CN106434789A - Method for extracting soluble dietary fiber in coffee peel - Google Patents
Method for extracting soluble dietary fiber in coffee peel Download PDFInfo
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- CN106434789A CN106434789A CN201610901260.5A CN201610901260A CN106434789A CN 106434789 A CN106434789 A CN 106434789A CN 201610901260 A CN201610901260 A CN 201610901260A CN 106434789 A CN106434789 A CN 106434789A
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- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
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Abstract
The invention belongs to the technical field of plant extraction, and particularly relates to a method for extracting soluble dietary fiber in coffee peel. The extraction method comprises the following steps of 1, taking dried coffee peel; performing first-time crushing until the granularity is 20 to 40 meshes; then, second-time crushing is performed until the granularity is 60 to 100 meshes; performing third-time crushing until the granularity is 100 to 150 meshes; obtaining dried coffee peel powder; 2, taking and mixing the obtained dried coffee peel powder, pancreatin, pectinase and cellulase; then, performing enzymolysis; performing centrifugation; collecting filtering liquid; performing concentration and depositing in alcohol to obtain the soluble dietary fiber. Compared with the prior art, the method provided by the invention has the advantages that the three-section type physical shearing with the crushing granularities being 20 to 40 meshes, 60 to 100 meshes and 100 to 150 meshes is combined with a composite enzyme process of the pectinase, pancreatin and cellulase; the extraction rate (p is lower than 0.01) of the soluble dietary fiber in the coffee peel is obviously improved; no chemical pollution exists; the safety is high; the method is suitable for large-scale production.
Description
Technical field
The present invention relates to technical field of plant extraction, extract water soluble dietary fiber from Coffee pulp particularly to one kind
Method.
Background technology
Dietary fiber has multiple physiologically actives, has obtained accreditation both domestic and external at present, such as liposuction, water absorption and swelling
Property, be conducive to the prevention to obesity, the absorption of poisonous and harmful substances can be reduced with the occurrence probability of colon cancer, have substantially simultaneously
Reduce Blood Cholesterol concentration effect, to prevention and improve heart disease, the high fat of Diet that coronary artery causes because of hardening
Matter mass formed by blood stasis has important effect.In addition to above-mentioned physiological function, the shortage of dietary fiber is also bent with appendicitis, vein blood vessel
, renal calculuss and vesical calculuses, duodenal ulcer, ulcerative colitiss, GERD, hemorrhoid and Deep venou pipe thrombosis shape
The sickness rate of disease such as one-tenth and occurring degree have much relations, take in high fiber food and can protect body invading from these diseases
Evil.Dietary fiber can be divided into insoluble dietary fiber (IDF) and water soluble dietary fiber (SDF) by its dissolubility.A lot of grinds
Study carefully report to have confirmed, water soluble dietary fiber, in Constipation, colon cancer, reduces blood cholesterol level, removes to human body
Harmful substance increases the aspects such as sugar tolerance that insoluble dietary fiber is incomparable.
Coffee pulp is the by-product of coffee preliminary working, and according to statistics, China's Coffee pulp produces more than 40 ten thousand tons per year within 2015, its
After will with 150,000 tons/year increase it is contemplated that being up to 1,000,000 tons about to the year two thousand twenty.There are some researches show, cellulose in Coffee pulp
Content is more than 60%, and the water soluble dietary fiber therefrom extracting has the functional characteristics such as good water suction, water holding, absorption.
However, the comprehensive utilization ratio of current China Coffee pulp is extremely low, except minority is returned in addition to field as fertilizer, most of accumulation abandons.
At present, prior art has a lot with regard to the extracting method of water soluble dietary fiber, such as enzyme process, alkali extraction method (chemistry
Method), chemistry-enzyme combine extraction method and physical-chemical process etc., but these all concentrate on bean dregs, Testa Tritici, skin of Semen Maydis, Semen Maydiss
The raw materials such as core, Testa oryzae, it has been investigated that, these methods to the extraction effect of water soluble dietary fiber in Coffee pulp and are paid no attention to
Think.Applicant once adopted the single factor test and response surface optimization cellulose enzyme process extraction work to water soluble dietary fiber in Coffee pulp
Skill is optimized, but optimum process condition disclosed in it is also only 9.72% to the extraction ratio of water soluble dietary fiber, effect
Unsatisfactory, referring to " response phase method optimizes Coffee pulp water soluble dietary fiber extraction process and Study on functional properties " literary composition
Record.
