CN100451099C - Selenium-rich saccharomyces cerevisiae, selenium-rich yeast product and their production process - Google Patents
Selenium-rich saccharomyces cerevisiae, selenium-rich yeast product and their production process Download PDFInfo
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Abstract
The present invention relates to one kind of selenium-rich Saccharomyces cerevisiae, selenium-rich Saccharomyces cerevisiae product and their production process. Of the selenium-rich Saccharomyces cerevisiae product, each kilogram contains selenium in 4000-8000 mg, and organoselenium accounts for 90-99 wt% of the total selenium content. The process of producing selenium-rich Saccharomyces cerevisiae product includes the following steps: 1. fermenting the selenium-rich Saccharomyces cerevisiae of the present invention in culture medium containing selenium in 10-300 mcg/ml at 26-35 deg.c; and 2. drying the fermented Saccharomyces cerevisiae liquid to obtain the selenium-rich Saccharomyces cerevisiae product. The selenium-rich Saccharomyces cerevisiae product is safe and non-toxic, and may be used in medicine, food, feed and other fields.
Description
Technical field
The present invention relates to a kind of rich selenium yeast saccharomyces cerevisiae, derive from the yeast rich in selenium product and the production method thereof of this richness selenium yeast saccharomyces cerevisiae.
Background technology
Selenium is a kind of essential trace element of humans and animals, is the important component part of plurality of enzymes, and selenium is the Selenoperoxidase important component, and by redox, the protection microbial film is avoided infringement, keeps the normal function of cell.Selenium can also play strengthening immunity, protection cardiovascular, promote growth, antitumor, detoxifcation, regulate function such as metabolism.The endemy that causes at the scarce selenium of China has Keshan disease and Kaschin-Beck disease.Yeast rich in selenium is obvious for replenishing the some diseases effect that causes because of scarce selenium, and very rich nutrient contents is arranged in the yeast in addition, can bring into play synergy.
Many to yeast rich in selenium research both at home and abroad in recent years, but seldom can produce greatly, main problem is: (1) selenium content is lower, and is generally below 1000mg/kg, high at 1500mg/kg; (2) inorganic selenium content is low, and it is to be simple yeast and selenium absorption that part is arranged, and selenium methionine content is lower; (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; (4) utilization ratio to selenium is low.
Summary of the invention
The object of the invention is to provide a kind of yeast rich in selenium and production method thereof of high selenium content.Utilize GENERALIZATION OF MODERN BREEDING TECHNIQUE, movable inorganic selenium had the barms of tolerance by screening, then by the chemomorphosis technology, select the selenium content height, growth velocity is fast, productive rate is high, to the yeast rich in selenium bacterial classification of the high anti-high selenium of the utilization ratio of selenium.By the zymotechnique of optimization, obtain the yeast rich in selenium product of high selenium content again, selenium content is 1000～8000mg/kg.The yeast rich in selenium product of high selenium content is divided into to be had activity and not to have active two kinds.And, produce the yeast rich in selenium product of high selenium content at this strain bacterial classification formulation special production technology, selenium content is 1000～8000mg/kg.
Of the present invention aspect first in, a kind of rich selenium yeast saccharomyces cerevisiae Z1.2 (Saccharomycescerevisiae Hansen Z1.2) is provided, its on October 25th, 2005 at China typical culture collection center (CCTCC, Wuhan University, postcode 430072) carried out biological preservation, deposit number is CCTCC M 205124.It is to come out containing on the substratum of selenium seed selection by nitrosoguanidine mutagenesis.
In aspect second of the present invention, provide a kind of yeast rich in selenium product, it derives from the rich selenium yeast saccharomyces cerevisiae of above-mentioned the present invention, contains selenium 1000～8000mg in wherein every kg yeast rich in selenium product, and wherein organic selenium content accounts for 90～96.9 weight % of total selenium.
In a preferred embodiment aspect this, contain selenium 1000～3000mg in every kg yeast rich in selenium product of the present invention.
In a further preferred embodiment, organic selenium content accounts for the 93-96.9 weight % of total selenium in the yeast rich in selenium product of the present invention.
In another preferred embodiment of the present invention, 30～65 weight % are Sethotope in total selenium of yeast rich in selenium product of the present invention.
