CN1141375C - Se-enriched high-biomass yeast and its preparing process - Google Patents
Se-enriched high-biomass yeast and its preparing process Download PDFInfo
- Publication number
- CN1141375C CN1141375C CNB011006005A CN01100600A CN1141375C CN 1141375 C CN1141375 C CN 1141375C CN B011006005 A CNB011006005 A CN B011006005A CN 01100600 A CN01100600 A CN 01100600A CN 1141375 C CN1141375 C CN 1141375C
- Authority
- CN
- China
- Prior art keywords
- selenium
- biomass
- mcg
- yeast
- yeast rich
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
- 240000004808 Saccharomyces cerevisiae Species 0.000 title claims abstract description 109
- 239000002028 Biomass Substances 0.000 title claims abstract description 70
- 238000000034 method Methods 0.000 title claims description 33
- 230000008569 process Effects 0.000 title description 2
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 claims abstract description 191
- 229910052711 selenium Inorganic materials 0.000 claims abstract description 159
- 239000011669 selenium Substances 0.000 claims abstract description 159
- 238000004519 manufacturing process Methods 0.000 claims abstract description 41
- 238000000855 fermentation Methods 0.000 claims abstract description 27
- 230000004151 fermentation Effects 0.000 claims abstract description 27
- 210000005253 yeast cell Anatomy 0.000 claims abstract description 19
- 238000005516 engineering process Methods 0.000 claims abstract description 10
- 238000012216 screening Methods 0.000 claims abstract description 10
- 238000009395 breeding Methods 0.000 claims abstract description 6
- 230000001488 breeding effect Effects 0.000 claims abstract description 6
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 98
- 239000007788 liquid Substances 0.000 claims description 56
- 210000004027 cell Anatomy 0.000 claims description 32
- 229940082569 selenite Drugs 0.000 claims description 32
- MCAHWIHFGHIESP-UHFFFAOYSA-L selenite(2-) Chemical compound [O-][Se]([O-])=O MCAHWIHFGHIESP-UHFFFAOYSA-L 0.000 claims description 32
- 239000002054 inoculum Substances 0.000 claims description 24
- 238000003756 stirring Methods 0.000 claims description 24
- 230000001580 bacterial effect Effects 0.000 claims description 18
- 230000002356 anti-selenium Effects 0.000 claims description 12
- 238000011218 seed culture Methods 0.000 claims description 11
- 238000001035 drying Methods 0.000 claims description 8
- 238000012856 packing Methods 0.000 claims description 8
- 238000010298 pulverizing process Methods 0.000 claims description 8
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 6
- 240000008042 Zea mays Species 0.000 claims description 6
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 6
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 6
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 6
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 6
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 6
- 235000005822 corn Nutrition 0.000 claims description 6
- 235000015097 nutrients Nutrition 0.000 claims description 6
- 238000009402 cross-breeding Methods 0.000 claims description 5
- 235000013379 molasses Nutrition 0.000 claims description 5
- 235000019890 Amylum Nutrition 0.000 claims description 4
- 229920002472 Starch Polymers 0.000 claims description 4
- 239000000413 hydrolysate Substances 0.000 claims description 4
- 230000004913 activation Effects 0.000 claims description 3
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 3
- 239000012531 culture fluid Substances 0.000 claims description 3
- 210000000820 garland cell Anatomy 0.000 claims description 3
- 230000002068 genetic effect Effects 0.000 claims description 3
- 230000013011 mating Effects 0.000 claims description 3
- 238000002703 mutagenesis Methods 0.000 claims description 3
- 231100000350 mutagenesis Toxicity 0.000 claims description 3
- 238000004806 packaging method and process Methods 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 239000007787 solid Substances 0.000 claims description 3
- 241000894007 species Species 0.000 claims description 3
- 238000012262 fermentative production Methods 0.000 claims description 2
- 238000005457 optimization Methods 0.000 claims description 2
- 229940081969 saccharomyces cerevisiae Drugs 0.000 claims 2
- 230000008901 benefit Effects 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 2
- 231100000252 nontoxic Toxicity 0.000 abstract description 2
- 230000003000 nontoxic effect Effects 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 8
- 238000010521 absorption reaction Methods 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 241001465754 Metazoa Species 0.000 description 4
- 230000007547 defect Effects 0.000 description 4
- 241000282412 Homo Species 0.000 description 3
- 230000008827 biological function Effects 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000004321 preservation Methods 0.000 description 3
- 235000016068 Berberis vulgaris Nutrition 0.000 description 2
- 241000335053 Beta vulgaris Species 0.000 description 2
- 101100476962 Drosophila melanogaster Sirup gene Proteins 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- GZCGUPFRVQAUEE-SLPGGIOYSA-N aldehydo-D-glucose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O GZCGUPFRVQAUEE-SLPGGIOYSA-N 0.000 description 2
- 230000001093 anti-cancer Effects 0.000 description 2
- 230000031018 biological processes and functions Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 210000000130 stem cell Anatomy 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 235000020357 syrup Nutrition 0.000 description 2
- 239000011573 trace mineral Substances 0.000 description 2
- 235000013619 trace mineral Nutrition 0.000 description 2
- 208000008316 Arsenic Poisoning Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- QLNFINLXAKOTJB-UHFFFAOYSA-N [As].[Se] Chemical compound [As].[Se] QLNFINLXAKOTJB-UHFFFAOYSA-N 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 230000002019 anti-mutation Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 229910052785 arsenic Inorganic materials 0.000 description 1
- RQNWIZPPADIBDY-UHFFFAOYSA-N arsenic atom Chemical compound [As] RQNWIZPPADIBDY-UHFFFAOYSA-N 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 1
- 229960003180 glutathione Drugs 0.000 description 1
- 235000003969 glutathione Nutrition 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
一种含有高生物量的富硒酵母及其生产方法，它是通过筛选，获得生物量高的酵母菌种和抗硒能力强及细胞硒含量高的酵母菌种，然后采用杂交育种技术，选育出含有高生物量和高硒含量的富硒酵母菌种，在发酵培养基和优化的发酵条件下，发酵培养制得含有高生物量的富硒酵母，每克干富硒酵母细胞含有机硒达500-1500微克。本发明生产的富硒酵母安全无毒，投资少、见效快、效益高。其生产方法实用性强，操作简便，易于大批量生产或因陋就简普遍推广应用。
A selenium-enriched yeast with high biomass and a production method thereof, which is to obtain yeast strains with high biomass and strong selenium resistance and high selenium content through screening, and then adopt hybrid breeding technology to select Selenium-enriched yeast strains with high biomass and high selenium content are bred, and under the fermentation medium and optimized fermentation conditions, the selenium-enriched yeast with high biomass is produced by fermentation, and each gram of dry selenium-enriched yeast cells contains organic Selenium reaches 500-1500 micrograms. The selenium-enriched yeast produced by the invention is safe and non-toxic, has low investment, quick effect and high benefit. The production method has strong practicability, simple and convenient operation, and is easy to be mass-produced or widely popularized and applied.
