CN100366737C - Production of curing lactase - Google Patents
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- CN100366737C CN100366737C CNB2006100097568A CN200610009756A CN100366737C CN 100366737 C CN100366737 C CN 100366737C CN B2006100097568 A CNB2006100097568 A CN B2006100097568A CN 200610009756 A CN200610009756 A CN 200610009756A CN 100366737 C CN100366737 C CN 100366737C
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- 108010005774 beta-Galactosidase Proteins 0.000 title claims abstract description 32
- 102100026189 Beta-galactosidase Human genes 0.000 title claims abstract description 30
- 108010059881 Lactase Proteins 0.000 title claims abstract description 29
- 229940116108 lactase Drugs 0.000 title claims abstract description 29
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 12
- 239000008101 lactose Substances 0.000 claims abstract description 28
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims abstract description 27
- 108090000790 Enzymes Proteins 0.000 claims abstract description 25
- 102000004190 Enzymes Human genes 0.000 claims abstract description 25
- 229940088598 enzyme Drugs 0.000 claims abstract description 25
- 239000007788 liquid Substances 0.000 claims abstract description 15
- 238000002360 preparation method Methods 0.000 claims abstract description 14
- 239000000725 suspension Substances 0.000 claims abstract description 9
- 235000014663 Kluyveromyces fragilis Nutrition 0.000 claims abstract description 6
- 241000235650 Kluyveromyces marxianus Species 0.000 claims abstract description 6
- 238000010257 thawing Methods 0.000 claims abstract description 6
- 239000000243 solution Substances 0.000 claims description 45
- 210000004027 cell Anatomy 0.000 claims description 23
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
- 241000894006 Bacteria Species 0.000 claims description 9
- UKVHQGHRJIEIML-UHFFFAOYSA-L calcium boric acid dichloride Chemical compound [Cl-].[Cl-].[Ca+2].OB(O)O UKVHQGHRJIEIML-UHFFFAOYSA-L 0.000 claims description 9
- 239000006285 cell suspension Substances 0.000 claims description 9
- 238000003756 stirring Methods 0.000 claims description 7
- 102000035195 Peptidases Human genes 0.000 claims description 6
- 108091005804 Peptidases Proteins 0.000 claims description 6
- 239000004372 Polyvinyl alcohol Substances 0.000 claims description 6
- 229920002451 polyvinyl alcohol Polymers 0.000 claims description 6
- 210000001082 somatic cell Anatomy 0.000 claims description 6
- 238000005303 weighing Methods 0.000 claims description 6
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 4
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims description 4
- 239000004327 boric acid Substances 0.000 claims description 4
- FYGDTMLNYKFZSV-URKRLVJHSA-N (2s,3r,4s,5s,6r)-2-[(2r,4r,5r,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,4r,5r,6s)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1[C@@H](CO)O[C@@H](OC2[C@H](O[C@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-URKRLVJHSA-N 0.000 claims description 3
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims description 3
- 229920002498 Beta-glucan Polymers 0.000 claims description 3
- 101710130006 Beta-glucanase Proteins 0.000 claims description 3
- 208000034189 Sclerosis Diseases 0.000 claims description 3
- 239000001110 calcium chloride Substances 0.000 claims description 3
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 3
- 230000008014 freezing Effects 0.000 claims description 3
- 238000007710 freezing Methods 0.000 claims description 3
- 238000000703 high-speed centrifugation Methods 0.000 claims description 3
- 238000007654 immersion Methods 0.000 claims description 3
- 238000012423 maintenance Methods 0.000 claims description 3
- 239000011259 mixed solution Substances 0.000 claims description 3
- 235000010413 sodium alginate Nutrition 0.000 claims description 3
- 239000000661 sodium alginate Substances 0.000 claims description 3
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- 235000013336 milk Nutrition 0.000 abstract description 18
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- 108010093096 Immobilized Enzymes Proteins 0.000 abstract description 5
- 238000000034 method Methods 0.000 description 12
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- 235000020247 cow milk Nutrition 0.000 description 6
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- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
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- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