In bright another preferred embodiment of we, the living yeast rate in the yeast rich in selenium product of the present invention is 80～95%.
In aspect the 3rd of the present invention, provide a kind of method of production selenium-rich Saccharomyces cerevisiae product, comprised the following steps:
(a) make rich selenium yeast saccharomyces cerevisiae of the present invention in the substratum that contains selenium 10～300 μ g/ml, under 26～35 ℃ temperature, ferment;
(b), promptly get the yeast rich in selenium product with the yeast fermentation broth drying.
In a preferred embodiment of the inventive method, selenium content is 100 μ g/ml in the wherein said substratum.
In another preferred embodiment of the inventive method, wherein said leavening temperature is 30 ℃.
In another preferred embodiment of the inventive method, wherein said drying is with yeast fermentation broth dust drying or vapor pressure roller drying.
In another preferred embodiment of the inventive method, wherein said drying is that yeast fermentation broth is carried out lyophilize.
Principal feature of the present invention has:
(1) the anti-selenium stable performance of yeast rich in selenium bacterial classification of the anti-high selenium of the present invention's use, growth velocity is fast, to the utilization ratio height of selenium, contains the selenium height and can reach 8000mg/kg, and the yeast rich in selenium selenium content that former method and technology is produced is below 1000mg/kg, the highest 1600mg/kg that just arrives.
(2) the present invention's yeast rich in selenium of producing high selenium content can have activity, and the yeast rich in selenium that former technology is produced is not have active yeast rich in selenium mostly.
(3) the yeast rich in selenium organic selenium content height of the high selenium content of the present invention's production can reach 99 weight %, and wherein Sethotope content can account for 65 weight % of total selenium.
Embodiment
Of the present invention aspect first in, a kind of rich selenium yeast saccharomyces cerevisiae Z1.2 (Saccharomycescerevisiae Hansen Z1.2) is provided, its on October 25th, 2005 at China typical culture collection center (CCTCC, Wuhan University, postcode 430072) carried out biological preservation, deposit number is CCTCC M 205124.It is to come out containing on the substratum of selenium seed selection by nitrosoguanidine mutagenesis.
Inorganic selenium toxicity is bigger, general yeast is difficult under the condition of high selenium grows, even growth velocity and also not high to the utilization ratio of selenium is arranged, the present invention is by nitrosoguanidine mutagenesis, select the selenium content height, growth velocity is fast, productive rate is high, to the yeast rich in selenium bacterial classification of the high anti-high selenium of the utilization ratio of selenium.Concrete implementation step is as follows:
What 1) will be inoculated into different concns contains in the seleno culture medium bacterial classification that screening can be grown in high seleno culture medium.Pick out the bacterial classification of the bacterial classification of anti-selenium as mutagenesis.
2) bacterial classification inoculation with anti-selenium carries out mutagenesis to the substratum of the nitrosoguanidine that contains various dose.
3) yeast strain with mutagenesis carries out separation screening again, utilization shake the bottle and the small-sized fermentation jar ferment select contain selenium content height on the substratum of selenium, growth velocity is fast, productive rate is high, to the yeast rich in selenium bacterial classification of the high anti-high selenium of the utilization ratio of selenium, be the yeast rich in selenium bacterial classification of anti-high selenium.
4) screening and culturing based formulas
Malt extract medium sugar degree 5～25 weight % contain selenium (selenate or selenite) amount 10～300 μ g/ml, and the pH value is 4～6.5.
5) screening and culturing condition
26～35 ℃ of temperature, stir culture 18～48 hours.
In aspect second of the present invention, provide a kind of yeast rich in selenium product, it derives from above-mentioned rich selenium yeast saccharomyces cerevisiae, contains selenium 1000～8000mg in wherein every kg yeast rich in selenium product, preferably contains selenium 1000～3000mg; Wherein organic selenium content accounts for 90～96.9 weight % of total selenium, preferably accounts for the 93-96.9 weight % of total selenium; Further preferably, 30～65 weight % are Sethotope in total selenium of yeast rich in selenium product of the present invention, and living yeast rate (the living yeast number accounts for the per-cent of total cell number) is calculated as 80～95% through flat board dyeing.
