CN108070631A - A kind of method of aspergillus oryzae conversion iron rod yam production diosgenin - Google Patents
A kind of method of aspergillus oryzae conversion iron rod yam production diosgenin Download PDFInfo
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- CN108070631A CN108070631A CN201711482259.4A CN201711482259A CN108070631A CN 108070631 A CN108070631 A CN 108070631A CN 201711482259 A CN201711482259 A CN 201711482259A CN 108070631 A CN108070631 A CN 108070631A
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- aspergillus oryzae
- iron rod
- culture medium
- rod yam
- conversion
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P33/00—Preparation of steroids
- C12P33/20—Preparation of steroids containing heterocyclic rings
Abstract
The invention belongs to biological technical fields, disclose a kind of method of aspergillus oryzae conversion iron rod yam production diosgenin.Method is as follows：Aspergillus oryzae is inoculated into the culture medium added with iron rod yam powder and cultivates, medium pH 6, and aspergillus oryzae inoculum concentration is 6%, and fermentation temperature is 30 DEG C, fermentation time 6d.Under this fermentation condition, the conversion ratio that diosgenin is converted into iron rod yam powder reaches 71.57%.Its advantage is：Diosgenin yield is high, does not generate acidic organic wastewater pollutant, makes it environment friendly and pollution-free in process of production, environmentally friendly.
Description
Technical field
The invention belongs to biological technical field, more particularly to a kind of aspergillus oryzae conversion iron rod yam production diosgenin
Method.
Background technology
Diosgenin is the important original of synthesis treatment cardiovascular disease medicine, steroid hormone class drug and class drug of practising contraception
Material, application prospect is very extensive, and more and more researchs show that diosgenin has induction colon cancer and osteosarcoma cell
The multiple efficacies such as apoptosis, anti-hyperlipidemia, platelet aggregation-against, anti-AIDS, anti arteriosclerosis.Main use industrial at present
Acid-hydrolysis method produces diosgenin, and such method is first with strong acid hydrolysis Dioscin glycosidic bond, then is extracted with organic solvent
Sapogenin.And in traditional art production process, there are a large amount of acid waste waters to generate, seriously pollute environment.To this both at home and abroad
Through beginning attempt to explore diosgenin is prepared using microbial method.Microorganism conversion is to utilize microorganism exogenous compound
The chemical reaction of structural modification is carried out, with reaction condition is mild, program is simple, product purity is high and free from environmental pollution
Feature.
Also there are some patents using microorganism conversion production diosgenin at present, such as：A kind of yam saponin penicillium notatum
And preparation method thereof, it is a kind of using productivity of cultivation of cell dioscorea opposita sapogenin improve culture cell diosgenin yield method,
Turned using environment-friendly extraction process, Dioscorea nipponica Makino Fusarium sp and its application of Rhizopus oryzae extraction Dioscin, plant height effect
Change yellow ginger saponin(e bacterial strain and its application, it is a kind of by dioscorea zingiberensis wright-yellow ginger through microbial fermentation production Chinese yam saponin method, one
The peltate yam endophytic bacterium of kind production saponin(e enzyme and application, one plant of clostridium perfringen clx-74 bacterial strain and its use for producing diosgenin
On the way, living things catalysis prepares the method for diosgenin and its microbial inoculum used.On the special of microorganism conversion production diosgenin
It is sharp few, and there are some defects, the conversion ratio such as Rhizopus oryzae is low, and Determination of Diosgenin only has 2.79%, and indivedual patents are adopted
With joint sequential fermentation mode, method is complicated, and fermentation time is up to 46 days.
The content of the invention
In order to overcome above-mentioned shortcoming and deficiency of the prior art, primary and foremost purpose of the invention is to provide a kind of aspergillus oryzae
The method for converting iron rod yam production diosgenin, the method increase the conversion ratio of diosgenin.
