containing that 25L is housed is 10%, zinc (zinc-amino acid chelate) concentration is that 150 μ g/ml, pH value are in 5.5 the Ka Shi culturing bottle, 30 ℃ of cultivations 24 hours.
4) fermentor tank seed culture: the secondary seed nutrient solution is inoculated into sugar (wort and the molasses half and half) amount of containing that 8m3 is housed is 20%, zinc (zinc-amino acid chelate) concentration is 150 μ g/ml; Add 5% phosphoric acid hydrogen ammonia, pH value and be in 4.5 the seed fermentation jar, cultivated 48 hours at 30 ℃.
5) ferment tank is cultivated: with the fermentor tank seed culture fluid be inoculated into 150 tons contain sugar (amylum hydrolysate of the sugar and molasses half and half) measure be 20%, zinc (zinc-amino acid chelate) concentration is 150 μ g/ml, add 5% phosphoric acid hydrogen ammonia, the pH value is in 5.0 the fermentor tank; Used nutrition source adopts fed-batch mode; Zinc adopts fed-batch mode to add, and cultivates 36 hours at 20 ℃.
6) separate: adopt stacked separating machine to separate again and filter or use filter press with vacuum drum.
7) drying: adopt 100 ℃ of fluidized dryings, the exsiccant product has activity.
8) packing: can adopt the packing of various ways to become the Zinc-rich saccharomyces cerevisiae of high zinc content.Zinc content reaches 2000mg/kg.
Embodiment 3: the production of Zinc-rich saccharomyces cerevisiae product
Produce the Production Flow Chart of high zinc content Zinc-rich saccharomyces cerevisiae: slant strains cultivation → level liquid seed culture → secondary liquid seeds cultivation → fermentor tank seed culture → fermentor cultivation → separation → drying → packing.Concrete technology is following:
1) slant strains is cultivated: with the Zinc-rich saccharomyces cerevisiae bacterial classification inoculation of anti-high zinc to sugar degree be 10%, zinc (zinc-amino acid chelate) concentration is that 500 μ g/ml, pH value are on 5.5 the wort solid inclined-plane, at 30 ℃, cultivated 24 hours.
2) level liquid seed culture: with the barms on inclined-plane inoculation 4 encircle to that sugar (amylum hydrolysate of the sugar) amount that contains that 500ml is housed is 10%, zinc (zinc-amino acid chelate) concentration is that 500 μ g/ml, pH value are in 5.5 the special-purpose culturing bottle of wort, 30 ℃ of cultivations 24 hours.
3) the secondary liquid seeds is cultivated; With the level liquid seed culture fluid be inoculated into that sugar (mixtures of wort and the molasses) amount of containing that 40L is housed is 10%, zinc (zinc-amino acid chelate) concentration is that 500 μ g/ml, pH value are in 5.5 the Ka Shi culturing bottle, 30 ℃ of cultivations 24 hours.
4) fermentor tank seed culture: the secondary seed nutrient solution is inoculated into 15m is housed
3Sugar (molasses) amount of containing be 15%, zinc (zinc-amino acid chelate) concentration is 500 μ g/ml, add 1% phosphoric acid hydrogen ammonia, pH value and be in 5.5 the seed fermentation jar, 35 ℃ of cultivations 20 hours.
5) ferment tank is cultivated: with the fermentor tank seed culture fluid be inoculated into 1～300 ton contain sugar (mixtures of amylum hydrolysate of the sugar and molasses) measure be 15%, zinc (zinc-amino acid chelate) concentration is that to add 3% phosphoric acid hydrogen ammonia, pH value be in 6.0 the fermentor tank to 500 μ g/ml; Used nutrition source adopts fed-batch mode; Zinc adopts fed-batch mode to add, and cultivates 24 hours at 30 ℃.
6) separate: adopt stacked separating machine to separate again and filter or use filter press with vacuum drum.
7) drying: adopt 60～140 ℃ of fluidized dryings, the exsiccant product has activity; Adopt 100～180 ℃ of dryings of dusting perhaps with the vapor pressure roller drying of 0.3～0.6Mpa, it is powdery or strip that drying prods does not have activity; Also can adopt freeze-drying dry.