In aspect the 3rd of the present invention, the invention provides the method for production selenium-rich Saccharomyces cerevisiae product, its Production Flow Chart is as follows: slant strains cultivation → level liquid seed culture → secondary liquid seeds cultivation → fermentor tank seed culture → fermentor cultivation → separation → drying → packing.
1) slant strains is cultivated: it is that 5～25 weight %, selenium (selenate or selenite) concentration are that 10～300 μ g/ml, pH value are on 4～6.5 the wort solid inclined-plane, 26～35 ℃ of cultivations 24～48 hours that the yeast rich in selenium bacterial classification of anti-high selenium is inoculated into sugar degree.
2) level liquid seed culture: it is that 5～25 weight %, selenium (selenate or selenite) concentration are that 10～300 μ g/ml, pH value are in 4～6.5 the special-purpose culturing bottle of wort, 26～35 ℃ of cultivations 24～48 hours that the barms on inclined-plane inoculation 1～4 is encircled to the sugar degree that 100～500ml is housed.
3) the secondary liquid seeds is cultivated: it is that 5～25 weight %, selenium (selenate or selenite) concentration are that 10～300 μ g/ml, pH value are in 4～6.5 the Ka Shi culturing bottle, 26～35 ℃ of cultivations 18～48 hours that the level liquid seed culture fluid is inoculated into sugar (mixtures of wort or molasses or wort and the molasses) amount that contains that 10～40L is housed.
4) fermentor tank seed culture: the secondary seed nutrient solution is inoculated into 1～15m is housed
3Sugar (mixtures of wort or molasses or wort and the molasses) amount of containing be that 5～25 weight %, selenium (selenate or selenite) concentration are 10～300 μ g/ml, to replenish suitable nitrogenous source and phosphorus source, pH value be in 4～6.5 the seed fermentation jar, 26～35 ℃ of cultivations 18～48 hours.
5) ferment tank is cultivated: the fermentor tank seed culture fluid is inoculated into 15～180m
3Sugar (mixtures of amylum hydrolysate of the sugar or molasses or amylum hydrolysate of the sugar and the molasses) amount of containing be that 5～25 weight %, selenium (selenate or selenite) concentration are 10～300 μ g/ml, to replenish suitable nitrogenous source and phosphorus source, pH value be in 4～6.5 the fermentor tank, used nutrition source adopts fed-batch mode, selenium adds in batches, cultivates 14～36 hours at 26～35 ℃.
6) separate: adopt stacked separating machine to separate, filter or use filter press with vacuum drum again.
7) drying: adopt 60～140 ℃ of boiling type dryings, the exsiccant product has activity; Adopt 100～180 ℃ of dryings or with the vapor pressure roller drying of 3～6Kpa, it is powdery or strip that drying products does not have activity of dusting; Also can adopt the freeze-drying drying.
8) packing: can adopt the packing of various ways to become the yeast rich in selenium product of high selenium content.
Need to add an amount of nitrogenous source and phosphorus source in fermentor tank seed culture and ferment tank cultivation, 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.
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 15～180m
3Sugar (mixtures of amylum hydrolysate of the sugar or molasses or amylum hydrolysate of the sugar and the molasses) amount of containing be that 5～25 weight %, selenium (selenate or selenite) concentration are 10～300 μ g/ml, to replenish suitable nitrogenous source and phosphorus source, pH value be in 4～6.5 the secondary seed fermentor tank, used nutrition source adopts fed-batch mode, selenium adds in batches, 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.
Can produce yeast rich in selenium product of the present invention by aforesaid method, contain selenium 1000～8000mg in wherein every kg yeast rich in selenium product, preferably contain selenium 1000～3000mg; Wherein organic selenium content accounts for 90～99 weight % of total selenium, preferably accounts for the 93-99 weight % of total selenium; Further preferably, 30～65 weight % are Sethotope in total selenium of yeast rich in selenium product of the present invention, and the living yeast rate is 80～95%.
To describe feature of the present invention and all respects in detail by specific embodiment below.Unless specialize, experimental technique and reagent used among the present invention are method known in those skilled in the art and reagent.In addition, embodiment is interpreted as illustrative, rather than will limit the scope of the invention, and the spirit and scope of the invention only are defined by the claims.To those skilled in the art, under the prerequisite that does not deviate from essence of the present invention and scope, 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 screening of yeast saccharomyces cerevisiae bacterial classification of the present invention
1) the multiple barms different with purposes of will originating is inoculated into containing in the seleno culture medium of different concns, the bacterial classification that screening can be grown in high seleno culture medium.Pick out the bacterial classification of the bacterial classification of anti-selenium as mutagenesis.
