CN102533684A - Method for separating acetaldehyde dehydrogenase from Xuefeng dried yeasts - Google Patents
Method for separating acetaldehyde dehydrogenase from Xuefeng dried yeasts Download PDFInfo
- Publication number
- CN102533684A CN102533684A CN201210005420XA CN201210005420A CN102533684A CN 102533684 A CN102533684 A CN 102533684A CN 201210005420X A CN201210005420X A CN 201210005420XA CN 201210005420 A CN201210005420 A CN 201210005420A CN 102533684 A CN102533684 A CN 102533684A
- Authority
- CN
- China
- Prior art keywords
- preparation
- time
- aldh
- acetaldehyde dehydrogenase
- yeast
- 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.)
- Pending
Links
Abstract
The invention discloses a preparation method for separating acetaldehyde dehydrogenase from Xuefeng dried yeasts, which comprises the following steps: adding the Xuefeng dried yeasts to water and incubating to obtain wet yeasts; adding the wet yeasts to a phosphate buffer solution to prepare a bacterium suspension; performing ultrasonic disruption on the bacterium suspension; then, centrifuging; collecting supernate to obtain crude enzyme liquor; adding ammonium sulfate of which the saturation is 30-60 percent to the obtained crude enzyme liquor; centrifuging; collecting supernate; then, adding ammonium sulfate of which the saturation is 65-85 percent to the supernate; centrifuging; collecting sediments; adding the phosphate buffer solution to the sediments; and separating after the sediments are completely dissolved to obtain the required acetaldehyde dehydrogenase. The preparation method for separating the acetaldehyde dehydrogenase from the Xuefeng dried yeasts is capable of breaking through the limitation of the source of the acetaldehyde dehydrogenase and realizing wide source, low price and easiness in culture of the raw material, and has the advantages of high product activity, high product purity and simple and controllable reaction process.
Description
Technical field
The present invention relates to the preparing technical field of acetaldehyde dehydrogenase in the yeast, specifically is a kind of method of from the snowy peak dry yeast, separating acetaldehyde dehydrogenase.
Background technology
Acetaldehyde dehydrogenase (Aldehyde dehydrogenase ALDH) extensively is present in eukaryote and the prokaryotic organism, under the condition that nicotinamide adenine dinucleotide exists, and the dehydrogenation reaction of some aldehyde of its catalysis and ketone.In human and other many animal bodies, the plastosome acetaldehyde dehydrogenase can be the deleterious aldehydes of organism is transformed.The ethanol that human body is taken in by intestinal absorption is mainly at the liver intracellular metabolite.After ethanol gets in the body; At first under the effect of ethanol dehydrogenase (ADH), make alcohol dehydrogenase be converted into acetaldehyde; Under the katalysis by acetaldehyde dehydrogenase (ALDH), make the oxidized generation acetate of acetaldehyde then, acetate is decomposed into CO through tricarboxylic acid cycle (TCA)
2And water, and the generation ATP that releases energy.In the human body, a kind of enzyme before generally all existing, and also quantity equates basically.But the people who lacks a kind of enzyme in back is just many, and in the area, Asia, 50% people lacks ALDH.ALDH lacks, and makes ethanol be not decomposed into water and CO fully
2, continue to stay in the body with the form of acetaldehyde.Acetaldehyde is the very strong material of toxicity, can form the acetaldehyde protein complex by the covalent attachment microsomal protein, suppresses the discharge of liver cell synthetic proteins, finally causes the generation of human body alcoholic liver disease.And acetaldehyde also changes ultra oxygen compound into through XOD in vivo, causes the membrane lipid peroxo-to generate mda and nonene, and they can further promote the liver cell releasing hormone, destroy cytolemma.Accumulate in vivo when too much when acetaldehyde, the people produced after drinking blush, feel sick, symptoms such as vomiting, panhidrosis, jerk, can cause shock when serious.Acetaldehyde is the potential carcinogenic substance in vivo, can cause oral cavity, esophagus, enteron aisle, liver cancer type in the body.Therefore, acetaldehyde oxygenolysis under the katalysis of ALDH is most important.ALDH is owing to exist special katalysis, and the application in social production and daily life is with increasingly extensive.