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- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 210000000481 breast Anatomy 0.000 description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 2
- 201000006549 dyspepsia Diseases 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 210000000110 microvilli Anatomy 0.000 description 2
- 235000008476 powdered milk Nutrition 0.000 description 2
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- 206010000087 Abdominal pain upper Diseases 0.000 description 1
- 208000015943 Coeliac disease Diseases 0.000 description 1
- 108700016492 Congenital Lactase Deficiency Proteins 0.000 description 1
- 208000011231 Crohn disease Diseases 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 206010012742 Diarrhoea infectious Diseases 0.000 description 1
- 206010067715 Gastrointestinal sounds abnormal Diseases 0.000 description 1
- 206010064571 Gene mutation Diseases 0.000 description 1
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- 235000000824 malnutrition Nutrition 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000003094 microcapsule Substances 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 208000015380 nutritional deficiency disease Diseases 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 201000009868 osmotic diarrhea Diseases 0.000 description 1
- 208000028719 osmotic diarrheal disease Diseases 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
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- Dairy Products (AREA)
Abstract
The present invention relates to a production method of immobilized lactase, which comprises the following steps of preparation of Kluyveromyces fragilis suspension, enzyme wall prebreaking treatment, freeze thawing and wall breaking, final wall breaking of a high-pressure refiner, preparation of immobilized vector solution, preparation of lactase immobilized vector solution, preparation of immobile liquid, immobilized treatment, etc. The present invention uses immobilized lactase to produce low-lactose milk. The present invention is characterized in that continuous treatment can be carried out, and immobilized enzymes can not lose along with liquid milk and can be recovered to be repeatedly used so that the production cost of lactose milk is reduced.
Description
(1) technical field
What the present invention relates to is a kind of production method of microbial preparation, the production method of specifically a kind of microorganism cells broken wall and immobilized lactase.
(2) background technology
The enzyme that can reduce lactose among the human small intestine has Sumylact L (lactase) and beta-galactosidase enzymes, and (β-galactosidase), Sumylact L is positioned at the mucous membrane of small intestine brush border, and beta-galactosidase enzymes is positioned at lysosome, can not decompose the lactose in the enteric cavity.Lactase deficiencies is meant that the membrane bound enzyme of intestinal brush border chorionic cells is that lactase activity is low.Lactase deficiencies has three types: (1) congenital lactase deficiency: being meant the low or shortage of body lactase activity when being born, is on the body euchromosome due to the recessive gene, and this type is rarely found.(2) secondary lactase deficiency: be meant that the temporary lactase activity that causes the small intestine epithelium damage to cause owing to a variety of causes is low, as: diseases such as infectious diarrhea, immunoglobulin deficiency, celiac disease, regional ileitis, malnutrition, can after the organism disease rehabilitation, recover normal.(3) primary lactase deficiencies (claiming the adult type lactase deficiencies again): be modal a kind of, not by due to the influence of disease, mainly be to cause because lactase activity reduced gradually with the age, time of origin because of race with different from different places, behind weaning baby, just begin to take place, begin to occur during 20 years old left and right sides having.Population of China is many to be taken place in the time of 7～8 years old.It is generally acknowledged that lactase deficiencies is relevant with the different genetic gene mutations that cause of food habits that form from generation to generation.The lactase deficiencies incidence changes with country variant, different ethnic populations' variation.Europe crowd's 30% above lactase deficiencies, the asian population lactase deficiencies accounts for 75%～100%, the white American 12%, Black people 70%, Africa 90%～100%; Japan 100%, 3～13 years old children's lactase deficiencies incidence of China is 87%.