2) bacterial classification inoculation with anti-selenium carried out mutagenesis 24 hours to the substratum of the nitrosoguanidine that contains various dose.
3) yeast strain with mutagenesis carries out separation screening again, utilization shake the bottle and the small-sized fermentation jar ferment select contain selenium content height on the substratum of selenium, growth velocity is fast, productive rate is high, to the yeast rich in selenium bacterial classification of the high anti-high selenium of the utilization ratio of selenium, be the yeast rich in selenium bacterial classification S205 of anti-high selenium.
4) screening and culturing based formulas
Malt extract medium sugar degree 5～25 weight % contain selenium (selenate or selenite) amount 10～300 μ g/ml, and the pH value is 4～6.5.
5) screening and culturing condition
26～35 ℃ of temperature, stir culture 18～48 hours.
Embodiment 2: the production-scheme 1 of yeast rich in selenium product
The Production Flow Chart of high selenium content yeast rich in selenium: slant strains cultivation → level liquid seed culture → secondary liquid seeds cultivation → fermentor tank seed culture → fermentor cultivation → separation → drying → packing.Concrete technology is as follows:
1) slant strains is cultivated: it is that 5～25 weight %, selenium (selenate or selenite) concentration are that 10 μ g/ml, pH value are on 4～6.5 the wort solid inclined-plane, 26～35 ℃ of cultivations 24～48 hours that the yeast rich in selenium bacterial classification of anti-high selenium is inoculated into sugar degree.
2) level liquid seed culture: it is that 5～25 weight %, selenium (selenate or selenite) concentration are that 10 μ g/ml, pH value are in 4～6.5 the special-purpose culturing bottle of wort, 26～35 ℃ of cultivations 24～48 hours that the barms on inclined-plane inoculation 1～4 is encircled to the sugar degree that 100～500ml is housed.
3) the secondary liquid seeds is cultivated: it is that 5～25 weight %, selenium (selenate or selenite) concentration are that 10 μ g/ml, pH value are in 4～6.5 the Ka Shi culturing bottle, 26～35 ℃ of cultivations 18～48 hours that the level liquid seed culture fluid is inoculated into sugar (mixtures of wort or molasses or wort and the molasses) amount that contains that 10～40L is housed.
4) fermentor tank seed culture: the secondary seed nutrient solution is inoculated into 1～15m is housed
3Sugar (mixtures of wort or molasses or wort and the molasses) amount of containing be that 5～25 weight %, selenium (selenate or selenite) concentration are 10 μ g/ml, to add 1～5 weight % phosphoric acid hydrogen ammonia, pH value be in 4～6.5 the seed fermentation jar, 26～35 ℃ of cultivations 18～48 hours.
5) ferment tank is cultivated: the fermentor tank seed culture fluid is inoculated into 15～180m
3Sugar (mixtures of amylum hydrolysate of the sugar or molasses or amylum hydrolysate of the sugar and the molasses) amount of containing be that 5～25 weight %, selenium (selenate or selenite) concentration are 10 μ g/ml, to add 1～5 weight % phosphoric acid hydrogen ammonia, pH value be in 4～6.5 the fermentor tank, used nutrition source adopts fed-batch mode, selenium adds in batches, cultivates 14～36 hours at 26～35 ℃.
6) separate: adopt stacked separating machine to separate, filter or use filter press with vacuum drum again.
7) drying: adopt 60～140 ℃ of boiling type dryings, the exsiccant product has activity; Adopt 100～180 ℃ of dryings or with the vapor pressure roller drying of 3～6Kpa, it is powdery or strip that drying products does not have activity of dusting; Also can adopt the freeze-drying drying.
8) packing: can adopt the packing of various ways to become the yeast rich in selenium product of high selenium content.Selenium content reaches 1200mg/kg, and wherein organic selenium content accounts for 92.6 weight % of total selenium, and Sethotope content accounts for 44.3 weight % of total selenium.