Description
Technical field
The present invention relates to the fermentation industry technical field, refer in particular to a kind of yeast rich in selenium and production method thereof that contains high-biomass.
Background technology
As everyone knows, selenium is the intravital a kind of essential trace element of humans and animals, it is the important component of organism glutathion inside peralcohol, many-sided biological function is arranged, wherein topmost biological function is to remove the too much active oxygen radical that produces in the organism, thereby has and improve organism immunizing power, anti-ageing, anticancer effect.In addition, environment arseniasis is a worldwide problem, and it is harmful to the health of humans and animals that its harm not only shows as recent arsenic, and even more serious is to cause that human body malignization at a specified future date changes---canceration, sudden change and distortion.According to arsenic-selenium medicine principle of toxinicide each other, selenium has and suppresses and anticancer change, anti-mutation and anti-distored biological function in addition, and the yeast that is rich in selenium acts on will bring into play significantly aspect prevention and the treatment endemic arsenic poisoning.In addition, yeast cell also contains 14 kinds of essential trace elements of rich in protein, nucleic acid, VITAMIN and body, healthy and helpful to body.
In recent years, although the existing production that is called " yeast rich in selenium ", these productions exist obvious defects and deficiency:
The first is not carried out any screening and breeding to barms, uses a kind of barms at random, owing to the resistance of different barmses to selenium has than big-difference, thus also very big to the absorption and conversion capability difference of plasma selenium.
It two is plasma selenium concentration, the culture condition of the substratum of production fermentation culture yeast rich in selenium not to be optimized, though therefore the yeast of cultivating is called " yeast rich in selenium ", but this being called,, the selenium content of " yeast rich in selenium " cell was lower, and the biomass of yeast cell is also lower.
Producing the method be called " yeast rich in selenium " cell in the prior art has three kinds substantially, and first kind is to adopt mixed processing method, and a certain amount of selenate or selenite and yeast cell are mixed.Its major defect is: it does not transform into organoselenium with inorganic selenium, the what is called that this selenate or selenite and yeast cell mix " yeast rich in selenium " can make humans and animals very low to its specific absorption, the biological action that produces is relatively poor, is not real yeast rich in selenium.
Second kind is to adopt simple adsorption method, bread yeast on the market or cerevisiae be suspended in the aqueous solution that contains selenate or selenite handle certain hour, make plasma selenium be adsorbed in yeast cell surface, its major defect is: this processing still can not transform into organoselenium with inorganic selenium, the yeast cell of this absorption selenate or selenite and above-mentioned first method do not have the difference of essence, can make the human or animal very low to its specific absorption, the biological action that produces is relatively poor, is not real yeast rich in selenium equally.
The third is a culturing yeast cell in containing the substratum of plasma selenium, but barms is not carried out any screening and breeding, use a kind of barms at random, its major defect is: owing to the resistance of different barmses to selenium has than big-difference, thus widely different to the absorption and conversion capability of plasma selenium.In addition, this technology is not optimized plasma selenium concentration, the culture condition of substratum, therefore adopt in the yeast of this technology cultivation, organic selenium content is lower, and organoselenium 500 micrograms are following, the also low economic benefit of cellular biomass is also very low thereby every gram dry yeast cell contains.
Summary of the invention
The object of the present invention is to provide a kind of yeast rich in selenium and production method thereof that contains high-biomass, utilize modern high-new biotechnology, physio-biochemical characteristics according to different barmses, by screening, obtain high barms and the high barms of cell selenium content of biomass, the yeast rich in selenium bacterial classification that adopts crossbreeding technology to select then to contain high-biomass and high selenium content, reaching the fermentation condition bottom fermentation in the substratum of optimizing cultivates, by suitable production technique, acquisition contains the yeast rich in selenium product of high-biomass, and the dried yeast rich in selenium cell of every gram contains organoselenium and reaches the 500-1500 microgram.
The object of the present invention is achieved like this: a kind of yeast rich in selenium that contains high-biomass, it is characterized in that: it is by screening, make the high barms of biomass and anti-selenium ability reaches the high barms of cell selenium content by force, adopt crossbreeding technology then, select the yeast rich in selenium bacterial classification that contains high-biomass and high selenium content, fermentation culture obtains to contain the yeast rich in selenium of high-biomass and high selenium content in the substratum of optimizing, the dried yeast rich in selenium cell of every gram contains organoselenium and reaches the 500-1500 microgram, classification called after: yeast saccharomyces cerevisiae (yeast rich in selenium) Saccharomyces cerevisiae, preserving number is: CGMCC NO.05 39, preservation day is: January 15 calendar year 2001.