After lactose in the cow's milk entered small intestine, because the shortage of Sumylact L, lactose can not be broken down into glucose and semi-lactosi is absorbed, and is called lactose maldigestion and lactose malabsorption.After lactose enters colon, generated short chain organic acid such as acetic acid, propionic acid, butyric acid etc. and gas such as methane, H by fermentation using bacteria
2, CO
2Deng, the lactose fermentation process can cause borborygmus, stomachache, rectum gas and osmotic diarrhea, is called lactose intolerance when having these symptoms, serious lactose maldigestion or malabsorption are many to be taken place to a few hours after taking in a certain amount of lactose in 30 minutes.Lactose intolerance is bigger to infant's influence, semi-lactosi is the important substance of brain normal development, Sumylact L can not be decomposed into glucose with lactose and semi-lactosi is absorbed owing to lack, therefore, children's growth retardation, brain development also there is certain influence, and can be simultaneously with phenomenons such as vomitings, the adult is sometimes with the reaction of feeling sick.Have the people about 20% to have the lactose intolerance symptom among the lactase deficiencies person, and lactose intolerance symptom individual difference is very big.The lactose yield of what and severity of intolerance shape and the interior lactase activity of small intestine, absorption and whether to take in other based food simultaneously relevant, so produce low lactose milk and have crucial meaning.
But it is more expensive to produce the used Sumylact L price of low lactose milk, and this has just increased the production cost of liquid milk and milk powder, so low lactose milk is on the high side, and market sale is subjected to certain influence, if adopt immobilized lactase just can address this problem.Because immobilized lactase can not run off with milk liquid, can reclaim repeatedly and use, reduce production costs greatly.
(3) summary of the invention
The object of the present invention is to provide a kind of can the processing continuously, immobilized enzyme can not run off with liquid milk, can reclaim repeated use, reduces the production of curing lactase of the production cost of low lactose milk.
The object of the present invention is achieved like this:
1, the preparation of Kluyveromyces fragilis bacteria suspension:
To cultivate sophisticated Kluyveromyces fragilis cell through the 10000r/min high speed centrifugation, and obtain fresh active somatic cells, somatic cells with sterilized water washing 1-2 time, is mixed with the cell bacteria suspension with sterilized water then, cell concn is 5.0%-40.0%;
2, the pre-broken wall treatment of enzyme:
Get above-mentioned bacteria suspension 500mL, add proteolytic enzyme and beta-glucanase, proteolytic enzyme consumption 1.0-150.0u/g wet thallus, beta-glucan enzyme dosage 1.0-100u/g wet thallus, pH6.0-7.0,30 ℃-65 ℃ are incubated 1-6 hour;
3, freeze thawing broken wall
Cell suspension after enzyme is handled is taken out, places-20 ℃--quick freezing under 50 ℃ of temperature, it is thoroughly freezed, again the refrigerated cell is placed 40 ℃-75 ℃ to melt rapidly, obtain the frozen-thawed cell suspension;
4, the ultimate broken wall of high pressure homogenizer
Place liquid shear shredder assembly high pressure homogenizer to carry out the secondary broken wall cell suspension of freeze thawing, the temperature maintenance of ingress is at 20 ℃-45 ℃, the temperature in exit is not higher than 50 ℃, pressure maintains 30MPa-120MPa, obtain the active lactose enzyme solution of no cell structure, be called Ls solution;
5, preparation fixation support solution
Take by weighing 3.5g-20.0g polyvinyl alcohol (PVA), the 0.5g-10.0g sodium alginate adds 100mL distilled water, stirs 10 minutes, and static immersion 2-24 hour places under 121 ℃ of temperature and sterilized 20 minutes, is cooled to 10 ℃-35 ℃, and this solution is Ps solution;
6, preparation Sumylact L fixation support solution
Measure above-mentioned Ls solution and Ps solution, Ps: Ls is by 1: 1-1: 4, under aseptic condition, thoroughly stir, and this solution is called Ps-Ls solution;
7, preparation stationary liquid
Take by weighing boric acid 1.0-4.0g, calcium chloride 0.5-6.0g adds 100mL water, and heated and stirred is dissolved it, and then, 121 ℃ of temperature were sterilized 20 minutes, were cooled to 10 ℃-35 ℃, and this solution is boric acid-calcium chloride mixed solution;
8, immobilization
Ps-Ls solution is placed nodulizer; gently splash in boric acid-calcium chloride solution, the ratio of Ps-Ls solution and boric acid-calcium chloride solution is by 1: 1-1: 8, and the sclerosis through 2-8 hour; pull out and use the sterilized water washed twice, promptly make the immobilized lactase of solid globules shape.