Embodiment 3: the production-scheme 2 of yeast rich in selenium product
The Production Flow Chart of high selenium content yeast rich in selenium: slant strains cultivation → level liquid seed culture → secondary liquid seeds cultivation → fermentor tank seed culture → fermentor cultivation → separation → drying → packing.Concrete technology is as follows:
1) slant strains is cultivated: it is that 5～25 weight %, selenium (selenate or selenite) concentration are that 300 μ g/ml, pH value are on 4～6.5 the wort solid inclined-plane, 26～35 ℃ of cultivations 24～48 hours that the yeast rich in selenium bacterial classification of anti-high selenium is inoculated into sugar degree.
2) level liquid seed culture: it is that 5～25 weight %, selenium (selenate or selenite) concentration are that 300 μ g/ml, pH value are in 4～6.5 the special-purpose culturing bottle of wort, 26～35 ℃ of cultivations 24～48 hours that the barms on inclined-plane inoculation 1～4 is encircled to the sugar degree that 100～500ml is housed.
3) the secondary liquid seeds is cultivated: it is that 5～25 weight %, selenium (selenate or selenite) concentration are that 300 μ g/ml, pH value are in 4～6.5 the Ka Shi culturing bottle, 26～35 ℃ of cultivations 18～48 hours that the level liquid seed culture fluid is inoculated into sugar (mixtures of wort or molasses or wort and the molasses) amount that contains that 10～40L is housed.
4) fermentor tank seed culture: the secondary seed nutrient solution is inoculated into 1～15m is housed
3Sugar (mixtures of wort or molasses or wort and the molasses) amount of containing be that 5～25 weight %, selenium (selenate or selenite) concentration are 300 μ g/ml, to add 1～5 weight % phosphoric acid hydrogen ammonia, pH value be in 4～6.5 the seed fermentation jar, 26～35 ℃ of cultivations 18～48 hours.
5) ferment tank is cultivated: the fermentor tank seed culture fluid is inoculated into 15～180m
3Sugar (mixtures of amylum hydrolysate of the sugar or molasses or amylum hydrolysate of the sugar and the molasses) amount of containing be that 5～25 weight %, selenium (selenate or selenite) concentration are 300 μ g/ml, to add 1～5 weight % phosphoric acid hydrogen ammonia, pH value be in 4～6.5 the fermentor tank, used nutrition source adopts fed-batch mode, selenium adds in batches, cultivates 14～36 hours at 26～35 ℃.
6) separate: adopt stacked separating machine to separate, filter or use filter press with vacuum drum again.
7) drying: adopt 60～140 ℃ of boiling type dryings, the exsiccant product has activity; Adopt 100～180 ℃ of dryings or with the vapor pressure roller drying of 3～6Kpa, it is powdery or strip that drying products does not have activity of dusting; Also can adopt the freeze-drying drying.
8) packing: can adopt the packing of various ways to become the yeast rich in selenium product of high selenium content.Selenium content reaches 8000mg/kg, and wherein organic selenium content accounts for 91.3 weight % of total selenium, and Sethotope content accounts for 36.2 weight % of total selenium.
Embodiment 4: the production-scheme 3 of yeast rich in selenium product
The Production Flow Chart of high selenium content yeast rich in selenium: slant strains cultivation → level liquid seed culture → secondary liquid seeds cultivation → fermentor tank seed culture → fermentor cultivation → separation → drying → packing.Concrete technology is as follows:
1) slant strains is cultivated: it is that 5～25 weight %, selenium (selenate or selenite) concentration are that 100 μ g/ml, pH value are on 4～6.5 the wort solid inclined-plane, 26～35 ℃ of cultivations 24～48 hours that the yeast rich in selenium bacterial classification of anti-high selenium is inoculated into sugar degree.
2) level liquid seed culture: it is that 5～25 weight %, selenium (selenate or selenite) concentration are that 100 μ g/ml, pH value are in 4～6.5 the special-purpose culturing bottle of wort, 26～35 ℃ of cultivations 24～48 hours that the barms on inclined-plane inoculation 1～4 is encircled to the sugar degree that 100～500ml is housed.