The present invention also provides a kind of production method that contains the yeast rich in selenium of high-biomass, and it is characterized in that: it comprises the steps:
1), preparation contains the yeast rich in selenium bacterial classification of high-biomass: obtain to contain the yeast rich in selenium excellent species of high-biomass and high selenium content by the means of molecular breeding, its method that adopts is:
1. according to the physio-biochemical characteristics of different barmses, in containing the substratum of different selenium concentrations, screen the high barms of biomass respectively and anti-selenium ability reaches the high barms of cell selenium content by force;
2. the high barms of biomass and anti-selenium ability are reached by force that the high barms of cell selenium content is given birth to spore, monoploid separates, the monoploid mutant strain that has different mating types and different genetic markers through mutagenesis screening;
3. the biomass that is separated to is high yeast monoploid mutant strain and anti-selenium ability reach the high yeast monoploid mutant strain of cell selenium content by force and hybridize;
4. at the diploid hybrid strain excellent of selecting to screen on the substratum high-biomass and high selenium content, i.e. yeast rich in selenium bacterial classification;
2), fermentation culture contains the yeast rich in selenium of high-biomass and high selenium content:
(1), fermentative medium formula:
The wort of 8%～20% total reducing sugar amount of selection includes the selenate of 200～400 mcg/ml or the selenite of 400～600 mcg/ml, and the pH of substratum is 4.5-7.0;
(2), fermentation culture conditions: the culture condition that fermentation culture contains the yeast rich in selenium of high-biomass and high selenium content is: the inoculum size of seed liquor is 5-30%, under 25-32 ℃ condition, stir culture 16-30 hour;
3), produce the yeast rich in selenium that contains high-biomass and high selenium content: its production technique comprises: slant strains → liquid spawn → level liquid seed culture → secondary liquid seeds is cultivated → three grades of liquid seeds cultivation → ferment tank cultivation → collection yeast cell and drying → pulverizing and packing, and its each production technique is as follows:
(1), slant strains: the yeast rich in selenium bacterial classification is inoculated in that to contain total sugar content be that 8-20%, selenate concentration are 200～400 mcg/ml or use selenite, its concentration is on the wort solid inclined-plane of 400～600 mcg/ml,, put into 4 ℃ of refrigerators and preserve after 24-72 hour 25-32 ℃ of cultivation;
(2), liquid spawn: after the yeast rich in selenium bacterial classification activation of preserving, connecing a garland cells is that 8-20%, selenate concentration are 200～400 mcg/ml or selenite in 150 milliliters of total sugar contents are housed, its concentration is in the triangular flask of malt extract medium of 400～600 mcg/ml, in 25-32 ℃ of stir culture 14-20 hour, be liquid spawn;
(3), level liquid seed culture: it is that 8-20%, selenate concentration are 200～400 mcg/ml or selenite that liquid spawn is equipped with 150 milliliters of total sugar contents by 5～20% 30 of inoculum size accesses, its concentration is in the triangular flask of malt extract medium of 400～600 mcg/ml, at 25-32 ℃ of stir culture 14-20 hour, be the level liquid inoculum;
(4), the secondary liquid seeds is cultivated: it is that 8-20%, selenate concentration are 200～400 mcg/ml or use selenite that the first order seed nutrient solution is equipped with 45 liters of total sugar contents by 5～20% inoculum size access, its concentration is the Xiao Kashi fermentor tank of the malt extract medium of 400～600 mcg/ml, at 25-32 ℃ of stir culture 14-20 hour, be secondary liquid seeds culture;
(5), three grades of liquid seeds are cultivated: it is that 8-20%, selenate concentration are 200～600 mcg/ml or use selenite that the secondary seed nutrient solution is equipped with 450 liters of total sugar contents by 10～30% inoculum size access, its concentration is the Da Kashi fermentor tank of the malt extract medium of 400～600 mcg/ml, at 25-32 ℃ of stir culture 14-20 hour, be three grades of liquid seeds cultures;
(6), ferment tank is cultivated: it is that 8-20%, selenate concentration are 200～400 mcg/ml or use selenite that three grades of seed culture fluids are equipped with 2250 liters of total sugar contents by 20～40% inoculum size access, its concentration was the fermentor tank of the malt extract medium of 400～600 mcg/ml, at 25-32 ℃ of stir culture 14-24 hour;
(7), collect yeast cell and drying: adopt sheet frame squeezing or centrifugal collection yeast rich in selenium cell.Air-dry under 45～85 ℃ of conditions, moisture is less than 10%;
(8), pulverize: with pulverizer with air-dry yeast rich in selenium cell pulverization;
(9), packing: use air-locked packaging bags, be the yeast rich in selenium of the selenium-enriched content of high-biomass, the dried yeast rich in selenium cell of every gram contains organoselenium and reaches the 500-1500 microgram, classification called after: yeast saccharomyces cerevisiae (yeast rich in selenium) Saccharomyces cerevisiae, preserving number is: CGMCC NO.0539, preservation day is: January 15 calendar year 2001.