9, other process for fixation
Immobilized lactase can be done glomeration, thread, column, strip, bulk, film like or the like with other process for fixation.
10, produce low lactose milk with immobilized lactase
Handle fresh cow milk or sterilization breast with immobilized lactase, the ratio of immobilized lactase and cow's milk is 10.0%-100%, the pH nature, and temperature 4-50 ℃, the reaction times is 0.5-5 hour, can produce low lactose liquid milk and low lactose powdered milk.
The present invention adopts immobilization technology, utilize chemistry or physical means with free microorganism cells or enzyme molecule, the area of space that is positioned to limit does not make cell or enzyme molecule leak, and substrate molecule can free in and out, and makes cell and enzyme keep active and repeatedly used a kind of basic fundamental.Immobilization technology comprises enzyme immobilization technology and immobilized cell technology, and process for fixation roughly can be divided into absorption method, covalent attachment method, crosslinking and entrapping method etc., wherein uses comparatively general with entrapping method.Entrapping method can be divided into gel embedding method and microcapsule method, exactly cell is wrapped in the small grid of gel or in the ultra-filtration membrane of semi-permeable membranes polymkeric substance, this method is simple to operate, and less to enzyme and cytoactive influence, the fixed cell carrier intensity of making is higher.Immobilized enzyme has the following advantages: 1. easily with immobilization enzyme-to-substrate, product separately; 2. can carry out batch reactions and dress post successive reaction in a long time; 3. can improve the stability of enzyme; 4. enzyme reaction process can in addition strict control; 5. do not have the residual of enzyme in the product solution, simplified extraction process; 6. be more suitable in multienzymatic reaction than water-soluble enzyme; 7. can increase the yield of product, improve the quality of product; 8. the service efficiency of enzyme improves, and cost reduces.But immobilized enzyme also has its weak point, can only be used for water soluble substrate, and is applicable to the small molecules substrate, and macromolecule substrate is not suitable for; Compare unsuitable multienzymatic reaction with intact cell; Intracellular enzyme must pass through separating of cell wall breaking and enzyme etc.
Produce low lactose milk with immobilized lactase, be characterized in and handle continuously that immobilized enzyme can not run off with liquid milk, can reclaim repeated use, reduce the production cost of low lactose milk.The present invention also relates to handle fresh cow milk, sterilization breast, produce low lactose liquid milk and low lactose powdered milk with immobilized lactase.
(4) embodiment
Get 5000mL and cultivate sophisticated Kluyveromyces fragilis cell fermentation liquid, through the 10000r/min high speed centrifugation, obtain fresh active somatic cells, somatic cells is washed 1-2 time with sterilized water, be mixed with the cell bacteria suspension with sterilized water then, cell concn is 5.0%-40.0%.
Get bacteria suspension 500mL, add proteolytic enzyme and beta-glucanase, proteolytic enzyme consumption 50.0u/g wet thallus, beta-glucan enzyme dosage 20u/g wet thallus, 6.5,42 ℃ of insulations of pH 1.5 hours.
Cell suspension after enzyme is handled is taken out, place quick freezing under-25 ℃ of temperature, it is thoroughly freezed, again the refrigerated cell is placed under 50 ℃ of temperature and melt rapidly, obtain the frozen-thawed cell suspension.