Content of the invention
In consideration of it, the present invention provides a kind of side that can significantly improve water soluble dietary fiber extraction ratio in Coffee pulp
Method.
In order to realize foregoing invention purpose, the present invention provides technical scheme below：
Step 1：Take coffee dry fruit skin to carry out pulverizing for the first time, carry out second pulverizing after being 20～40 mesh to granularity, extremely
Granularity is pulverized for carrying out third time after 60～100 mesh, is 100～150 mesh to granularity, obtains coffee dry fruit corium farinosum；
Step 2：Take gained coffee dry fruit corium farinosum to mix with pectase and carry out the first enzymolysis, take filtering residue after filtration, with pancreatin
Carry out second enzyme solution with cellulase mixing, centrifugation, collect filtrate, concentrate precipitate with ethanol, obtain water soluble dietary fiber.
Preferably, described first time pulverizes slightly beats powder for powder beater.
Preferably, described pulverizing for the second time is ground for wet milk.
Preferably, described third time is pulverized grinding for colloid machine.
Preferably, the condition of the first enzymolysis and the second enzymolysis is specially described in step 2：30 DEG C～50 DEG C, pH value is 4
1～3h is digested under the conditions of～7.
Preferably, in terms of g/U, the dry weight of the skin of coffee dry fruit described in step 2 and described pectase, described pancreatin and institute
The ratio stating the enzyme activity of cellulase is 1：(200-2000)：(200-2000)：(40-400).
Preferably, centrifugation step described in step 2 is specially room temperature, 5000r/min～10000r/min centrifugation 5～
10min.
Preferably, concentrate precipitate with ethanol described in step 2 being specially：It is 1 according to filtrate and ethanol volume ratio:2.5～6.5 add
Enter ethanol, carry out precipitate with ethanol, filter, collect precipitation.
Preferably, also including before described step 1 taking coffee fresh fruit to obtain Coffee pulp through preliminary working, then dehydrate
The step becoming the coffee dry fruit skin of water content≤12%.
Preferably, also including the making beating that adds water, under the conditions of 40 DEG C～55 DEG C before described first enzymolysis, described second enzymolysis
The step of standing 0.5h～2h.
Preferably, also include before described concentration precipitate with ethanol reducing moisture in filtrate using revolving instrument to water content being
The step of 40-70%.
Extracting method for water soluble dietary fiber in prior art extracts to water soluble dietary fiber in Coffee pulp
The relatively low defect of rate, it is 20～40 mesh that the present invention adopts grinding particle size, 60～100 mesh, and the three-stage physics of 100～150 mesh is cut
Cutting is combined with the combined-enzyme method of pancreatin, pectase and cellulase carries to the water soluble dietary fiber in Coffee pulp
Take, other extracting method compared to existing technology, the method significantly improves carrying of the water soluble dietary fiber in Coffee pulp
Take rate (p<0.01), and no chemical contamination, safe, suitable large-scale production.
Brief description
Fig. 1 is the flow chart of the method present invention extracting water soluble dietary fiber from Coffee pulp.
Specific embodiment
The invention discloses from Coffee pulp water soluble dietary fiber method, those skilled in the art can use for reference this
Civilian content, is suitably modified technological parameter and realizes.Specifically, all similar replacements and change are to art technology
It is it will be apparent that they are considered as including in the present invention for personnel.The method of the present invention and application have been passed through preferably
Embodiment is described, and related personnel substantially can be to side as herein described in without departing from present invention, spirit and scope
Method and application are modified or suitably change and combine, and to realize and to apply the technology of the present invention.
Agents useful for same, raw material Jun Keyou city in the extracting method of water soluble dietary fiber in the Coffee pulp that the present invention provides
Field is buied.