This fermentative medium formula and fermentation culture conditions are: the total reducing sugar amount is 12% hydrolysis sugar, 0.8% corn hydrolyzed solution, 0.5% ammonium sulfate, the selenite of the selenate of 300 mcg/ml or 500 mcg/ml, the pH of substratum is 5.0, inoculum size is 15%, under 28-30 ℃ condition, and stir culture 30 hours.This fermentative medium formula and fermentation culture conditions are: the molasses of total reducing sugar amount 15%, 1.0% corn hydrolyzed solution, 0.5% ammonium sulfate, 0.1% phosphoric acid, the selenite of the selenate of 350 mcg/ml or 450 mcg/ml, the pH of substratum was 6.0, and inoculum size is 20%, 26-28 ℃ of stir culture 24 hours.This production technique that contains the yeast rich in selenium of high-biomass and high selenium content comprises the cultivation of level Four seed liquor.This production technique that contains the yeast rich in selenium of high-biomass and high selenium content comprises the variation of doing suitable increase or minimizing technology with actual production conditions.
Major advantage of the present invention is as follows:
1, the yeast rich in selenium bacterial classification of the present invention's use, not only has high-biomass, and the organic selenium content of cell is higher, and its organic selenium content contains the 500-1500 microgram up to every gram stem cell, and the organic selenium content of the product that prior art is produced is that every gram stem cell contains below organoselenium 500 micrograms.
2, the fermentation substrate that uses of method of the present invention is common wort or makes fermentation substrate with amylum hydrolysate of the sugar or molasses, only needs to regulate its total sugar content, adds selenate or selenite, and method is simple, and is with low cost.
3, method of the present invention does not have particular requirement to fermentation equipment and condition, and the equipment and the condition of general fermentation plant all can be produced, and the production investment is less.
4, the yeast rich in selenium product safety of method production of the present invention is nontoxic, less investment, instant effect, high efficiency.
5, method of the present invention is practical, and is easy and simple to handle, is easy to generally promote.
6, press the biomass height of the yeast rich in selenium product of method production of the present invention, the organic selenium content height of yeast cell, quality is good, and the dried yeast rich in selenium cell of every gram contains organoselenium and reaches the 500-1500 microgram.
Further specify below in conjunction with preferred embodiment and accompanying drawing.
Description of drawings
Fig. 1 is a technological process of production synoptic diagram of the present invention.
Embodiment
Embodiment 1
Consult Fig. 1, the production method that contains the yeast rich in selenium of high-biomass of the present invention comprises the steps:
1), preparation contains the yeast rich in selenium bacterial classification of high-biomass: because occurring in nature does not have biomass and the simultaneously all high barms of cell selenium content, can only could obtain to contain the yeast rich in selenium excellent species of high-biomass and high selenium content by the means of molecular breeding.The method that the present invention adopts is:
1. according to the physio-biochemical characteristics of different barmses, in containing the substratum of different selenium concentrations, screen the high barms of biomass respectively and anti-selenium ability reaches the high barms of cell selenium content by force.
2. the high barms of biomass and anti-selenium ability are reached by force that the high barms of cell selenium content is given birth to spore, monoploid separates, the monoploid mutant strain that has different mating types and different genetic markers through mutagenesis screening.
3. high yeast monoploid mutant strain and the anti-selenium ability of biomass that is separated to reached the high yeast monoploid mutant strain cross-breeding of cell selenium content by force.
4. at the diploid hybrid strain excellent of selecting to screen on the substratum high-biomass and high selenium content, i.e. yeast rich in selenium bacterial classification.
2), cultivate the yeast rich in selenium that contains high-biomass and high selenium content in the fermentation culture conditions top fermentation of fermention medium and optimization:
(1), fermentative medium formula:
The wort of 8%～20% total reducing sugar amount of selection includes the selenate of 200～400 mcg/ml or the selenite of 400～600 mcg/ml, and the pH of substratum is 4.5-7.0.
(2), the fermentation culture conditions of You Huaing:
The culture condition that fermentative production contains the yeast rich in selenium of high-biomass and high selenium content is: the inoculum size of yeast rich in selenium bacterial classification is 5-30%, under 25-32 ℃ of condition, and stir culture 16-30 hour;
3), produce the yeast rich in selenium product that contains high-biomass and high selenium content: its production technique comprises:
Slant strains → liquid spawn → level liquid seed culture → secondary liquid seeds is cultivated the yeast rich in selenium of the selenium-enriched content of → three grades of liquid seeds cultivation → ferment tank cultivation → collection yeast cell and drying → pulverizing and packing high-biomass.Further specify as follows to each production technique below:
(1), slant strains: the yeast rich in selenium bacterial classification is inoculated in that to contain total sugar content be that 8-20%, selenate concentration are that 200～400 mcg/ml (or are used selenite, its concentration is 400～600 mcg/ml) wort solid inclined-plane on, under 25-32 ℃ condition, cultivate after 24-72 hour, put into 4 ℃ of refrigerators and preserve.
(2), liquid spawn: after the yeast rich in selenium bacterial classification activation of preserving, connecing a garland cells is that 8-20%, selenate concentration are that 200～400 mcg/ml (or are used selenite in 150 milliliters of total sugar contents are housed, its concentration is 400～600 mcg/ml) the triangular flask of malt extract medium in, under 25-32 ℃ condition, stir culture 14-20 hour, be liquid spawn.
(3), level liquid seed culture: it is that 8-20%, selenate concentration are that 400～600 mcg/ml (or are used selenite that liquid spawn is equipped with 150 milliliters of total sugar contents by 5～20% inoculum size access, its concentration is 400～600 mcg/ml) the triangular flask of malt extract medium in, under 25-32 ℃ condition, stir culture 14-20 hour, be the level liquid inoculum.