In the prior art, the problem that preparation ALDH runs into is with grinding and chromatographic column technology separation and Extraction from animal and plant cells, and expense is high, the purifying productive rate is low, ALDH content is few, is difficult to scale operation.In another problem of running into of preparation ALDH is that existing commodity ALDH mainly is a separation and Extraction liver, pancreas or the liver cell mitochondria from animal, and resource-constrained and costing an arm and a leg has greatly hindered the using value of this product.
Summary of the invention
The objective of the invention is to deficiency, a kind of method of from the snowy peak dry yeast, separating acetaldehyde dehydrogenase is provided to prior art.This method uses the ultrasonic disruption method to come broken snowy peak dry yeast cell to obtain crude enzyme liquid ALDH; And with ammonium sulfate precipitation and Sephadex G-75 separation and purification crude enzyme liquid ALDH; Can obtain that the purifying multiple is higher, vigor is big, the ALDH of MWD about 57kDa; Products obtained therefrom can be applied to be specially adapted to the exploitation of disintoxicating product in the middle of normal food production, medical field and the genetically engineered.
Realize that the technical scheme that the object of the invention adopted comprises:
A kind of preparation method who from the snowy peak dry yeast, separates acetaldehyde dehydrogenase comprises the steps:
(1) the snowy peak dry yeast is added mixing in the entry, in 30 ℃ ~ 40 ℃ water-bath, hatch 10min ~ 20min, get deposition again behind the high speed centrifugation and promptly get the yeast that wets; Wherein the volume ratio of the weight of snowy peak dry yeast and water is: 5% ~ 15%;
(2) will wet that to add concentration be that 0.05mol/L, pH are mixed with bacteria suspension in 7.5 ~ 8.5 the phosphate buffered saline buffer to yeast, centrifugal behind this bacteria suspension ultrasonic disruption, collect supernatant, be crude enzyme liquid; The condition of its ultrasonic disruption is: treatment capacity be 15mL ~ 25mL, 1 ℃ ~ 8 ℃ of temperature, ultrasonic power be 250W ~ 350W, each ultrasonic time 1 ~ 4s, pitch time 1 ~ 8s, total ultrasonic time 10 ~ 20min; The volume ratio of wet zymic weight and phosphate buffered saline buffer is 15% ~ 25%;
(3) be that 30% ~ 60% ammonium sulfate adds in the crude enzyme liquid of above-mentioned gained with saturation ratio, centrifugal, get its supernatant; Be that 65% ~ 85% ammonium sulfate adds in this supernatant with saturation ratio again, centrifugal, get its deposition; And in this deposition, add phosphate buffered saline buffer; After treating that deposition is dissolved fully, separate, can obtain required acetaldehyde dehydrogenase enzyme.
The weight of snowy peak dry yeast and the volume ratio of water are 8% ~ 12% in the said step (1).
The bath temperature that the snowy peak dry yeast is hatched in the said step (1) is 33 ℃ ~ 37 ℃, and incubation time is 13min ~ 17min.
The volume ratio of wet zymic weight and phosphoric acid buffer is 18% ~ 22% in the said step (2).
The treatment capacity of said ultrasonic disruption is 18mL ~ 22mL.
The pH value of said phosphoric acid buffer is 7.8 ~ 8.2.
The temperature of said ultrasonic disruption is 2 ℃ ~ 6 ℃.
The power of said ultrasonic disruption is 280W ~ 320W.
The time of said each ultrasonic disruption is 1 ~ 3s, pitch time to be that 1 ~ 5s, total time are 12 ~ 18min.
Crude enzyme liquid is respectively 35% ~ 55% and 70% ~ 80% through the ammonium sulfate saturation ratio of twice fractionation precipitation in the said step (4).
Because general zymic cell walls is thicker, ALDH is influenced by outside extreme environment easily and loses activity, and therefore filtering out the little yeast cell breaking method of ALDH effect of vigor is the key of dealing with problems.
The broken yeast cell wall removes has multigelation method, chemosmosis method, polishing, sonioation method etc.; Wherein sonioation method is to utilize hyperacoustic cavitation effect, mechanical effect with the fragmentation of zymic cell walls; This method and data by MoM and MEI; The pair cell crushing effect is good, the characteristics little to the ALDH effect of vigor.