3) the secondary liquid seeds is cultivated: it is that 5～25 weight %, selenium (selenate or selenite) concentration are that 100 μ g/ml, pH value are in 4～6.5 the Ka Shi culturing bottle, 26～35 ℃ of cultivations 18～48 hours that the level liquid seed culture fluid is inoculated into sugar (mixtures of wort or molasses or wort and the molasses) amount that contains that 10～40L is housed.
4) fermentor tank seed culture: the secondary seed nutrient solution is inoculated into 1～15m is housed
3Sugar (mixtures of wort or molasses or wort and the molasses) amount of containing be that 5～25 weight %, selenium (selenate or selenite) concentration are 100 μ g/ml, to add 1～5 weight % phosphoric acid hydrogen ammonia, pH value be in 4～6.5 the seed fermentation jar, 26～35 ℃ of cultivations 18～48 hours.
5) ferment tank is cultivated: the fermentor tank seed culture fluid is inoculated into 15～180m
3Sugar (mixtures of amylum hydrolysate of the sugar or molasses or amylum hydrolysate of the sugar and the molasses) amount of containing be that 5～25 weight %, selenium (selenate or selenite) concentration are 100 μ g/ml, to add 1～5 weight % phosphoric acid hydrogen ammonia, pH value be in 4～6.5 the fermentor tank, used nutrition source adopts fed-batch mode, selenium adds in batches, cultivates 14～36 hours at 26～35 ℃.
6) separate: adopt stacked separating machine to separate, filter or use filter press with vacuum drum again.
7) drying: adopt 60～140 ℃ of boiling type dryings, the exsiccant product has activity; Adopt 100～180 ℃ of dryings or with the vapor pressure roller drying of 3～6Kpa, it is powdery or strip that drying products does not have activity of dusting; Also can adopt the freeze-drying drying.
8) packing: can adopt the packing of various ways to become the yeast rich in selenium product of high selenium content.Selenium content reaches 4000mg/kg, and wherein organic selenium content accounts for 93.7 weight % of total selenium, and Sethotope content accounts for 58 weight % of total selenium.
Embodiment 5: the production-scheme 4 of yeast rich in selenium product
The Production Flow Chart of high selenium content yeast rich in selenium: slant strains cultivation → level liquid seed culture → secondary liquid seeds cultivation → fermentor tank seed culture → fermentor cultivation → separation → drying → packing.Concrete technology is as follows:
1) slant strains is cultivated: it is that 5～25 weight %, selenium (selenate or selenite) concentration are that 200 μ g/ml, pH value are on 4～6.5 the wort solid inclined-plane, 26～35 ℃ of cultivations 24～48 hours that the yeast rich in selenium bacterial classification of anti-high selenium is inoculated into sugar degree.
2) level liquid seed culture: it is that 5～25 weight %, selenium (selenate or selenite) concentration are that 200 μ g/ml, pH value are in 4～6.5 the special-purpose culturing bottle of wort, 26～35 ℃ of cultivations 24～48 hours that the barms on inclined-plane inoculation 1～4 is encircled to the sugar degree that 100～500ml is housed.
3) the secondary liquid seeds is cultivated: it is that 5～25 weight %, selenium (selenate or selenite) concentration are that 200 μ g/ml, pH value are in 4～6.5 the Ka Shi culturing bottle, 26～35 ℃ of cultivations 18～48 hours that the level liquid seed culture fluid is inoculated into sugar (mixtures of wort or molasses or wort and the molasses) amount that contains that 10～40L is housed.
4) fermentor tank seed culture: the secondary seed nutrient solution is inoculated into 1～15m is housed
3Sugar (mixtures of wort or molasses or wort and the molasses) amount of containing be that 5～25 weight %, selenium (selenate or selenite) concentration are 200 μ g/ml, to add 1～5 weight % phosphoric acid hydrogen ammonia, pH value be in 4～6.5 the seed fermentation jar, 26～35 ℃ of cultivations 18～48 hours.
5) ferment tank is cultivated: the fermentor tank seed culture fluid is inoculated into 15～180m
3Sugar (mixtures of amylum hydrolysate of the sugar or molasses or amylum hydrolysate of the sugar and the molasses) amount of containing be that 5～25 weight %, selenium (selenate or selenite) concentration are 200 μ g/ml, to add 1～5 weight % phosphoric acid hydrogen ammonia, pH value be in 4～6.5 the fermentor tank, used nutrition source adopts fed-batch mode, selenium adds in batches, cultivates 14～36 hours at 26～35 ℃.