With reference to embodiment, the present invention is expanded on further：
Embodiment 1 extracts water soluble dietary fiber from Coffee pulp
Pretreatment：Take the Coffee pulp that coffee fresh fruit obtains through preliminary working, dehydrate becomes the coffee that water content is 10%
Dry fruit skin；
Three-stage physical shear：Take coffee dry fruit skin, carry out pulverizing for the first time using powder beater, be 20～40 mesh to granularity
After carry out second pulverizing, second is pulverized and is ground using wet milk, carries out third time and pulverize after being 60～100 mesh to granularity,
Third time is pulverized and is ground to granularity using colloid machine is 100～150 mesh, obtains coffee cherry corium farinosum；
Making beating：Take coffee cherry corium farinosum 10g add water 50ml making beating, 45 DEG C standing 1.5h；
Enzymolysis：Add 0.45g pectase (20000u/g) 45 DEG C of pH value to be enzymolysis 1h under the conditions of 5.5, filter, take filtering residue to add
Water 50ml pulls an oar, 40 DEG C of standing 0.5h；It is subsequently adding 0.45g cellulase (20000u/g) and 0.25g pancreatin (4000u/g),
40 DEG C, pH value is enzymolysis 2h under the conditions of 5.5；
Centrifugation：Enzymolysis solution is passed through centrifuge room temperature condition, 6500r/min is centrifuged 8min, abandon filtering residue, collect filtrate；
Revolving is dried：Reducing moisture to water content in filtrate using revolving instrument is 55%；
Concentrate precipitate with ethanol：It is 1 according to filtrate and ethanol volume ratio:4.5 addition ethanol, carry out precipitate with ethanol, filter, collect precipitation,
Obtain water soluble dietary fiber.
Embodiment 2 extracts water soluble dietary fiber from Coffee pulp
Pretreatment：Take the Coffee pulp that coffee fresh fruit obtains through preliminary working, dehydrate becomes the coffee that water content is 8% to do
Peel；
Three-stage physical shear：Take coffee dry fruit skin, carry out pulverizing for the first time using powder beater, be 20～40 mesh to granularity
After carry out second pulverizing, second is pulverized and is ground using wet milk, carries out third time and pulverize after being 60～100 mesh to granularity,
Third time is pulverized and is ground to granularity using colloid machine is 100～150 mesh, obtains coffee cherry corium farinosum；
Making beating：Take coffee cherry corium farinosum 6g add water 50ml making beating, 45 DEG C standing 1h；
Enzymolysis：Add 0.32g pectase (20000u/g), 45 DEG C, pH value is enzymolysis 1h under the conditions of 6, filters, takes filtering residue to add
Water 50ml pulls an oar, 45 DEG C of standing 0.5h；It is subsequently adding 0.2g cellulase (20000u/g) and 0.2g pancreatin (4000u/g), 45
DEG C, pH value is enzymolysis 1h under the conditions of 6；
Centrifugation：Enzymolysis solution is passed through centrifuge room temperature, 9500r/min is centrifuged 4min, abandon filtering residue, collect filtrate；
Revolving is dried：Reducing moisture to water content in filtrate using revolving instrument is 40%；
Concentrate precipitate with ethanol：It is 1 according to filtrate and ethanol volume ratio:4 addition ethanol, carry out precipitate with ethanol, filter, collect precipitation, obtain
Obtain water soluble dietary fiber.
Embodiment 3 extracts water soluble dietary fiber from Coffee pulp
Pretreatment：Take the Coffee pulp that coffee fresh fruit obtains through preliminary working, dehydrate becomes the coffee that water content is 12%
Dry fruit skin；
Three-stage physical shear：Take coffee dry fruit skin, carry out pulverizing for the first time using powder beater, be 20～40 mesh to granularity
After carry out second pulverizing, second is pulverized and is ground using wet milk, carries out third time and pulverize after being 60～100 mesh to granularity,
Third time is pulverized and is ground to granularity using colloid machine is 100～150 mesh, obtains coffee cherry corium farinosum；
Making beating：Take coffee cherry corium farinosum 8g add water 50ml making beating, 42 DEG C standing 1.5h；
Enzymolysis：Add 0.45g/g pectase (20000u/g), 42 DEG C, pH value is enzymolysis 1h under the conditions of 5.7, filters, takes filter
Slag add water 50ml making beating, 45 DEG C standing 0.5h；It is subsequently adding 0.45g cellulase (20000u/g) and 0.3g pancreatin (4000u/
G), 45 DEG C, pH value is enzymolysis 2h under the conditions of 5.7；
Centrifugation：Enzymolysis solution is passed through centrifuge room temperature condition, 5000r/min is centrifuged 10min, abandon filtering residue, collect filtrate；
Revolving is dried：Reducing moisture to water content in filtrate using revolving instrument is 50%；
Concentrate precipitate with ethanol：It is 1 according to filtrate and ethanol volume ratio:4.5 addition ethanol, carry out precipitate with ethanol, filter, collect precipitation,
Obtain water soluble dietary fiber.