Key of the present invention is a ultrasonication snowy peak dry yeast cell.Ultrasonication is that a kind of hyperacoustic cavitation effect and mechanical effect of utilizing reaches the purpose that effective broken yeast cell discharges ALDH, cooperates cooling measure can more effectively preserve the vigor of ALDH.
The present invention compared with prior art has the following advantages and beneficial effect:
1. the yeast that uses among the present invention is the snowy peak dry yeast, is bread yeast, edible, and be prone to cultivate, the cycle is short, and is cheap, helps reducing production costs.
2. the present invention discharges ALDH with ultrasonic disruption snowy peak dry yeast, thereby has improved the cytoclasis effect, has effectively preserved the vigor of ALDH, has improved the yeast utilization ratio.
3. the quality of the ALDH that produces of the present invention can meet or exceed domestic like product; Yeast behind the extraction ALDH still can be used as him and uses as system feed, leach protein etc.; Both improved the zymic utility value, a kind of preparation method with ALDH of independent intellectual property right was provided again.
Embodiment
For better understanding the present invention, below in conjunction with embodiment the present invention is done detailed description further, but the scope that the present invention requires to protect is not limited to the scope that embodiment representes.
Embodiment 1
The first step is got a certain amount of snowy peak dry yeast and is added the entry mixing in the ratio of 8% (W/V), in 35 ℃ of water-baths, hatches 15min, makes bacteria suspension.Bacteria suspension in 4 ℃, 5000r/min, centrifugal 5min, is abandoned supernatant, and zero(ppm) water cleans 1 time, and recentrifuge is got the promptly wet yeast of deposition, and it is subsequent use to be stored in 4 ℃ of refrigerators.
Second goes on foot the yeast that will wet is resuspended to 0.05mol/L PBS (Na by mass volume ratio 20% (W/V)
2HPO
4-KH
2PO
4, pH7.5) in, ultrasonic disruption in ice-water bath, condition: treatment capacity 20mL, power 300W, broken time 3s, off time 1s, total time 15min.With broken liquid centrifugal 15min under 4 ℃, 5000r/min condition, collect supernatant, be the ALDH crude enzyme liquid, reach 0.46U/mg through the vigor that detects ALDH.
The 3rd step was that 35% ammonium sulfate precipitation room temperature leaves standstill 1h with above-mentioned ALDH crude enzyme liquid through saturation ratio, and the centrifugal 10min of 10000r/min collects supernatant; Through saturation ratio is that 70% ammonium sulfate precipitation room temperature leaves standstill 1h; The centrifugal 10min of 10000r/min, collecting precipitation adds 5mL phosphate buffered saline buffer (concentration is 0.05mol/L pH7.5); With resolution of precipitate, reach 1.03U/mg through the ratio vigor that detects ALDH.
The resolution of precipitate liquid of the 4th step with above-mentioned collection separates through Sephadex G-75; Elutriant be phosphate buffered saline buffer (concentration 0.05mol/L, pH7.5), flow velocity is 0.4mL/min; Collect elution peak; Ratio vigor through measuring ALDH reaches 1.24U/mg, through cataphoretic determination and standard substance the common band is arranged, and the ALDH molecular weight of measuring is about 57kDa.
Embodiment2
The first step is got a certain amount of snowy peak dry yeast and is added the entry mixing in the ratio of 10% (W/V), in 35 ℃ of water-baths, hatches 15min, makes bacteria suspension.Bacteria suspension in 4 ℃, 5000r/min, centrifugal 5min, is abandoned supernatant, and zero(ppm) water cleans 1 time, and recentrifuge is got the promptly wet yeast of deposition, and it is subsequent use to be stored in 4 ℃ of refrigerators.
Second goes on foot the yeast that will wet is resuspended to 0.05mol/L PBS (Na by mass volume ratio 20% (W/V)
2HPO
4-KH
2PO
4, pH8.0) in, ultrasonic disruption in ice-water bath, condition: treatment capacity 20mL, power 300W, broken time 3s, off time 1s, total time 15min.With broken liquid centrifugal 15min under 4 ℃, 5000r/min condition, collect supernatant, be the ALDH crude enzyme liquid, reach 0.48U/mg through the vigor that detects ALDH.