8) packing: can adopt the packing of various ways to become the Zinc-rich saccharomyces cerevisiae of high zinc content.Zinc content reaches 6700mg/kg.
Embodiment 4: the production of Zinc-rich saccharomyces cerevisiae product
Produce the Production Flow Chart of high zinc content Zinc-rich saccharomyces cerevisiae: slant strains cultivation → level liquid seed culture → secondary liquid seeds cultivation → fermentor tank seed culture → fermentor cultivation → separation → drying → packing.Concrete technology is following:
1) slant strains is cultivated: with the Zinc-rich saccharomyces cerevisiae bacterial classification inoculation of anti-high zinc to sugar degree be 10%, zinc (zinc-amino acid chelate) concentration is that 1000 μ g/ml, pH value are on 5.5 the wort solid inclined-plane, at 30 ℃, cultivated 24 hours.
2) level liquid seed culture: with the barms on inclined-plane inoculation 1～4 encircle to that sugar (wort and the molasses half and half) amount that contains that 300ml is housed is 10%, zinc (zinc-amino acid chelate) concentration is that 1000 μ g/ml, pH value are in 5.5 the wort special use culturing bottle, 30 ℃ of cultivations 24 hours.
3) the secondary liquid seeds is cultivated; With the level liquid seed culture fluid be inoculated into that sugar (wort and the molasses half and half) amount of containing that 10～40L is housed is 10%, zinc (zinc-amino acid chelate) concentration is that 1000 μ g/ml, pH value are in 5.5 the Ka Shi culturing bottle, 30 ℃ of cultivations 24 hours.
4) fermentor tank seed culture: the secondary seed nutrient solution is inoculated into 0.5～15m is housed
3Sugar (mixtures of wort or molasses or wort and the molasses) amount of containing be 5～25%, zinc (zinc-amino acid chelate) concentration is 1000 μ g/ml, add 1～5% phosphoric acid hydrogen ammonia, pH value and be in 4～6.5 the seed fermentation jar, 26～35 ℃ of cultivations 18～48 hours.
5) ferment tank is cultivated: with the fermentor tank seed culture fluid be inoculated into 1～300 ton contain sugar (mixtures of amylum hydrolysate of the sugar or molasses or amylum hydrolysate of the sugar and molasses) measure be 5～25%, zinc (zinc-amino acid chelate) concentration is 1000 μ g/ml, add 1～5% phosphoric acid hydrogen ammonia, the pH value is in 4～6.5 the fermentor tank; Used nutrition source adopts fed-batch mode; Zinc adopts fed-batch mode to add, and cultivates 14～36 hours at 26～35 ℃.
6) separate: adopt stacked separating machine to separate again and filter or use filter press with vacuum drum.
7) drying: adopt 60～140 ℃ of fluidized dryings, the exsiccant product has activity; Adopt 100～180 ℃ of dryings of dusting perhaps with the vapor pressure roller drying of 0.3～0.6Mpa, it is powdery or strip that drying prods does not have activity; Also can adopt freeze-drying dry.
8) packing: can adopt the packing of various ways to become the Zinc-rich saccharomyces cerevisiae of high zinc content.Zinc content reaches 13800mg/kg.
Embodiment 5: the production of Zinc-rich saccharomyces cerevisiae product
Produce the Production Flow Chart of high zinc content Zinc-rich saccharomyces cerevisiae: slant strains cultivation → level liquid seed culture → secondary liquid seeds cultivation → fermentor tank seed culture → fermentor cultivation → separation → drying → packing.Concrete technology is following:
1) slant strains is cultivated: with the Zinc-rich saccharomyces cerevisiae bacterial classification inoculation of anti-high zinc to sugar degree be 10%, zinc (zinc-amino acid chelate) concentration is that 5000 μ g/ml, pH value are on 5.5 the wort solid inclined-plane, at 30 ℃, cultivated 24 hours.
2) level liquid seed culture: with the barms on inclined-plane inoculation 1～4 encircle to that the sugar degree that 100～500ml is housed is 10%%, zinc (zinc-amino acid chelate) concentration is that 5000 μ g/ml, pH value are in 5.5 the special-purpose culturing bottle of wort, 30 ℃ of cultivations 24 hours.