Embodiment 4 extracts water soluble dietary fiber from Coffee pulp
Pretreatment：Take the Coffee pulp that coffee fresh fruit obtains through preliminary working, dehydrate becomes the coffee that water content is 9% to do
Peel；
Three-stage physical shear：Take coffee dry fruit skin, carry out pulverizing for the first time using powder beater, be 20～40 mesh to granularity
After carry out second pulverizing, second is pulverized and is ground using wet milk, carries out third time and pulverize after being 60～100 mesh to granularity,
Third time is pulverized and is ground to granularity using colloid machine is 100～150 mesh, obtains coffee cherry corium farinosum；
Making beating：Take the 50ml making beating that adds water in coffee cherry corium farinosum 7g, 48 DEG C of standing 1h；
Enzymolysis：Add 0.32g pectase (20000u/g), 48 DEG C, pH value is enzymolysis 1h under conditions of 5.4, filters, takes filter
Slag add water 50ml making beating, 46 DEG C standing 0.5h；It is subsequently adding 0.2g cellulase (20000u/g) and 0.2g pancreatin (4000u/
G), 46 DEG C, pH value is enzymolysis 1h under the conditions of 5.4；
Centrifugation：Enzymolysis solution is passed through centrifuge room temperature condition, 9500r/min is centrifuged 4min, abandon filtering residue, collect filtrate；
Revolving is dried：Reducing moisture to water content in filtrate using revolving instrument is 48%；
Concentrate precipitate with ethanol：It is 1 according to filtrate and ethanol volume ratio:4 addition ethanol, carry out precipitate with ethanol, filter, collect precipitation, obtain
Obtain water soluble dietary fiber.
The embodiment 5 relatively extraction ratio to water soluble dietary fiber in Coffee pulp for the different cut modes
Comparative example 1：On the basis of the extracting method that the embodiment of the present invention 1 provides, three-stage thing that embodiment 1 is recorded
Reason shearing is changed to one-part form physical shear, specifically, is ground using colloid machine, and grinding particle size is 100～150 mesh.Other extract
Step is identical with embodiment 1 with technological parameter.
Comparative example 2：On the basis of the extracting method that the embodiment of the present invention 1 provides, three-stage thing that embodiment 1 is adopted
Reason shearing is changed to two-part physical shear, first paragraph physical shear：Powder is slightly beaten using powder beater, grinding particle size is 20～40
Mesh；Second segment physical shear：Ground using colloid machine, grinding particle size is 100～150.Other extraction steps and technological parameter are same
Embodiment 1 is identical.
Comparative example 3：On the basis of the extracting method that the embodiment of the present invention 1 provides, three-stage thing that embodiment 1 is recorded
Reason shearing is changed to two-part physical shear, first paragraph physical shear：Ground using wet milk, grinding particle size is 60～100 mesh；The
Two-stage nitration physical shear：Ground using colloid machine, grinding particle size is 100～150 mesh.Other extraction steps and technological parameter are with enforcement
Example 1 is identical.
Table 1 relatively different physical shear modes are to water soluble dietary fiber extraction ratio in Coffee pulp
Note：Using the analysis of Duncan's multiple range test method, same row difference letter representation significance
Difference, (lowercase letter P<0.05, n=3；Capitalization represents P<0.01, n=3)
From the data of table 2, with comparative example 1-3 utilize one-part form (100～150 mesh) and two-part (20～40 mesh,
100～150 mesh；60～100 mesh, 100～150 mesh) by broken for coffee dry fruit corium farinosum be 100～150 mesh physical shear mode phase
The application using three-stage physical shear (20～40 mesh, 60～100 mesh, 100～150 mesh) by broken for coffee dry fruit corium farinosum is ratio
The coffee dry fruit corium farinosum of 100～150 mesh, the pole that combined with the combined-enzyme method of pancreatin, pectase and cellulase is significantly increased
Extraction ratio (the p of water soluble dietary fiber in Coffee pulp<0.01).