(4), the secondary liquid seeds is cultivated: it is that 8-20%, selenate concentration are that 200～400 mcg/ml (or are used selenite that the first order seed nutrient solution is equipped with 45 liters of total sugar contents by 5～20% inoculum size access, its concentration is 400～600 mcg/ml) the Xiao Kashi fermentor tank of malt extract medium, under 25-32 ℃ condition, stir culture 14-20 hour, be secondary liquid seeds culture.
(5), three grades of liquid seeds are cultivated: it is that 8-20%, selenate concentration are that 200～400 mcg/ml (or are used selenite that the secondary seed nutrient solution is equipped with 450 liters of total sugar contents by 10～30% inoculum size access, its concentration is 400～600 mcg/ml) the Da Kashi fermentor tank of malt extract medium, under 25-32 ℃ condition, stir culture 14-20 hour, be three grades of liquid seeds cultures.
(6), ferment tank is cultivated: it is that 8-20%, selenate concentration are that 200～400 mcg/ml (or are used selenite that three grades of seed culture fluids are equipped with 2250 liters of total sugar contents by 20～40% inoculum size access, its concentration is 400～600 mcg/ml) the fermentor tank of malt extract medium, under 25-32 ℃ condition, stir culture 14-24 hour.
(7), collect yeast cell and drying: adopt sheet frame squeezing or centrifugal collection yeast rich in selenium cell.Air-dry under 45～85 ℃ of conditions, moisture is less than 10%.
(8), pulverize: with pulverizer with air-dry yeast rich in selenium cell pulverization.
(9), packing: use air-locked packaging bags, be the yeast rich in selenium of the selenium-enriched content of high-biomass, the dried yeast rich in selenium cell of every gram contains organoselenium and reaches the 500-1500 microgram, classification called after: yeast saccharomyces cerevisiae (yeast rich in selenium) Saccharomyces cerevisiae, preserving number is: CGMCC NO.0539, preservation day is: January 15 calendar year 2001.
Production method of the present invention can be according to the fermentation equipment and the condition of producer, and the adjustment that above technology is done suitably to increase or reduce changes.
Embodiment 2
Production method of the present invention can be selected different fermentative medium formulas, culture condition and contain high-biomass and the suitable adjustment change that increases or reduce of the production technique of the yeast rich in selenium of high selenium content do according to the different material of different areas.
Present embodiment is that selection is a substratum with the amylum hydrolysate of the sugar, its fermentative medium formula and culture condition are: the total reducing sugar amount is 12% amylum hydrolysate of the sugar, 0.8% corn hydrolyzed solution, 0.5% ammonium sulfate, the selenite of the selenate of 350 mcg/ml or 550 mcg/ml, the pH of substratum are 5.0, and the inoculum size of seed liquor is 15%, under 28-30 ℃ condition, stir culture 30 hours.
Its production technique comprises: slant strains → liquid spawn → level liquid seed culture → secondary liquid seeds is cultivated the yeast rich in selenium of → three grades of liquid seeds cultivation → level Four liquid seeds cultivation → ferment tank cultivation → collection yeast cell and drying → pulverizing and the high selenium content of packing high-biomass.The dried yeast rich in selenium cell of every gram contains organoselenium and reaches 1000 micrograms.
The production technique of present embodiment increases the level Four liquid seeds cultivates, and its condition is identical substantially with three grades of liquid seeds cultivations, and other processing condition are substantially the same manner as Example 1, so do not repeat.
Embodiment 3
Present embodiment is that selection is a substratum with sucrose molasses or beet sirup, its fermentative medium formula and culture condition are: the total reducing sugar amount is 15% sucrose molasses or beet sirup, 1.0% corn hydrolyzed solution, 0.8% ammonium sulfate, 0.1% phosphoric acid, the selenite of the selenate of 300 mcg/ml or 500 mcg/ml, the pH of substratum is 6.0, and the inoculum size of seed liquor is 20%, 26-28 ℃ of stir culture 28 hours.
Its production technique comprises: liquid spawn → level liquid seed culture → secondary liquid seeds is cultivated the yeast rich in selenium of → three grades of liquid seeds cultivation → level Four liquid seeds cultivation → Pyatyi liquid seeds cultivation → ferment tank cultivation → collection yeast cell and drying → pulverizing and the high selenium content of packing high-biomass.The dried yeast rich in selenium cell of every gram contains organoselenium and reaches 1200 micrograms.
The production technique of present embodiment increases the Pyatyi liquid seeds cultivates, and its condition is identical substantially with three grades of liquid seeds cultivations, and other processing condition are substantially the same manner as Example 2, so do not repeat.
In sum, method of the present invention is to make full use of microbial technique, physio-biochemical characteristics according to different barmses, by screening, obtain high barms and the anti-selenium ability of biomass and reach the high barms of cell selenium content by force, the yeast rich in selenium bacterial classification that adopts crossbreeding technology to select then to contain high-biomass and high selenium content, fermentation culture in the fermention medium of optimizing, acquisition contains the yeast rich in selenium of high-biomass and high selenium content, and it is low that this yeast rich in selenium that contains high-biomass and high selenium content has a cost, raw material is extensive, easy and simple to handle, fermentation period is short, characteristics such as the biomass height of yeast cell and organic selenium content height.Adopt production yeast rich in selenium method of the present invention, be fit to large-scale production and the production of short run on the basis of the existing conditions no matter how simple and crude, have actual application prospect widely, and have important social benefit and huge economic benefit.
Claims (6)
1, a kind of yeast rich in selenium that contains high-biomass, it is characterized in that: it is by screening, obtain high barms and the anti-selenium ability of biomass and reach the high barms of cell selenium content by force, adopt crossbreeding technology then, select the yeast rich in selenium bacterial classification that contains high-biomass and high selenium content, under the fermentation condition of fermention medium and optimization, fermentation culture production contains the yeast rich in selenium of high-biomass, and the dried yeast rich in selenium cell of every gram contains organoselenium and reaches the 500-1500 microgram; The classification called after: yeast saccharomyces cerevisiae (Saccharomycescerevisiae), preserving number is: CGMCC NO.0539.