3) the secondary liquid seeds is cultivated; With the level liquid seed culture fluid be inoculated into that sugar (mixtures of wort or molasses or wort and the molasses) amount of containing that 10～40L is housed is 10%%, zinc (zinc-amino acid chelate) concentration is that 4500 μ g/ml, pH value are in 5.5 the Ka Shi culturing bottle, 30 ℃ of cultivations 24 hours.
4) fermentor tank seed culture: the secondary seed nutrient solution is inoculated into sugar (mixtures of wort or molasses or wort and the molasses) amount of containing that 0.5～15m3 is housed is 5～25%, zinc (zinc-amino acid chelate) concentration is 4500 μ g/ml; Add 1～5% phosphoric acid hydrogen ammonia, pH value and be in 4～6.5 the seed fermentation jar, cultivated 18～48 hours at 26～35 ℃.
5) ferment tank is cultivated: with the fermentor tank seed culture fluid be inoculated into 1～300 ton contain sugar (mixtures of amylum hydrolysate of the sugar or molasses or amylum hydrolysate of the sugar and molasses) measure be 5～25%, zinc (zinc-amino acid chelate) concentration is 4500 μ g/ml, add 1～5% phosphoric acid hydrogen ammonia, the pH value is in 4～6.5 the fermentor tank; Used nutrition source adopts fed-batch mode; Zinc adopts fed-batch mode to add, and cultivates 14～36 hours at 26～35 ℃.
6) separate: adopt stacked separating machine to separate again and filter or use filter press with vacuum drum.
7) drying: adopt 60～140 ℃ of fluidized dryings, the exsiccant product has activity; Adopt 100～180 ℃ of dryings of dusting perhaps with the vapor pressure roller drying of 0.3～0.6Mpa, it is powdery or strip that drying prods does not have activity; Also can adopt freeze-drying dry.
8) packing: can adopt the packing of various ways to become the Zinc-rich saccharomyces cerevisiae of high zinc content.Zinc content reaches 80000mg/kg.
Embodiment 6: the production of Zinc-rich saccharomyces cerevisiae product
Present embodiment increases one grade fermemtation jar seed culture, and production technique is: slant strains cultivation → level liquid seed culture → secondary liquid seeds cultivation → one grade fermemtation jar seed culture → second order fermentation jar seed culture → fermentor cultivation → separation → drying → packing.The one grade fermemtation jar seed culture method and the fermentor cultivation that are increased are roughly the same; Other technologies are with embodiment 2; Just ferment tank is cultivated and is selected amylum hydrolysate of the sugar 8% and molasses to do that carbon source 5%, ammoniacal liquor 3% are made nitrogenous source, phosphoric acid hydrogen ammonia 2% is done the phosphorus source; Zinc content in the nutrient solution is 2000 μ g/ml, and the pH value is 5.2, and 28～32 ℃ of air blast of temperature were cultivated 26 hours.
The Zinc-rich saccharomyces cerevisiae zinc content of the high zinc content that present embodiment is produced reaches 25900mg/kg.
Embodiment 7: the production of Zinc-rich saccharomyces cerevisiae product
The working method of present embodiment is with embodiment 2; But in drying process, adopt fluidized exsiccant method drying to provide the Zinc-rich saccharomyces cerevisiae of activated high zinc content, drying process is: 120～140 ℃ of EATs, 70～100 ℃ of air outlet temperatures; Dry 20～50 minutes, moisture was less than 7%.
Production technique:
Slant strains cultivation → level liquid seed culture → secondary liquid seeds cultivation → fermentor tank seed culture → fermentor cultivation → separation → drying → packing.
The living cell rate of the Zinc-rich saccharomyces cerevisiae of the high zinc content that present embodiment is produced reaches more than 80%.