Cell suspension with freeze thawing places liquid shear shredder assembly high pressure homogenizer to carry out the secondary broken wall again, the temperature maintenance of ingress is about 20 ℃, and the temperature in exit is not higher than 50 ℃, and pressure maintains 70MPa, obtain the active lactose enzyme solution of no cell structure, be called Ls solution.
The preparation of immobilized lactase
Take by weighing 13.0g polyvinyl alcohol (PVA), the 5.0g sodium alginate adds 100mL distilled water, stirs 10 minutes, and static immersion 5 hours places under 121 ℃ of temperature and sterilized 20 minutes, is cooled to 35 ℃, and this solution is Ps solution.
Take by weighing boric acid 1.0-4.0g, calcium chloride 0.5-6.0g adds 100mL water, and heated and stirred is dissolved it, and then, 121 ℃ of temperature were sterilized 20 minutes, were cooled to 10 ℃-35 ℃, and this solution is boric acid-calcium chloride mixed solution.
Measure above-mentioned Ls solution and Ps solution, Ps: Ls thoroughly stirs under aseptic condition by 1: 2 mixed, and this solution is called Ps-Ls solution.
500mL places nodulizer with Ps-Ls solution, gently splashes in boric acid～calcium chloride solution, and constantly stirs, and prevents adhesion.The ratio of Ps-Ls solution and boric acid-calcium chloride solution was by 1: 4, and the sclerosis through 3 hours is pulled out and used the sterilized water washed twice, promptly makes the immobilized lactase of solid globules shape.If continuously fast with Ps: Ls solution injects boric acid-calcium chloride solution can obtain thread immobilized lactase.
Immobilized lactase is packed in the container, and then add fresh cow milk, the ratio of immobilized lactase and cow's milk is 50.0%, the pH nature, and 42 ℃ of temperature, the reaction times is 2 hours, can make low lactose liquid milk.
Claims (1)
1. production of curing lactase is characterized in that:
(1) preparation of Kluyveromyces fragilis bacteria suspension:
To cultivate sophisticated Kluyveromyces fragilis cell through the 10000r/min high speed centrifugation, and obtain fresh active somatic cells, somatic cells with sterilized water washing 1-2 time, is mixed with the cell bacteria suspension with sterilized water then, cell concn is 5.0%-40.0%;
(2) the pre-broken wall treatment of enzyme:
Get above-mentioned bacteria suspension 500mL, add proteolytic enzyme and beta-glucanase, proteolytic enzyme consumption 1.0-150.0u/g wet thallus, beta-glucan enzyme dosage 1.0-100u/g wet thallus, pH 6.0-7.0,30 ℃-65 ℃ are incubated 1-6 hour;
(3) freeze thawing broken wall
Cell suspension after enzyme is handled is taken out, places-20 ℃--quick freezing under 50 ℃ of temperature, it is thoroughly freezed, again the refrigerated cell is placed 40 ℃-75 ℃ to melt rapidly, obtain the frozen-thawed cell suspension;
(4) the ultimate broken wall of high pressure homogenizer
Place liquid shear shredder assembly high pressure homogenizer to carry out the secondary broken wall cell suspension of freeze thawing, the temperature maintenance of ingress is at 20 ℃-45 ℃, the temperature in exit is not higher than 50 ℃, pressure maintains 30MPa-120MPa, obtain the active lactose enzyme solution of no cell structure, be called Ls solution;
(5) preparation fixation support solution
Take by weighing 3.5g-20.0g polyvinyl alcohol (PVA), the 0.5g-10.0g sodium alginate adds 100mL distilled water, stirs 10 minutes, and static immersion 2-24 hour places under 121 ℃ of temperature and sterilized 20 minutes, is cooled to 10 ℃-35 ℃, and this solution is Ps solution;
(6) preparation Sumylact L fixation support solution
Measure above-mentioned Ls solution and Ps solution, Ps: Ls is by 1: 1-1: 4, under aseptic condition, thoroughly stir, and this solution is called Ps-Ls solution;
(7) preparation stationary liquid
Take by weighing boric acid 1.0-4.0g, calcium chloride 0.5-6.0g adds 100mL water, and heated and stirred is dissolved it, and then, 121 ℃ of temperature were sterilized 20 minutes, were cooled to 10 ℃-35 ℃, and this solution is boric acid-calcium chloride mixed solution;
(8) immobilization
Ps-Ls solution is placed shaper, gently splash in boric acid-calcium chloride solution, the ratio of Ps-Ls solution and boric acid-calcium chloride solution is by 1: 1-1: 8, and the sclerosis through 2-8 hour is pulled out and is used the sterilized water washed twice, promptly makes the solid immobilized lactase.
