CN1063485C - Method for extracting plant alkaline protease - Google Patents
Method for extracting plant alkaline protease Download PDFInfo
- Publication number
- CN1063485C CN1063485C CN 91103165 CN91103165A CN1063485C CN 1063485 C CN1063485 C CN 1063485C CN 91103165 CN91103165 CN 91103165 CN 91103165 A CN91103165 A CN 91103165A CN 1063485 C CN1063485 C CN 1063485C
- Authority
- CN
- China
- Prior art keywords
- yucca
- enzyme
- blade
- proteolytic enzyme
- extracting method
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 238000000034 method Methods 0.000 title claims abstract description 16
- 108091005658 Basic proteases Proteins 0.000 title abstract 2
- 102000004190 Enzymes Human genes 0.000 claims abstract description 21
- 108090000790 Enzymes Proteins 0.000 claims abstract description 21
- 239000001653 FEMA 3120 Substances 0.000 claims abstract description 16
- 235000004552 Yucca aloifolia Nutrition 0.000 claims abstract description 16
- 235000012044 Yucca brevifolia Nutrition 0.000 claims abstract description 16
- 235000017049 Yucca glauca Nutrition 0.000 claims abstract description 16
- 108091005804 Peptidases Proteins 0.000 claims abstract description 15
- 241000196324 Embryophyta Species 0.000 claims abstract description 14
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims abstract description 11
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims abstract description 6
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims abstract description 6
- 235000011130 ammonium sulphate Nutrition 0.000 claims abstract description 6
- 241001532059 Yucca Species 0.000 claims description 18
- 102000035195 Peptidases Human genes 0.000 claims description 9
- 239000000047 product Substances 0.000 claims description 7
- 241000131505 Gloriosa Species 0.000 claims description 5
- 238000001556 precipitation Methods 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 239000002994 raw material Substances 0.000 claims description 4
- 239000006228 supernatant Substances 0.000 claims description 4
- 238000007601 warm air drying Methods 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 2
- 239000000126 substance Substances 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 9
- 239000004365 Protease Substances 0.000 abstract description 7
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 abstract description 6
- 238000002360 preparation method Methods 0.000 abstract description 3
- 238000005352 clarification Methods 0.000 abstract description 2
- 238000001035 drying Methods 0.000 abstract description 2
- 244000295923 Yucca aloifolia Species 0.000 abstract 1
- 239000003599 detergent Substances 0.000 abstract 1
- 235000015203 fruit juice Nutrition 0.000 abstract 1
- 238000005185 salting out Methods 0.000 abstract 1
- 235000019419 proteases Nutrition 0.000 description 5
- 235000013399 edible fruits Nutrition 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 241000234280 Liliaceae Species 0.000 description 2
- 239000005018 casein Substances 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 108010082495 Dietary Plant Proteins Proteins 0.000 description 1
- 108090000270 Ficain Proteins 0.000 description 1
- 238000012870 ammonium sulfate precipitation Methods 0.000 description 1
- 235000013405 beer Nutrition 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 230000035617 depilation Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 235000019836 ficin Nutrition 0.000 description 1
- POTUGHMKJGOKRI-UHFFFAOYSA-N ficin Chemical compound FI=CI=N POTUGHMKJGOKRI-UHFFFAOYSA-N 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 238000001155 isoelectric focusing Methods 0.000 description 1
- 239000010985 leather Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Landscapes
- Enzymes And Modification Thereof (AREA)
- Non-Alcoholic Beverages (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
A method for extracting alkaline protease from plant comprises directly drying Grating tissue of leaf surface to obtain crude enzyme product, or squeezing juice, centrifuging, and salting out with ammonium sulfate to obtain enzyme preparation. The method is simple and practical, and the prepared yucca protease is suitable for pH 7-12.5 and has the highest activity at the pH 12. The enzyme can be used in detergent, fruit juice clarification, etc.
Description
The present invention relates to a kind of method of from plant, extracting new proteolytic enzyme.
Zymin has been widely used on industrial production.It is abundant that plant protease generally has source resource, cheap characteristics.Thereby easily developed in a large number.Plant protease of selling on the market such as papoid, bromeline, ficin etc. just are widely used in that silk comes unstuck, aspect such as leather depilation, clarify beer, tenderization and medicine.
Proteolytic enzyme has different special nature, has determined they self application valency to plant.Commercially available plant protease all is the proteolytic enzyme of neutral slant acidity at present, and is more stable under the neutral slant acidity condition of pH, just devitalization easily under alkaline condition.The extracting method of known plants proteolytic enzyme is different different because of plant, is to extract the effusive juice of fruit surface again through preparation as caricin; Bromeline is to get fruit or cane to press juice, again through saltout, technology such as filtration, chromatography carry out separation and purification, obtain fruit enzyme and stem enzyme respectively.
The objective of the invention is to seek that the simple extraction process of a kind of usefulness is prepared, under the pH alkaline condition the stable new plant albumen enzyme of enzyme activity.