The purpose of the present invention is realized by following proposal：
A kind of method of aspergillus oryzae conversion iron rod yam production diosgenin, including step in detail below：
S1. aspergillus oryzae conversion culture medium is prepared：Aspergillus oryzae conversion culture medium is grown using iron rod yam powder as aspergillus oryzae
Primary carbon source using tetracycline as bacteriostatic agent, adds vitamin, biotin, biological micromolecule substance, inorganic salts and minerals
Aspergillus oryzae to be supported to grow；
S2. aspergillus oryzae is screened：Including strain primary dcreening operation and strain secondary screening；Strain primary dcreening operation is that aspergillus oryzae is seeded to tablet primary dcreening operation
Culture medium, the single bacterium colony grown on picking tablet primary dcreening operation culture medium are rule again on tablet primary dcreening operation culture medium, third time tablet
Line is until isolate single bacterium colony；Strain secondary screening is that the single bacterium colony that strain primary dcreening operation is finally separating to obtain is inoculated into secondary screening culture medium
In cultivated；
S3. it is inoculated with, cultivates aspergillus oryzae：By aspergillus oryzae obtained by strain secondary screening in S2 be inoculated into the conversion culture medium in S1 into
Row fermented and cultured；
S4. the Aspergillus oryzae cell of diosgenin will be generated by centrifuging or filtering from culture by S3 fermented and cultureds
It is separated in base；
S5. add in petroleum ether after thalline separation to impregnate, petroleum ether phase is taken after centrifugation, is stored at room temperature crystallization, obtains aspergillus oryzae
Convert iron rod yam production diosgenin.
Aspergillus oryzae described in S1, which converts, contains 3.00g/L sodium nitrate in culture medium, 1.00g/L potassium dihydrogen phosphates,
0.50g/L epsom salts, 0.50g/L potassium chloride, 0.01g/L green-vitriols, 3.00g/L iron rod yam powder, 0.02g/L tetra-
Ring element, 0.1g/L vitamin Bs12, 5.5g/L glycine, 0.5g/L cellulases, 0.5g/L alpha-amylases.
The primary carbon source grown in aspergillus oryzae conversion culture medium using iron rod yam powder as aspergillus oryzae, wherein iron rod yam powder
In containing a variety of saponins substances for aspergillus oryzae conversion；
Sodium nitrate 3.00g/L, potassium dihydrogen phosphate 1.00g/L, seven water sulphur are added in tablet primary dcreening operation culture medium described in S2
Sour magnesium 0.50g/L, potassium chloride 0.50g/L, green-vitriol 0.01g/L, iron rod yam powder 3.00g/L, agar powder 20.00g/L.
Secondary screening culture medium described in S2 is fluid nutrient medium, containing sodium nitrate 3.00g/L, potassium dihydrogen phosphate 1.00g/L,
Epsom salt 0.50g/L, potassium chloride 0.50g/L, green-vitriol 0.01g/L, iron rod yam powder 3.00g/L.
The cultivation temperature of strain primary dcreening operation described in S2 is 28 DEG C~30 DEG C, incubation time 48h.
Strain secondary screening described in S2 is that one ring of single bacterium colony picking for being finally separating to obtain strain primary dcreening operation is sterile in 10mL
In physiological saline, with vortex mixer mixing, liquid-transfering gun pipettes 4mL bacterium solutions into secondary screening culture medium, with the rotating speed of 180r/min, 30 DEG C
Shaking table shaken cultivation 5 days.
The inoculum concentration of aspergillus oryzae described in S3 is 6%；The cultivation temperature of the fermented and cultured is 28 DEG C~35 DEG C, incubation time
It is 180r/min shaking table shaken cultivations in rotating speed for 6d.
The present invention is had the following advantages and advantageous effect compared with the prior art：
(1) present invention is further improved in the prior art, and the Chinese yam in iron rod yam is converted using aspergillus oryzae
Saponin(e produces diosgenin, optimizes working condition, to improve diosgenin yield and quality, solves to exist in traditional handicraft
Environmental pollution the problems such as a kind of new method is provided.
(2) present invention can modify Dioscin structure by mild mode, be converted into diosgenin, substitute sour water
Solution, does not generate waste water, can reduce energy consumption, reduce environmental pollution.