2, a kind of production method that contains the yeast rich in selenium of high-biomass, it is characterized in that: it comprises the steps:
1), preparation contains the yeast rich in selenium bacterial classification of high-biomass: obtain to contain the yeast rich in selenium excellent species of high-biomass and high selenium content by the means of molecular breeding, its method that adopts is:
1. according to the physio-biochemical characteristics of different barmses, in containing the substratum of different selenium concentrations, screen the high barms of biomass respectively and anti-selenium ability reaches the high barms of cell selenium content by force;
2. the high barms of biomass and anti-selenium ability are reached by force that the high barms of cell selenium content is given birth to spore, monoploid separates, the monoploid mutant strain that has different mating types and different genetic markers through mutagenesis screening;
3. the biomass that is separated to is high yeast monoploid mutant strain and anti-selenium ability reach the high yeast monoploid mutant strain of cell selenium content by force and hybridize;
4. at the diploid hybrid strain excellent of selecting to screen on the substratum high-biomass and high selenium content, i.e. yeast rich in selenium bacterial classification;
2), fermentation culture contains the yeast rich in selenium of high-biomass and high selenium content:
(1), fermentative medium formula: select the wort of 8%～20% total reducing sugar amount, contain the selenate of 200～400 mcg/ml or the selenite of 400～600 mcg/ml, the pH of substratum is 4.5-7.0;
(2), fermentation culture conditions: the culture condition that fermentative production contains the yeast rich in selenium of high-biomass and high selenium content is: the seed liquor inoculum size is 5-30%, under 25-32 ℃ condition, stir culture 16-30 hour;
3), produce the yeast rich in selenium that contains high-biomass and high selenium content: its production technique in turn includes the following steps: slant strains, liquid spawn, level liquid seed culture, secondary liquid seeds are cultivated, three grades of liquid seeds cultivations, ferment tanks are cultivated, collected yeast cell and drying, pulverizing and packing, and its each production technique is as follows:
(1), slant strains: the yeast rich in selenium bacterial classification is inoculated in that to contain total sugar content be that 8-20%, selenate concentration are 200～400 mcg/ml or use selenite, its concentration is on the wort solid inclined-plane of 400～600 mcg/ml,, put into 4 ℃ of refrigerators and preserve after 24-72 hour 25-32 ℃ of cultivation;
(2), liquid spawn: after the yeast rich in selenium bacterial classification activation of preserving, connecing a garland cells is that 8-20%, selenate concentration are 200～400 mcg/ml or use selenite in 150 milliliters of total sugar contents are housed, its concentration was in the triangular flask of malt extract medium of 400～800 mcg/ml, was liquid spawn in 25-32 ℃ of stir culture 14-20 hour;
(3), level liquid seed culture: it is that 8-20%, selenate concentration are 200～400 mcg/ml or use selenite that liquid spawn is equipped with 150 milliliters of total sugar contents by 5～20% 30 of inoculum size accesses, its concentration is in the triangular flask of malt extract medium of 400～600 mcg/ml, at 25-32 ℃ of stir culture 14-20 hour, be the level liquid inoculum;
(4), the secondary liquid seeds is cultivated: it is that 8-20%, selenate concentration are 200～400 mcg/ml or use selenite that the first order seed nutrient solution is equipped with 45 liters of total sugar contents by 5～20% inoculum size access, its concentration is the Xiao Kashi fermentor tank of the malt extract medium of 400～600 mcg/ml, at 25-32 ℃ of stir culture 14-20 hour, be secondary liquid seeds culture;
(5), three grades of liquid seeds are cultivated: it is that 8-20%, selenate concentration are 200～400 mcg/ml or use selenite that the secondary seed nutrient solution is equipped with 450 liters of total sugar contents by 10～30% inoculum size access, its concentration is the Da Kashi fermentor tank of the malt extract medium of 400～600 mcg/ml, at 25-32 ℃ of stir culture 14-20 hour, be three grades of liquid seeds cultures;
(6), ferment tank is cultivated: it is that 8-20%, selenate concentration are 200～400 mcg/ml or use selenite that three grades of seed culture fluids are equipped with 2250 liters of total sugar contents by 20～40% inoculum size access, its concentration was the fermentor tank of the malt extract medium of 400～600 mcg/ml, at 25-32 ℃ of stir culture 14-24 hour;
(7), collect yeast cell and drying: adopt sheet frame squeezing or centrifugal collection yeast rich in selenium cell.Air-dry under 45～85 ℃ of conditions, moisture is less than 10%;
(8), pulverize: with pulverizer with air-dry yeast rich in selenium cell pulverization;
(9), packing: use air-locked packaging bags, be the yeast rich in selenium that contains high-biomass, the dried yeast rich in selenium cell of every gram contains organoselenium and reaches the 500-1500 microgram, and the classification called after: yeast saccharomyces cerevisiae (Saccharomyces cerevisiae), preserving number is: CGMCC NO.0539.
3, production method as claimed in claim 2, it is characterized in that: this fermentative medium formula and culture condition are: the total reducing sugar amount is 12% amylum hydrolysate of the sugar, 0.8% corn hydrolyzed solution, 0.5% ammonium sulfate, the selenite of the selenate of 350 mcg/ml or 550 mcg/ml, the pH of substratum are 5.0, and the inoculum size of seed liquor is 15%, under 28-30 ℃ condition, stir culture 30 hours.