6) separate: adopt stacked separating machine to separate, filter or use filter press with vacuum drum again.
7) drying: adopt 60～140 ℃ of boiling type dryings, the exsiccant product has activity; Adopt 100～180 ℃ of dryings or with the vapor pressure roller drying of 3～6Kpa, it is powdery or strip that drying products does not have activity of dusting; Also can adopt the freeze-drying drying.
8) packing: can adopt the packing of various ways to become the yeast rich in selenium product of high selenium content.Selenium content reaches 6000mg/kg, and wherein organic selenium content accounts for 96.9 weight % of total selenium, and Sethotope content accounts for 65 weight % of total selenium.
Embodiment 6: the production-scheme 5 of yeast rich in selenium 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 weight % and molasses 5 weight % to do that carbon source, ammoniacal liquor 3% are made nitrogenous source, phosphoric acid hydrogen ammonia 2% is done the phosphorus source, selenium content in the nutrient solution is 100 μ g/ml, the pH value is 5.2, and 28～32 ℃ of air blast of temperature were cultivated 26 hours.
Selenium content reaches 2500mg/kg in every kg yeast rich in selenium product of the high selenium content that this embodiment is produced.
Embodiment 7: produce the yeast rich in selenium product with active high selenium content
This embodiment adopts the method drying of fluidized drying to provide the yeast rich in selenium product of activated high selenium content, and drying process is: 120～140 ℃ of inlet temperature, and 70～100 ℃ of air outlet temperatures, dry 20～50 minutes, moisture was less than 7 weight %.
Production technique is 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 yeast rich in selenium product of the high selenium content that this embodiment is produced reaches more than 80%.Other technologies are with embodiment 2.
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 selenium tolerance is filtered out the barms of anti-high selenium, again by induced-mutation technique select the selenium content height, growth velocity is fast, productive rate is high, to the yeast rich in selenium bacterial classification of the high anti-high selenium of the utilization ratio of selenium.The special process of formulating at this bacterial classification is produced the yeast rich in selenium of high selenium content, and characteristics are: anti-selenium stable performance, and growth velocity is fast, to the utilization ratio height of selenium; The organic selenium content height can reach 99 weight %, and wherein Sethotope content can account for 65 weight % of total selenium; The yeast rich in selenium of this high selenium content can be that activity does not have activity yet.Has more wide application space.
Claims (10)
1. a rich selenium yeast saccharomyces cerevisiae Z1.2 (Saccharomyces cerevisiae Hansen Z1.2), its deposit number is CCTCC M 205124.
2. yeast rich in selenium product, it derives from the described rich selenium yeast saccharomyces cerevisiae of claim 1, contains selenium 1000～8000mg in wherein every kg yeast rich in selenium product, and wherein organic selenium content accounts for 90～96.9 weight % of total selenium.
3. the described yeast rich in selenium product of claim 2 contains selenium 1000～3000mg in wherein every kg yeast rich in selenium product.
4. the described yeast rich in selenium product of claim 2, wherein organic selenium content accounts for the 93-96.9 weight % of total selenium.
5. the described yeast rich in selenium product of claim 2,30～65 weight % are Sethotope in wherein said total selenium.
6. according to any described yeast rich in selenium product among the claim 2-5, wherein the living yeast rate is 80～95%.
7. a method of producing the described yeast rich in selenium product of claim 2 comprises the following steps:
(a) make the described rich selenium yeast saccharomyces cerevisiae of claim 1 in the substratum that contains selenium 10～300 μ g/ml, under 26～35 ℃ temperature, ferment;
(b), promptly get the yeast rich in selenium product with the yeast fermentation broth drying.
8. the described method of claim 7, selenium content is 100 μ g/ml in the wherein said substratum.
9. the described method of claim 7, wherein said leavening temperature is 30 ℃.
10. any described method among the claim 7-9, wherein said drying are with yeast fermentation broth dust drying, vapor pressure roller drying or lyophilize.
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| CN104276892A (en) * | 2014-10-13 | 2015-01-14 | 湖南省林业科学院 | Special selenium-rich liquid fertilizer for moso bamboo shoots and preparation method of fertilizer |
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