The 3rd step was that 35% ammonium sulfate precipitation room temperature leaves standstill 1h with above-mentioned ALDH crude enzyme liquid through saturation ratio, and the centrifugal 10min of 10000r/min collects supernatant; Through saturation ratio is that 70% ammonium sulfate precipitation room temperature leaves standstill 1h; The centrifugal 10min of 10000r/min, collecting precipitation adds 5mL phosphate buffered saline buffer (concentration is 0.05mol/L pH8.0); With resolution of precipitate, reach 1.07U/mg through the ratio vigor that detects ALDH.
The resolution of precipitate liquid of the 4th step with above-mentioned collection separates through Sephadex G-75; Elutriant be phosphate buffered saline buffer (concentration 0.05mol/L, pH8.0), flow velocity is 0.4mL/min; Collect elution peak; Ratio vigor through measuring ALDH reaches 1.27U/mg, through cataphoretic determination and standard substance the common band is arranged, and the ALDH molecular weight of measuring is about 56kDa.
Embodiment3
The first step is got a certain amount of snowy peak dry yeast and is added the entry mixing in the ratio of 12% (W/V), in 35 ℃ of water-baths, hatches 15min, makes bacteria suspension.Bacteria suspension in 4 ℃, 5000r/min, centrifugal 5min, is abandoned supernatant, and zero(ppm) water cleans 1 time, and recentrifuge is got the promptly wet yeast of deposition, and it is subsequent use to be stored in 4 ℃ of refrigerators.
Second goes on foot the yeast that will wet is resuspended to 0.05mol/L PBS (Na by mass volume ratio 20% (W/V)
2HPO
4-KH
2PO
4, pH8.5) in, ultrasonic disruption in ice-water bath, condition: treatment capacity 20mL, power 300W, broken time 3s, off time 1s, total time 15min.With broken liquid centrifugal 15min under 4 ℃, 5000r/min condition, collect supernatant, be the ALDH crude enzyme liquid, reach 0.50U/mg through the vigor that detects ALDH.
The 3rd step was that 35% ammonium sulfate precipitation room temperature leaves standstill 1h with above-mentioned ALDH crude enzyme liquid through saturation ratio, and the centrifugal 10min of 10000r/min collects supernatant; Through saturation ratio is that 70% ammonium sulfate precipitation room temperature leaves standstill 1h; The centrifugal 10min of 10000r/min, collecting precipitation adds 5mL phosphate buffered saline buffer (concentration is 0.05mol/L pH8.5); With resolution of precipitate, reach 1.09U/mg through the ratio vigor that detects ALDH.
The resolution of precipitate liquid of the 4th step with above-mentioned collection separates through Sephadex G-75; Elutriant be phosphate buffered saline buffer (concentration 0.05mol/L, pH8.5), flow velocity is 0.4mL/min; Collect elution peak; Ratio vigor through measuring ALDH reaches 1.30U/mg, through cataphoretic determination and standard substance the common band is arranged, and the ALDH molecular weight of measuring is about 57kDa.
Embodiment4
The first step is got a certain amount of snowy peak dry yeast and is added the entry mixing in the ratio of 14% (W/V), in 35 ℃ of water-baths, hatches 15min, makes bacteria suspension.Bacteria suspension in 4 ℃, 5000r/min, centrifugal 5min, is abandoned supernatant, and zero(ppm) water cleans 1 time, and recentrifuge is got the promptly wet yeast of deposition, and it is subsequent use to be stored in 4 ℃ of refrigerators.
Second goes on foot the yeast that will wet is resuspended to 0.05mol/L PBS (Na by mass volume ratio 20% (W/V)
2HPO
4-KH
2PO
4, pH7.5) in, ultrasonic disruption in ice-water bath, condition: treatment capacity 20mL, power 300W, broken time 3s, off time 1s, total time 15min.With broken liquid centrifugal 15min under 4 ℃, 5000r/min condition, collect supernatant, be the ALDH crude enzyme liquid, reach 0.45U/mg through the vigor that detects ALDH.