Using the extracting method that embodiment 2-4 is recorded, the carrying out of water soluble dietary fiber in Coffee pulp is extracted, result
Close with the extraction ratio of the water soluble dietary fiber of embodiment 1, no significant difference (P ＞ 0.05).
Embodiment 6 compares the extraction ratio to dietary fiber in Coffee pulp for the Different Extraction Method
Extracting the conventional method of water soluble dietary fiber in prior art has enzyme process, chemical method, chemical-enzymatic, physical chemistry
Method and physical chemistry enzyme process, concrete extraction process is as follows：
Enzyme process：Take the phosphate buffer 20mL that sample 1g adds 0.2mol/L pH 6.9, add 5ml compound enzyme (fiber
Plain enzyme, pectase and pancreatin) stand 2h at 37 DEG C.After cooling, mixed liquor uses sodium hydroxide to adjust pH to 10, mistake after standing 2h
Filter.Add filtrate 200mL ethanol, stand overnight, precipitation washing with alcohol, to neutral, is soluble dietary after lyophilization
Fiber.
Chemical method：Take sample 1g to add 2% acetic acid 20mL, be subsequently added 5% hydrogen peroxide 10mL, mixing liquid is at 60 DEG C
Heating 2h, after cooling, mixed liquor uses sodium hydroxide to adjust pH to 10, filters after standing 2h.Add filtrate 200mL ethanol, quiet
Put overnight, precipitation washing with alcohol, to neutral, is water soluble dietary fiber after lyophilization.
Chemical-enzymatic：Take sample 1g to add 2% acetic acid 20mL, be subsequently added 5% hydrogen peroxide 10mL, mixing liquid is at 60 DEG C
Lower heating 2h, after cooling mixed liquor use sodium hydroxide adjust pH to 6.9, add 5ml compound enzyme (cellulase, pectase and
Pancreatin) stand 2h at 37 DEG C.After cooling, mixed liquor uses sodium hydroxide to adjust pH to 10, filters after standing 2h.Add filtrate
200mL ethanol, stands overnight, and precipitation washing with alcohol, to neutral, is water soluble dietary fiber after lyophilization.
Physical-chemical process：Take sample 1g to add 2% acetic acid 20mL, be subsequently added 5% hydrogen peroxide 10mL, ultrasonic at room temperature
15min, mixing liquid heats 2h at 60 DEG C, and after cooling, mixed liquor uses sodium hydroxide to adjust pH to 10, filters after standing 2h.
Add filtrate 200mL ethanol, stand overnight, precipitation washing with alcohol, to neutral, is soluble dietary fine after lyophilization
Dimension.
Physical chemistry enzyme process：Take sample 1g to add 2% acetic acid 20mL, be subsequently added 5% hydrogen peroxide 10mL, surpass at normal temperatures
Sound 15min, mixing liquid heats 2h at 60 DEG C, and after cooling, mixed liquor uses sodium hydroxide to adjust pH to 6.9, adds 5ml multiple
2h is stood at 37 DEG C of synthase (cellulase, pectase and pancreatin).After cooling, mixed liquor uses sodium hydroxide to adjust pH to 10, quiet
Filter after putting 2h.Add filtrate 200mL ethanol, stand overnight, precipitation washing with alcohol, to neutral, as may be used after lyophilization
Soluble dietary fiber.
There is provided using above-mentioned enzyme process, chemical method, chemical-enzymatic, physical-chemical process, physical chemistry enzyme process and the present invention
Extracting method extracts to the water soluble dietary fiber in Coffee pulp, compares Different Extraction Method to meals in Coffee pulp
The extraction ratio of fiber, the results are shown in Table 2.