4, production method as claimed in claim 2, it is characterized in that: the prescription and the culture condition of this fermention medium are: the total reducing sugar amount is 15% molasses, 1.0% corn hydrolyzed solution, 0.5% ammonium sulfate, 0.1% phosphoric acid, the selenite of the selenate of 300 mcg/ml or 500 mcg/ml, the pH of substratum was 6.0, and the inoculum size of seed liquor is 20%, 26-28 ℃ of stir culture 24 hours.
5, production method as claimed in claim 2 is characterized in that: this production technique that contains the yeast rich in selenium of high-biomass comprises that increasing the level Four liquid seeds cultivates.
6, production method as claimed in claim 5 is characterized in that: this production technique that contains the yeast rich in selenium of high-biomass comprises that increasing the Pyatyi liquid seeds cultivates.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011006005A CN1141375C (en) | 2001-01-17 | 2001-01-17 | Se-enriched high-biomass yeast and its preparing process |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011006005A CN1141375C (en) | 2001-01-17 | 2001-01-17 | Se-enriched high-biomass yeast and its preparing process |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1309177A CN1309177A (en) | 2001-08-22 |
| CN1141375C true CN1141375C (en) | 2004-03-10 |
Family
ID=4651722
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB011006005A Expired - Lifetime CN1141375C (en) | 2001-01-17 | 2001-01-17 | Se-enriched high-biomass yeast and its preparing process |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1141375C (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103911299A (en) * | 2012-12-31 | 2014-07-09 | 安琪酵母股份有限公司 | Selenium-rich yeast and preparation method thereof |
| CN108192854A (en) * | 2018-01-31 | 2018-06-22 | 浙江大学 | A kind of richness manganese yeast product and its production method |
Families Citing this family (24)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100347286C (en) * | 2004-04-16 | 2007-11-07 | 中国科学院微生物研究所 | Calcium enriched beer yeast strain and use |
| CN100451098C (en) * | 2006-03-30 | 2009-01-14 | 安琪酵母股份有限公司 | Selenium-rich and chromium-rich Saccharomyces cerevisiae, selenium-rich and chromium-rich yeast product and their production process |
| CN100451099C (en) * | 2006-03-30 | 2009-01-14 | 安琪酵母股份有限公司 | Selenium-rich saccharomyces cerevisiae, selenium-rich yeast product and their production process |
| CN100390270C (en) * | 2006-07-21 | 2008-05-28 | 江南大学 | A kind of preparation method of selenium-enriched edible mushroom product with activated selenium ore as selenium source |
| CN102181371B (en) * | 2011-02-25 | 2013-06-26 | 浙江深友生物技术有限公司 | Yeast and method for producing selenium-enriched yeast by utilizing yeast |
| CN102511305A (en) * | 2011-12-07 | 2012-06-27 | 苏州硒谷科技有限公司 | Method for cultivating high-selenium-tolerability strain of edible fungi |
| CN102703561B (en) * | 2012-05-26 | 2014-03-19 | 彭祚全 | Method for producing activated selenium mineral powder by microbial fermentation |
| CN102703513A (en) * | 2012-05-26 | 2012-10-03 | 彭祚全 | Method for preparing red elemental selenium by utilizing super-selenium resistance microorganism |
| CN103315220B (en) * | 2013-05-08 | 2014-11-12 | 浙江国曲生物科技有限公司 | Selenium-rich monascus production method |
| CN103329994B (en) * | 2013-07-11 | 2014-10-29 | 谷劲松 | Production method of probiotic active milk beverage rich in organic selenium |
| US10592064B2 (en) | 2013-09-17 | 2020-03-17 | Amazon Technologies, Inc. | Approaches for three-dimensional object display used in content navigation |
| CN103667086B (en) * | 2013-12-04 | 2019-08-13 | 北京大北农科技集团股份有限公司 | One plant height biomass Se-enriched yeast, additive and premix containing it |
| CN104543399B (en) * | 2015-01-20 | 2018-05-08 | 河南科技大学 | A kind of preparation method of yeast selenium forage additive |
| CN105602798A (en) * | 2016-03-13 | 2016-05-25 | 青岛嘉瑞生物技术有限公司 | Selenium-rich Salicornia europaea L. health care liquor and production process |
| CN106244477B (en) * | 2016-10-12 | 2020-01-10 | 厦门元尊生物工程有限公司 | Selenium-rich saccharomyces cerevisiae strain and application thereof in preparation of selenium-rich purple sweet potato yeast product |
| CN106906206A (en) * | 2017-04-26 | 2017-06-30 | 中国科学院合肥物质科学研究院 | A kind of Se-enriched yeast, preparation method and application |
| CN107594070A (en) * | 2017-09-08 | 2018-01-19 | 四川亿客农食品有限公司 | A kind of breeding method of Se-rich farm products and its application |
| CN108633717A (en) * | 2018-08-13 | 2018-10-12 | 重庆善硒科技有限公司 | A kind of selenium-enriched broccoli soilless culture nutrient fluid formula and its cultural method |
| CN109232107A (en) * | 2018-11-19 | 2019-01-18 | 黑龙江牧康牧业有限责任公司 | Yeast selenium foliar fertilizer, production method and application |
| CN109370930A (en) * | 2018-12-06 | 2019-02-22 | 安琪酵母(德宏)有限公司 | A kind of Se-enriched yeast and preparation method thereof |
| CN110250464A (en) * | 2019-07-17 | 2019-09-20 | 武汉市鑫宏食品酿造科研所 | A kind of preparation method of the Selenium-enriched fermentation phoenix head salty base of ginger |
| CN113025510B (en) * | 2019-12-25 | 2022-09-16 | 中国科学院微生物研究所 | Function-enhanced yeast culture rich in organic trace elements and preparation method thereof |
| CN113322223B (en) * | 2021-06-03 | 2023-09-12 | 重庆市畜牧科学院 | Selenium-rich yeast genetically engineered bacteria and their surface display systems and construction methods |
| CN114794303B (en) * | 2022-05-10 | 2023-07-14 | 吉林省农业科学院 | A kind of selenium-enriched red yeast gum, feed additive for selenium-enriched red yeast gum and its preparation method and application |