The 3rd step was that 35% ammonium sulfate precipitation room temperature leaves standstill 1h with above-mentioned ALDH crude enzyme liquid through saturation ratio, and the centrifugal 10min of 10000r/min collects supernatant; Through saturation ratio is that 70% ammonium sulfate precipitation room temperature leaves standstill 1h; The centrifugal 10min of 10000r/min, collecting precipitation adds 5mL phosphate buffered saline buffer (concentration is 0.05mol/L pH7.5); With resolution of precipitate, reach 1.03U/mg through the ratio vigor that detects ALDH.
The resolution of precipitate liquid of the 4th step with above-mentioned collection separates through Sephadex G-75; Elutriant be phosphate buffered saline buffer (concentration 0.05mol/L, pH7.5), flow velocity is 0.4mL/min; Collect elution peak; Ratio vigor through measuring ALDH reaches 1.22U/mg, through cataphoretic determination and standard substance the common band is arranged, and the ALDH molecular weight of measuring is about 56kDa.
The foregoing description is a preferred implementation of the present invention; But embodiment of the present invention is not restricted to the described embodiments; Other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify; All should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (10)
1. a preparation method who from the snowy peak dry yeast, separates acetaldehyde dehydrogenase is characterized in that, comprises the steps:
(1) the snowy peak dry yeast is added mixing in the entry, in 30 ℃ ~ 40 ℃ water-bath, hatch 10min ~ 20min, get deposition again behind the high speed centrifugation and promptly get the yeast that wets; Wherein the volume ratio of the weight of snowy peak dry yeast and water is: 5% ~ 15%;
(2) will wet that to add concentration be that 0.05mol/L, pH are mixed with bacteria suspension in 7.5 ~ 8.5 the phosphate buffered saline buffer to yeast, centrifugal behind this bacteria suspension ultrasonic disruption, collect supernatant, be crude enzyme liquid; The condition of its ultrasonic disruption is: treatment capacity be 15mL ~ 25mL, 1 ℃ ~ 8 ℃ of temperature, ultrasonic power be 250W ~ 350W, each ultrasonic time 1 ~ 4s, pitch time 1 ~ 8s, total ultrasonic time 10 ~ 20min; The volume ratio of wet zymic weight and phosphate buffered saline buffer is 15% ~ 25%;
(3) be that 30% ~ 60% ammonium sulfate adds in the crude enzyme liquid of above-mentioned gained with saturation ratio, centrifugal, get its supernatant; Be that 65% ~ 85% ammonium sulfate adds in this supernatant with saturation ratio again, centrifugal, get its deposition; And in this deposition, add phosphate buffered saline buffer; After treating that deposition is dissolved fully, separate, can obtain required acetaldehyde dehydrogenase enzyme.
2. preparation method according to claim 1 is characterized in that, the weight of snowy peak dry yeast and the volume ratio of water are 8% ~ 12% in the said step (1).
3. preparation method according to claim 1 is characterized in that, the bath temperature that the snowy peak dry yeast is hatched in the said step (1) is 33 ℃ ~ 37 ℃, and incubation time is 13min ~ 17min.
4. preparation method according to claim 1 is characterized in that, the volume ratio of wet zymic weight and phosphoric acid buffer is 18% ~ 22% in the said step (2).
5. preparation method according to claim 1 is characterized in that, the treatment capacity of ultrasonic disruption is 18mL ~ 22mL.
6. preparation method according to claim 1 is characterized in that, the pH value of said phosphoric acid buffer is 7.8 ~ 8.2.
7. preparation method according to claim 1 is characterized in that, the temperature of ultrasonic disruption is 2 ℃ ~ 6 ℃.
8. preparation method according to claim 1 is characterized in that, the power of ultrasonic disruption is 280W ~ 320W.
9. preparation method according to claim 1 is characterized in that, the time of each ultrasonic disruption is 1 ~ 3s, pitch time to be that 1 ~ 5s, total time are 12 ~ 18min.