(3) present invention is with the addition of cellulase and amylase in the fermentation medium so that " package " is outside Dioscin
Starch and fiber be known as a degree of degradation so that Dioscin is more easy to dissolve out, aspergillus oryzae is facilitated to convert, improve potato
The conversion ratio of Chinese yam sapogenin；Using the method for the present invention, the conversion ratio that diosgenin is converted into iron rod yam powder reaches
71.57%.
(4) bacterial strain that the present invention uses can carry out large scale fermentation culture in a short time, easy to operate, and fermentation costs
Relatively low, the time is short, is limited from conditions such as time domain, seasons.
Specific embodiment
With reference to embodiment, the present invention is described in further detail, but the implementation of the present invention is not limited to this.
Sodium nitrate 3.00g/L, potassium dihydrogen phosphate 1.00g/ are added in the tablet primary dcreening operation culture medium used in following embodiment
L, epsom salt 0.50g/L, potassium chloride 0.50g/L, green-vitriol 0.01g/L, iron rod yam powder 3.00g/L, agar powder
20.00g/L。
The secondary screening culture medium used in following embodiment is fluid nutrient medium, contains sodium nitrate 3.00g/L, potassium dihydrogen phosphate
1.00g/L, epsom salt 0.50g/L, potassium chloride 0.50g/L, green-vitriol 0.01g/L, iron rod yam powder 3.00g/L.
Embodiment 1：
(1) aspergillus oryzae conversion culture medium is prepared：The primary carbon source that iron rod yam powder is grown as aspergillus oryzae is added in, content is
It is converted in 3.00g/L, wherein iron rod yam powder containing a variety of saponins substances for aspergillus oryzae.Tetracycline is added in culture medium to be prevented from contaminating
Upper miscellaneous bacteria, content 0.02g/L；The vitamin that aspergillus oryzae can be supported to grow, biotin, biological micromolecule object are added in culture medium
Matter, inorganic salts, minerals；Contain 3.00g/L sodium nitrate, 1.00g/L potassium dihydrogen phosphates, seven water sulphur of 0.50g/L in the culture medium
Sour magnesium, 0.50g/L potassium chloride, 0.01g/L green-vitriols, 3.00g/L iron rod yam powder, 0.02g/L tetracyclines, 0.1g/L dimensions
Raw element B12, 5.5g/L glycine, 0.5g/L cellulases, 0.5g/L alpha-amylases；
(2) aspergillus oryzae is seeded to tablet primary dcreening operation culture medium, 28 DEG C of biochemical cultivation case 4~7d of culture observe each bacterial strain life
Long result；After the different single bacterium colony picking of the form that will be grown on primary dcreening operation culture medium is rule on tablet again, third time
Plate streaking is until isolate single bacterium colony；
(3) it will be grown on primary dcreening operation plating medium or isolated one ring of single bacterium colony picking is sterile in 10ml
In physiological saline, with vortex mixer mixing, liquid-transfering gun pipettes 4ml bacterium solutions into fermentation medium, with the rotating speed of 180r/min, 30 DEG C
Shaking table shaken cultivation 5 days；
(4) aspergillus oryzae filtered out is inoculated into conversion culture medium and cultivated, inoculum concentration 6%, fermentation temperature is 30 DEG C.
Incubation time is 6d, is in rotating speed, 180r/min shaking table shaken cultivations.
(5) add in petroleum ether after thalline separation to impregnate, petroleum ether phase is taken after centrifugation, crystallization is stored at room temperature, to crystallized product
Carry out mass spectrum and nuclear magnetic resonance spectroscopy, the content of measurement diosgenin generation, if the conversion ratio of total soap can reach 71.57%.
HPLC measures its purity up to 96.5%.