-
2001
- 2001-01-17 CN CNB011006005A patent/CN1141375C/en not_active Expired - Lifetime
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103911299A (en) * | 2012-12-31 | 2014-07-09 | 安琪酵母股份有限公司 | Selenium-rich yeast and preparation method thereof |
| CN103911299B (en) * | 2012-12-31 | 2016-02-10 | 安琪酵母股份有限公司 | Yeast rich in selenium and preparation method thereof |
| CN108192854A (en) * | 2018-01-31 | 2018-06-22 | 浙江大学 | A kind of richness manganese yeast product and its production method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1309177A (en) | 2001-08-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1141375C (en) | Se-enriched high-biomass yeast and its preparing process | |
| CN1273004C (en) | Method for producing hedgehog fungus and special culture medium thereof | |
| CN101914478B (en) | Bacillus subtilis and application thereof | |
| CN108991318B (en) | Seaweed fermented beverage | |
| CN103667086A (en) | Selenium-enriched yeast with high biomass, and additive and premix containing the same | |
| CN1919998A (en) | Process for preparing winter jujube wine | |
| CN103820299A (en) | Worm grass mycelium fermented vinegar and preparation method thereof | |
| CN102351580B (en) | Method for preparing black fungus nutrient solution by utilizing food processing residues | |
| CN1561666A (en) | Method for preparing tremella mesenterica cultivating seed | |
| CN114107073A (en) | A kind of method that utilizes molasses to produce mycelin | |
| CN1142266C (en) | Cr-enriched yeast containing high biomass and its preparing process | |
| CN1759760A (en) | Method for producing fruit juice of germinant unpolished rice and ganoderma lucidum, and production | |
| CN1024468C (en) | Method for preparing mycoprotein feed by solid fermentation of mixed crude starch material | |
| CN112442453B (en) | Ormokodak yeast for degrading biogenic amine and application of same in white spirit brewing | |
| CN1313596C (en) | High-biomass zinc-riched yeast, breeding method and use thereof | |
| CN109694829A (en) | A kind of selenium-rich saccharomyces cerevisiae, selenium-rich richness lycopene saccharomyces cerevisiae and preparation method thereof | |
| CN107550946A (en) | A kind of preparation and application of poultry immunity reinforcing agent | |
| CN1264969C (en) | Zymogen agent for treating agricultural wastes and preparation method | |
| CN1141377C (en) | Saccharomyces cerevisiae with high trehalose content and production method thereof | |
| CN115039633B (en) | A method for artificially cultivating the fruiting body of Corymborhinus japonicus | |
| CN1498962A (en) | Aspergillus tubingensis capable of high pectinase production and its application in solid-state fermentation | |
| KR101810567B1 (en) | Method for producing dry yeast containing selenium using fermentation | |
| CN1177925C (en) | Fermenting culture process of produicng Cordyceps extract | |
| CN1242068C (en) | Method for producing somatic powder containing lovastatin by liquid submerged fermentation | |
| CN100338211C (en) | Agricultural waste fermenting bacterial prepn and its prepn process |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C06 | Publication | ||
| PB01 | Publication | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20090109 Address after: Anhui Province Jieshou City Zhongyuan Road No. 439 Patentee after: Anhui Huaxin Natural Medicine Co., Ltd. Address before: No. 13, North Zhongguancun, Beijing, Haidian District Patentee before: Institute of Microbiology, Chinese Academy of Sciences |
|
| ASS | Succession or assignment of patent right |
Owner name: ANHUI PROVINCE HUAXIN BIOLOGY PHARMACY CO., LTD. Free format text: FORMER OWNER: INST OF MICROBES, CHINESE ACADEMY OF SCIENCES Effective date: 20090109 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Se-enriched high-biomass yeast and its preparing process Effective date of registration: 20150911 Granted publication date: 20040310 Pledgee: The development of small and medium-sized enterprises financing Company Limited by Guarantee Jieshou City Pledgor: Anhui Huaxin Natural Medicine Co., Ltd. Registration number: 2015990000772 |
|
| PLDC | Enforcement, change and cancellation of contracts on pledge of patent right or utility model | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20161020 Granted publication date: 20040310 Pledgee: The development of small and medium-sized enterprises financing Company Limited by Guarantee Jieshou City Pledgor: Anhui Huaxin Natural Medicine Co., Ltd. Registration number: 2015990000772 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Se-enriched high-biomass yeast and its preparing process Effective date of registration: 20161020 Granted publication date: 20040310 Pledgee: The development of small and medium-sized enterprises financing Company Limited by Guarantee Jieshou City Pledgor: Anhui Huaxin Natural Medicine Co., Ltd. Registration number: 2016990000886 |
|
| PLDC | Enforcement, change and cancellation of contracts on pledge of patent right or utility model | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20171211 Granted publication date: 20040310 Pledgee: The development of small and medium-sized enterprises financing Company Limited by Guarantee Jieshou City Pledgor: Anhui Huaxin Natural Medicine Co., Ltd. Registration number: 2016990000886 |
|
| CX01 | Expiry of patent term | ||
| CX01 | Expiry of patent term |
Granted publication date: 20040310 |