10. preparation method according to claim 1 is characterized in that, crude enzyme liquid is respectively 35% ~ 55% and 70% ~ 80% through the ammonium sulfate saturation ratio of twice fractionation precipitation in the said step (4).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210005420XA CN102533684A (en) | 2012-01-10 | 2012-01-10 | Method for separating acetaldehyde dehydrogenase from Xuefeng dried yeasts |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210005420XA CN102533684A (en) | 2012-01-10 | 2012-01-10 | Method for separating acetaldehyde dehydrogenase from Xuefeng dried yeasts |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102533684A true CN102533684A (en) | 2012-07-04 |
Family
ID=46341718
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210005420XA Pending CN102533684A (en) | 2012-01-10 | 2012-01-10 | Method for separating acetaldehyde dehydrogenase from Xuefeng dried yeasts |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102533684A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114748566A (en) * | 2022-04-08 | 2022-07-15 | 恩众(山东)数字健康发展有限公司 | Composition for dispelling effects of alcohol and protecting liver and application thereof |
-
2012
- 2012-01-10 CN CN201210005420XA patent/CN102533684A/en active Pending
Non-Patent Citations (4)
| Title |
|---|
| 吴桂英等: "酿酒酵母细胞破碎释放乙醛脱氢酶的研究", 《食品工业科技》 * |
| 周百灵等: "酿酒酵母胞浆中乙醛脱氢酶的提取和研究", 《酿酒科技》 * |
| 张彦青等: "响应面法优化酿酒酵母乙醛脱氢酶和乙醇脱氢酶酶活检测方法", 《中国酿造》 * |
| 杨伟伟等: "法尔凯干酵母乙醛脱氢酶的提取和纯化研究", 《酿酒科技》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114748566A (en) * | 2022-04-08 | 2022-07-15 | 恩众(山东)数字健康发展有限公司 | Composition for dispelling effects of alcohol and protecting liver and application thereof |
| CN114748566B (en) * | 2022-04-08 | 2023-12-29 | 恩众(山东)数字健康发展有限公司 | Anti-alcohol liver-protecting composition and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CA2438233A1 (en) | Glucosamine and method of making glucosamine from microbial biomass | |
| KR100799517B1 (en) | Fermented Rice Bran Composition | |
| Jain et al. | A review on different modes and methods for yielding a pentose sugar: xylitol | |
| CN104529738B (en) | A kind of ubiquinone10extraction preparation method | |
| Rao et al. | Isolation and characterization of xylitol-producing yeasts from the gut of colleopteran insects | |
| CN101358173A (en) | Aspergillus niger ZJUT712 and application thereof in arctium fruit preparation by solid-state fermentation | |
| CN100436589C (en) | Process for preparation of gallic acid by co-culture | |
| CN101538586A (en) | Method for producing yeast powder being rich in beneficial metallic elements by utilizing beer waste yeast | |
| CN101307338A (en) | Method for preparing reducing coenzyme Q10 | |
| CN102533684A (en) | Method for separating acetaldehyde dehydrogenase from Xuefeng dried yeasts | |
| CN102250970A (en) | Method for synthesizing 1,3-dioxyacetone by glycerol fermentation | |
| WO2006013736A1 (en) | Mutant yeast, method of producing glutathione-rich yeast, culture thereof, fraction thereof, yeast extract and glutathione-containing foods and drinks | |
| CN102178937B (en) | Pharmaceutical composition for protecting liver and benefiting spleen and preparation method thereof | |
| CN103911402B (en) | A kind of method utilizing beet fermenting lactic acid | |
| CN104845961A (en) | Immobilization and application of 1,3-dihydroxy acetone producing recombinant genetic engineering bacteria | |
| CN102286565B (en) | Preparation method of theaflavin monomer | |
| CN109619594A (en) | A kind of spirulina instant powder and preparation method thereof | |
| CN102994591B (en) | Method for preparing galactooligosaccharide co-produced ethanol | |
| CN110760458B (en) | Complex function microbial inoculum for fermentation | |
| CN102776258A (en) | Fermentation method for coproduction of S-adenosyl-L-methionine and glutathione | |
| BR102016002700B1 (en) | Process for producing xylitol from hemicellulosic hydrolyzate of macaúba cake (acrocomia aculeata) and brewery co-products, and use | |
| CN106434786A (en) | Extracellular polysaccharide preparation method using fermentation of lactobacillus casei | |
| CN104366488A (en) | Method for producing winter jujube multienzyme complex probiotic through winter jujube pomace | |
| CN100567499C (en) | Use the method for producing coenzyme Q 10 by enzyme engineering method | |
| CN110760463A (en) | Composition containing probiotics as well as preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20120704 |