Embodiment 2：
(1) aspergillus oryzae conversion culture medium is prepared.The primary carbon source that iron rod yam powder is grown as aspergillus oryzae is added in, content is
It is converted in 3.00g/L, wherein iron rod yam powder containing a variety of saponins substances for aspergillus oryzae.Tetracycline is added in culture medium to be prevented from contaminating
Upper miscellaneous bacteria, content 0.02g/L；The vitamin that aspergillus oryzae can be supported to grow, biotin, biological micromolecule object are added in culture medium
Matter, inorganic salts, minerals；Contain 3.00g/L sodium nitrate, 1.00g/L potassium dihydrogen phosphates, seven water sulphur of 0.50g/L in the culture medium
Sour magnesium, 0.50g/L potassium chloride, 0.01g/L green-vitriols, 3.00g/L iron rod yam powder, 0.02g/L tetracyclines, 0.1g/L dimensions
Raw element B12, 5.5g/L glycine, 0.5g/L cellulases, 0.5g/L alpha-amylases；
(2) aspergillus oryzae is seeded to tablet primary dcreening operation culture medium, 28 DEG C of biochemical cultivation case 4~7d of culture observe each bacterial strain life
Long result；After the different single bacterium colony picking of the form that will be grown on primary dcreening operation culture medium is rule on tablet again, third time
Plate streaking is until isolate single bacterium colony.
(3) it will be grown on primary dcreening operation plating medium or isolated one ring of single bacterium colony picking is sterile in 10ml
In physiological saline, with vortex mixer mixing, liquid-transfering gun pipettes 4ml bacterium solutions into fermentation medium, with the rotating speed of 180r/min, 30 DEG C
Shaking table shaken cultivation 5 days；
(4) aspergillus oryzae filtered out is inoculated into conversion culture medium and cultivated, inoculum concentration 6%, fermentation temperature is 32 DEG C.
Incubation time is 6d, is in rotating speed, 180r/min shaking table shaken cultivations.
(5) add in petroleum ether after thalline separation to impregnate, petroleum ether phase is taken after centrifugation, crystallization is stored at room temperature, to crystallized product
Carry out mass spectrum and nuclear magnetic resonance spectroscopy, the content of measurement diosgenin generation, if the conversion ratio of total soap can reach more than 70%.
Embodiment 3：
(1) aspergillus oryzae conversion culture medium is prepared.The primary carbon source that iron rod yam powder is grown as aspergillus oryzae is added in, content is
It is converted in 3.00g/L, wherein iron rod yam powder containing a variety of saponins substances for aspergillus oryzae.Tetracycline is added in culture medium to be prevented from contaminating
Upper miscellaneous bacteria, content 0.02g/L；The vitamin that aspergillus oryzae can be supported to grow, biotin, biological micromolecule object are added in culture medium
Matter, inorganic salts, minerals；Contain 3.00g/L sodium nitrate, 1.00g/L potassium dihydrogen phosphates, seven water sulphur of 0.50g/L in the culture medium
Sour magnesium, 0.50g/L potassium chloride, 0.01g/L green-vitriols, 3.00g/L iron rod yam powder, 0.02g/L tetracyclines, 0.1g/L dimensions
Raw element B12, 5.5g/L glycine, 0.5g/L cellulases, 0.5g/L alpha-amylases；
(2) aspergillus oryzae is seeded to tablet primary dcreening operation culture medium, 28 DEG C of biochemical cultivation case 4~7d of culture observe each bacterial strain life
Long result；After the different single bacterium colony picking of the form that will be grown on primary dcreening operation culture medium is rule on tablet again, third time
Plate streaking is until isolate single bacterium colony；
(3) it will be grown on primary dcreening operation plating medium or isolated one ring of single bacterium colony picking is sterile in 10ml
In physiological saline, with vortex mixer mixing, liquid-transfering gun pipettes 4ml bacterium solutions into fermentation medium, with the rotating speed of 180r/min, 30 DEG C
Shaking table shaken cultivation 5 days；
(4) aspergillus oryzae filtered out is inoculated into conversion culture medium and cultivated, inoculum concentration 6%, fermentation temperature is 35 DEG C.
Incubation time is 6d, is in rotating speed, 180r/min shaking table shaken cultivations.
(5) add in petroleum ether after thalline separation to impregnate, petroleum ether phase is taken after centrifugation, crystallization is stored at room temperature, to crystallized product
Carry out mass spectrum and nuclear magnetic resonance spectroscopy, the content of measurement diosgenin generation, if the conversion ratio of total soap can reach more than 70%.