In sum; The Microbial Genetical Breeding technology that makes full use of of the present invention; Microbial fermentation technology; According to yeast the characteristic of zinc tolerance is filtered out the barms of anti-high zinc, tame again, induced-mutation technique selects that zinc content is high, growth velocity is fast, productive rate is high, to the Zinc-rich saccharomyces cerevisiae bacterial classification of the high anti-high zinc of the utilization ratio of zinc.Produce the Zinc-rich saccharomyces cerevisiae of high zinc content to the special process of this bacterial classification formulation.Characteristics are: anti-zinc stable performance, and growth velocity is fast, and is high to the utilization ratio of zinc; Biological zinc content is high, can reach 90%; The Zinc-rich saccharomyces cerevisiae of this high zinc content can be that activity does not have activity yet.Has more wide application space.
Claims (9)
1. a rich zinc yeast saccharomyces cerevisiae (Saccharomyces cerevisiae Hansen) Z1.4, its preserving number is CCTCC No:M 205126.
2. the working method of the said Zinc-rich saccharomyces cerevisiae of claim 1, it comprises: actication of culture, enlarged culturing and fermentation culture; Said actication of culture is that slant strains is cultivated activation; Said enlarged culturing comprises level liquid seed culture, the cultivation of secondary liquid seeds and fermentor tank seed culture; Said fermentation culture is that ferment tank is cultivated.
3. method as claimed in claim 2, its also be included in after the fermentation culture to culture separate, dry and packing.
4. like claim 2 or 3 described methods; Wherein slant strains is cultivated the activatory method and is: with said Zinc-rich saccharomyces cerevisiae bacterial classification inoculation to sugar degree be 5～25%, zinc concentration is that 150～5000 μ g/ml, pH value are on 4～6.5 the wort solid inclined-plane, 26～35 ℃ of cultivations 24～48 hours.
5. like claim 2 or 3 described methods, wherein enlarged culturing comprises:
The level liquid seed culture: with activatory bacterial classification inoculation 1～4 encircle to that the sugar degree that 100～500ml is housed is 5～25%, zinc concentration is that 150～5000 μ g/ml, pH value are in 4～6.5 the special-purpose culturing bottle of wort, cultivated 24～48 hours at 26～35 ℃;
The secondary liquid seeds is cultivated; With the level liquid seed culture fluid be inoculated into that the sugar degree that 10～40L is housed is 5～25%, zinc concentration is that 150～5000 μ g/ml, pH value are that inoculum size is 1%～10%, cultivates 18～48 hours at 26～35 ℃ in 4～6.5 the Ka Shi culturing bottle;
Fermentor tank seed culture: the secondary seed nutrient solution is inoculated into 0.5～15m is housed
3Sugar degree be 5～25%, zinc concentration is 150～5000 μ g/ml, replenish suitable nitrogenous source and phosphorus source, pH value is that inoculum size was 1%～10%, 26～35 ℃ of cultivations 18～48 hours in 4～6.5 the seed fermentation jar.
6. like claim 2 or 3 described methods; It is characterized in that; The ferment tank cultured method is: ferment tank is cultivated: the fermentor tank seed culture fluid is inoculated into that 1～300 ton sugar degree is 5～25%, zinc concentration is 150～5000 μ g/ml, replenishes suitable nitrogenous source and phosphorus source, pH value is that inoculum size is 5%～15%, used nutrition source employing fed-batch mode in 4～6.5 the fermentor tank; Zinc adopts fed-batch mode to add, and cultivates 14～36 hours at 26～35 ℃.
7. method as claimed in claim 5; It also is included in second order fermentation jar seed fermentation and cultivates; Cultural method is: the fermentor tank seed culture fluid is inoculated into that 1～300 ton sugar degree is 5～25%, zinc concentration is 150～5000 μ g/ml, replenishes suitable nitrogenous source and phosphorus source, pH value is that inoculum size is 1%～10%, used nutrition source employing fed-batch mode in 4～6.5 the fermentor tank; Zinc adopts fed-batch mode to add, and cultivates 14～36 hours at 26～35 ℃.
8. method as claimed in claim 3 is wherein separated and is adopted stacked separating machine to separate, and filters or use filter press with vacuum drum again.
9. method as claimed in claim 3, wherein, drying
A) adopt 60～140 ℃ of fluidized dryings; Or
B) adopt 100～180 ℃ of spraying dryings; Or
C) with the vapor pressure roller drying of 0.3～0.6Mpa; Or
D) adopt freeze-drying dry.
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