Table 2 compares the extraction ratio to dietary fiber in Coffee pulp for the Different Extraction Method
Note：Using the analysis of Duncan's multiple range test method, same row difference letter representation significance
Difference, (lowercase letter P<0.05, n=3；Capitalization represents P<0.01, n=3)
The content recorded from table 2, compared enzyme process disclosed in technology, chemical method, chemical-enzymatic, physical-chemical process,
Physical chemistry enzyme process, the extracting method pole of the water soluble dietary fiber that the present invention provides considerably improves to can in Coffee pulp
Extraction ratio (the P of soluble dietary fiber<0.01), and no chemical contamination, safe, suitable large-scale production.
The above is only the preferred embodiment of the present invention it is noted that ordinary skill people for the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of method of water soluble dietary fiber of extracting from Coffee pulp is it is characterised in that comprise the steps：
Step 1：Take coffee dry fruit skin to carry out pulverizing for the first time, carry out second pulverizing after being 20～40 mesh to granularity, to granularity
Pulverize for carrying out third time after 60～100 mesh, be 100～150 mesh to granularity, obtain coffee dry fruit corium farinosum；
Step 2：Take gained coffee dry fruit corium farinosum to mix with pectase and carry out the first enzymolysis, take filtering residue after filtration, with pancreatin and fibre
Dimension plain enzyme mixing carries out second enzyme solution, and centrifugation is collected filtrate, concentrated precipitate with ethanol, obtain water soluble dietary fiber.
2. method according to claim 1 slightly beats powder it is characterised in that described first time pulverizes for powder beater.
3. method according to claim 1 is it is characterised in that described pulverizing for the second time is ground for wet milk.
4. method according to claim 1 is it is characterised in that described third time is pulverized as the grinding of colloid machine.
5. the method according to any one of claim 1-4 it is characterised in that described in step 2 first enzymolysis and second enzyme
The condition of solution is specially：30 DEG C～50 DEG C, pH value is enzymolysis 1～3h under the conditions of 4～7.
6. method according to claim 1 is it is characterised in that in terms of g/U, the dry weight of described coffee dry fruit skin and described pancreas
The ratio of the enzyme activity of enzyme, described pectase and described cellulase is 1：(200-2000)：(200-2000)：(40-400).
7. method according to claim 1 is it is characterised in that centrifugation step described in step 2 is specially under room temperature condition
5000r/min～10000r/min is centrifuged 5～10min.
8. preparation method according to claim 1 is specially it is characterised in that concentrating precipitate with ethanol described in step 2：According to filter
Liquid and ethanol volume ratio are 1:2.5～6.5 addition ethanol, carry out precipitate with ethanol, filter, collect precipitation.
9. method according to claim 1 is it is characterised in that also include before described step 1 taking coffee fresh fruit through preliminary working
Obtain Coffee pulp, the step that then dehydrate becomes the coffee dry fruit skin of water content≤12%.
10. method according to claim 1 is it is characterised in that also include before described first enzymolysis, described second enzymolysis
Add water making beating, the step standing 0.5h～2h under the conditions of 40 DEG C～55 DEG C.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107904264A (en) * | 2017-12-28 | 2018-04-13 | 嘉必优生物技术（武汉）股份有限公司 | A kind of method and lycopene product that lycopene is prepared using saccharomyces cerevisiae |
| CN112544755A (en) * | 2020-12-11 | 2021-03-26 | 云南农业大学 | Instant coffee fruit tea powder and preparation method thereof |
| CN112753943A (en) * | 2021-01-27 | 2021-05-07 | 上海理工大学 | Energy bar with inflammatory bowel disease regulating function and manufacturing method thereof |
-
2016
- 2016-10-17 CN CN201610901260.5A patent/CN106434789A/en active Pending
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| 胡荣锁，等。: "响应面法优化咖啡果皮可溶性膳食纤维提取工艺和功能特性研究", 《热带农业科学》 * |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107904264A (en) * | 2017-12-28 | 2018-04-13 | 嘉必优生物技术（武汉）股份有限公司 | A kind of method and lycopene product that lycopene is prepared using saccharomyces cerevisiae |
| CN112544755A (en) * | 2020-12-11 | 2021-03-26 | 云南农业大学 | Instant coffee fruit tea powder and preparation method thereof |
| CN112753943A (en) * | 2021-01-27 | 2021-05-07 | 上海理工大学 | Energy bar with inflammatory bowel disease regulating function and manufacturing method thereof |
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Application publication date: 20170222 |