Above-described embodiment is the preferable embodiment of the present invention, but embodiments of the present invention and from above-described embodiment
Limitation, other any Spirit Essences without departing from the present invention with made under principle change, modification, replacement, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (7)
- A kind of 1. method of aspergillus oryzae conversion iron rod yam production diosgenin, it is characterised in that including step in detail below：S1. aspergillus oryzae conversion culture medium is prepared：Aspergillus oryzae conversion culture medium is grown main using iron rod yam powder as aspergillus oryzae Carbon source, using tetracycline as bacteriostatic agent, add vitamin, biotin, biological micromolecule substance, inorganic salts and minerals to Support aspergillus oryzae growth；S2. aspergillus oryzae is screened：Including strain primary dcreening operation and strain secondary screening；Strain primary dcreening operation is that aspergillus oryzae is seeded to tablet primary dcreening operation culture Base, the single bacterium colony grown on picking tablet primary dcreening operation culture medium are rule again on tablet primary dcreening operation culture medium, third time plate streaking Until isolating single bacterium colony；Strain secondary screening be by the single bacterium colony that strain primary dcreening operation is finally separating to obtain be inoculated into secondary screening culture medium into Row culture；S3. it is inoculated with, cultivates aspergillus oryzae：It is sent out in the conversion culture medium that aspergillus oryzae obtained by strain secondary screening in S2 is inoculated into S1 Ferment culture；S4. the Aspergillus oryzae cell of diosgenin will be generated by centrifuging or filtering from culture medium by S3 fermented and cultureds It separates；S5. add in petroleum ether after thalline separation to impregnate, petroleum ether phase is taken after centrifugation, is stored at room temperature crystallization, obtain aspergillus oryzae conversion Iron rod yam produces diosgenin.
- 2. a kind of method of aspergillus oryzae conversion iron rod yam production diosgenin according to claim 1, it is characterised in that： Contain 3.00g/L sodium nitrate, 1.00g/L potassium dihydrogen phosphates, seven water sulphur of 0.50g/L in aspergillus oryzae conversion culture medium described in S1 Sour magnesium, 0.50g/L potassium chloride, 0.01g/L green-vitriols, 3.00g/L iron rod yam powder, 0.02g/L tetracyclines, 0.1g/L dimensions Raw element B12, 5.5g/L glycine, 0.5g/L cellulases, 0.5g/L alpha-amylases.
- 3. a kind of method of aspergillus oryzae conversion iron rod yam production diosgenin according to claim 1, it is characterised in that： Sodium nitrate 3.00g/L, potassium dihydrogen phosphate 1.00g/L, epsom salt are added in tablet primary dcreening operation culture medium described in S2 0.50g/L, potassium chloride 0.50g/L, green-vitriol 0.01g/L, iron rod yam powder 3.00g/L, agar powder 20.00g/L.
- 4. a kind of method of aspergillus oryzae conversion iron rod yam production diosgenin according to claim 1, it is characterised in that： Secondary screening culture medium described in S2 is fluid nutrient medium, contains sodium nitrate 3.00g/L, potassium dihydrogen phosphate 1.00g/L, seven water sulfuric acid Magnesium 0.50g/L, potassium chloride 0.50g/L, green-vitriol 0.01g/L, iron rod yam powder 3.00g/L.
- 5. a kind of method of aspergillus oryzae conversion iron rod yam production diosgenin according to claim 1, it is characterised in that： The cultivation temperature of strain primary dcreening operation described in S2 is 28 DEG C~30 DEG C, incubation time 48h.
- 6. a kind of method of aspergillus oryzae conversion iron rod yam production diosgenin according to claim 1, it is characterised in that： Strain secondary screening described in S2 is one ring of single bacterium colony picking for being finally separating to obtain strain primary dcreening operation in 10mL sterile salines In, with vortex mixer mixing, liquid-transfering gun pipettes 4mL bacterium solutions into secondary screening culture medium, with the rotating speed of 180r/min, 30 DEG C of shaking table vibrations Culture 5 days.