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| Application Number | Priority Date | Filing Date | Title |
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| CNB2006100097568A CN100366737C (en) | 2006-03-01 | 2006-03-01 | Production of curing lactase |
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| CNB2006100097568A CN100366737C (en) | 2006-03-01 | 2006-03-01 | Production of curing lactase |
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| CN100366737C true CN100366737C (en) | 2008-02-06 |
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| WO2009062132A2 (en) | 2007-11-09 | 2009-05-14 | California Institute Of Technology | Immunomodulating compounds and related compositions and methods |
| US20110251156A1 (en) | 2010-04-07 | 2011-10-13 | Yue Shen | Vehicle for delivering a compound to a mucous membrane and related compositions, methods and systems |
| JP6273200B2 (en) | 2011-07-12 | 2018-01-31 | ザ・ブリガーム・アンド・ウーメンズ・ホスピタル・インコーポレーテッド | Lipid-containing PSA compositions, methods of isolation and methods of use |
| JP2016521284A (en) | 2013-05-10 | 2016-07-21 | カリフォルニア インスティチュート オブ テクノロジー | Prevention and treatment of colorectal cancer with probiotics |
| CN103642785B (en) * | 2013-11-25 | 2015-08-12 | 四川农业大学 | A kind of immobilization 3-phenoxy benzoic acid degrading enzyme and preparation method thereof |
| WO2016201342A1 (en) | 2015-06-10 | 2016-12-15 | California Institute Of Technology | Sepsis treatment and related compositions methods and systems |
| EP3337321A4 (en) | 2015-08-19 | 2019-07-17 | President and Fellows of Harvard College | Lipidated psa compositions and methods |
| CN105969756A (en) * | 2016-05-26 | 2016-09-28 | 南京农业大学 | Immobilized Helianthus tuberosus L. fructan excision hydrolytic enzyme and preparation method thereof |
| WO2018014012A1 (en) | 2016-07-15 | 2018-01-18 | President And Fellows Of Harvard College | Glycolipid compositions and methods of use |
| CN108064936A (en) * | 2016-11-11 | 2018-05-25 | 内蒙古伊利实业集团股份有限公司 | A kind of preparation method for the liquid diary product for reducing Mei Lade products |
| CN107252108B (en) * | 2017-06-27 | 2021-01-29 | 福建农林大学 | Lactase microcapsule and preparation method thereof |
Citations (2)
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| CN1258452A (en) * | 1998-12-25 | 2000-07-05 | 浙江医科大学 | Method of producing low-lactose milk with solid thermophilic bacteria lactase |
| CN1724662A (en) * | 2005-07-06 | 2006-01-25 | 江南大学 | A kind of process for fixation of microbiological lactase and application thereof |
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| CN1258452A (en) * | 1998-12-25 | 2000-07-05 | 浙江医科大学 | Method of producing low-lactose milk with solid thermophilic bacteria lactase |
| CN1724662A (en) * | 2005-07-06 | 2006-01-25 | 江南大学 | A kind of process for fixation of microbiological lactase and application thereof |
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| CN1818061A (en) | 2006-08-16 |
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