- 7. a kind of method of aspergillus oryzae conversion iron rod yam production diosgenin according to claim 1, it is characterised in that： The inoculum concentration of aspergillus oryzae described in S3 is 6%；The cultivation temperature of the fermented and cultured is 28 DEG C~35 DEG C, incubation time 6d, Rotating speed is 180r/min shaking table shaken cultivations.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109007691A (en) * | 2018-07-13 | 2018-12-18 | 河南京华食品科技开发有限公司 | Multistage enzymatic hydrolysis and spray drying technology combine the method for preparing the pure powder of iron rod yam |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101012474A (en) * | 2007-02-05 | 2007-08-08 | 大连理工大学 | Method for preparing Chinese yam saponin by microorganism transformation process |
| CN101698861A (en) * | 2009-07-29 | 2010-04-28 | 吉林农业大学 | Environment-friendly extraction process for extracting dioscin by adopting Rhizopus oryzae |
| CN103060416A (en) * | 2012-12-20 | 2013-04-24 | 华中科技大学 | Method for cleaning and producing dioscorea zingiberensis saponin with microbial technology adopted |
| CN103146795A (en) * | 2012-12-27 | 2013-06-12 | 西安绿泉生物技术有限公司 | Method for producing diosgenin through microbial fermentation of peltate yam rhizome-yellow ginger |
| CN106834407A (en) * | 2017-01-12 | 2017-06-13 | 华中科技大学 | A kind of method of bioanalysis green production turmeric saponin |
-
2017
- 2017-12-29 CN CN201711482259.4A patent/CN108070631A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101012474A (en) * | 2007-02-05 | 2007-08-08 | 大连理工大学 | Method for preparing Chinese yam saponin by microorganism transformation process |
| CN101698861A (en) * | 2009-07-29 | 2010-04-28 | 吉林农业大学 | Environment-friendly extraction process for extracting dioscin by adopting Rhizopus oryzae |
| CN103060416A (en) * | 2012-12-20 | 2013-04-24 | 华中科技大学 | Method for cleaning and producing dioscorea zingiberensis saponin with microbial technology adopted |
| CN103146795A (en) * | 2012-12-27 | 2013-06-12 | 西安绿泉生物技术有限公司 | Method for producing diosgenin through microbial fermentation of peltate yam rhizome-yellow ginger |
| CN106834407A (en) * | 2017-01-12 | 2017-06-13 | 华中科技大学 | A kind of method of bioanalysis green production turmeric saponin |
Non-Patent Citations (9)
| Title |
|---|
| TIANXIANG ZHENG: "Evaluation of different pretreatments on microbial transformation of saponins in Dioscorea zingiberensis for diosgenin production", 《BIOTECHNOLOGY & BIOTECHNOLOGICAL EQUIPMENT》 * |
| YU-LING ZHU: "Production of diosgenin from Dioscorea zingiberensis tubers through enzymatic saccharification and microbial transformation", 《APPL MICROBIOL BIOTECHNOL》 * |
| 曹友声: "《现代工业微生物学》", 31 March 1998 * |
| 李明军: "《提高山药商品性栽培技术问答》", 31 January 2013 * |
| 程永明: "《微生物实验》", 31 March 2015 * |
| 董悦生: "米曲霉直接转化盾叶薯蓣生产薯蓣皂苷元", 《过程工程学报》 * |
| 闫美屹: "生物法制备薯蓣皂素的清洁生产工艺", 《中国优秀硕士学位论文全文数据库（电子期刊）》 * |
| 雷攀: "生物酶预处理-醇提酸解法提取盾叶薯蓣中薯蓣皂苷元工艺研究", 《中国优秀硕士学位论文全文数据库（电子期刊）》 * |
| 齐珊珊: "米曲霉生物转化盾叶薯蓣中甾体皂苷的研究", 《中国优秀硕士学位论文全文数据库（电子期刊）》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109007691A (en) * | 2018-07-13 | 2018-12-18 | 河南京华食品科技开发有限公司 | Multistage enzymatic hydrolysis and spray drying technology combine the method for preparing the pure powder of iron rod yam |
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Application publication date: 20180525 |