The scheme that the object of the invention realizes is that to adopt Liliaceae (LILIACEAE) Yucca (YUCCA) plant leaf be raw material, and blade is cleaned airing, scrapes and gets green palisade tissue, directly palisade tissue is ground into the enzyme raw product after with warm air drying.Or scrape by above-mentioned technology and to press leaf after getting palisade tissue, add the water of one times of juice amount, left standstill centrifugal removal suspended substance 10 minutes.Clear liquid adds ammonium sulfate by 8% weight ratio and leaves standstill centrifugal removal precipitation after about 4 hours.Supernatant liquor is supplied ammonium sulfate to 50% saturation ratio.Leave standstill centrifugal collecting precipitation after 12 hours, vacuumize and be lyophilized into zymin.
The present invention adopts yucca spp to make the enzyme extraction raw material, and its kind is filamentosa, golden-rimmed yucca (Golden side Yucca).Plain edge yucca (Bare leaf Yucca) and gloriosa, through survey above-mentioned 4 kind of plant enzymic activitys with the DHT-Casain method, gloriosa kind activity is the highest.In the blade of gloriosa kind, enzyme mainly is present in normal expansion, the growth normal healthy leaf, and that not open white tender leaf and withered Lao Ye contain the enzyme amount is few.
Extract the yucca plant protease of preparation with the inventive method.Adapt to the pH scope in PH7～12.5, enzymic activity is the highest during pH12.This enzyme proves two proteolytic enzyme components through polyacrylamide gel electrophoresis, and the thin layer isoelectric focusing electrophoresis confirms that these two component iso-electric points respectively are pH9.77 and 9.61.The yucca plant protease can be used for aspects such as washing composition, juice clarification.
The present invention directly scrapes and gets yucca plant leaf palisade tissue drying and make the enzyme raw product, method is simple and practical, broken through the complicated technology that vegetable-protein enzyme extraction is in the past adopted, made enzyme raw product is active high, still stable under the pH alkaline condition, can be directly used in fields such as washing powder production.
Embodiment 1:
Get 30.05 kilograms in the ripe healthy blade of gloriosa, scrape and get 12.6 kilograms of green palisade tissues.Scrape get thing and put-mesh is in the mesh bag of 1 * 1 (mm), is dried to moisture content＜7% o'clock in the warm air drying room under 55 ℃, get dry thing with ball mill pulverize 3.94 kilograms of powdery enzyme raw product, surveying activity with DHT-casein method is 15718 μ/grams.
Embodiment 2:
Get 15 kilograms in ripe healthy golden-rimmed yucca blade, scrape and get 6.1 kilograms of green palisade tissues, make 1.87 kilograms of powdery enzyme raw product by embodiment 1 method, activity is 8700 μ/grams.
Embodiment 3:
Get 17 kilograms in ripe healthy light leaf yucca blade, scrape and get 7.0 kilograms of green palisade tissues, make 2.03 kilograms of powdery enzyme raw product by embodiment 1 method, activity is 9500 μ/grams.
Embodiment 4:
Get 3.2 kilograms in ripe healthy yucca fiber crops blade, scrape and get 0.97 kilogram of green palisade tissue, 410 milliliters of extracting juices, getting supernatant water after the juice centrifugal (2000 rev/mins) doubles behind the deionized water, take by weighing the ammonium sulfate of juice amount 8%, slowly add and be stirred to dissolving in the juice, continue again to stir centrifugal after room temperature leaves standstill 4 hours (2000 rev/mins) 10 minutes.Get the ammonium sulfate precipitation that supernatant water is added to 50% saturation ratio again, leave standstill centrifugal after 12 hours (2000 rev/mins), the collecting precipitation thing gets enzyme and sticks with paste 0.082 kilogram.Enzyme is stuck with paste and is got 0.043 kilogram of zymin through vacuum lyophilization, and surveying activity with DHT-casein method is 51300 μ/grams.
Claims (3)
1. the extracting method of a plant alkalescence proteolytic enzyme.It is characterized in that adopting the yucca spp blade is raw material, and blade is cleaned airing, scrapes and gets green palisade tissue, and palisade tissue is ground into the enzyme raw product after with warm air drying; Or palisade tissue squeezed the juice, adding centrifugal removal suspended substance behind the water of one times of juice amount, clear liquid adds ammonium sulfate by 8% weight ratio and leaves standstill the centrifugal removal precipitation in back, and supernatant liquor is supplied ammonium sulfate to 50% saturation ratio, leaves standstill the back centrifugal collecting precipitation, vacuumizes into zymin.
2. according to the extracting method of the described plant alkalescence proteolytic enzyme of claim 1, it is characterized in that using fila-mentosa, golden-rimmed yucca (Golden side yucca), plain edge yucca, (Bare leaf yucca) and glo-riosa make raw material.
3. according to the extracting method of claim 1 or 2 described plant alkalescence proteolytic enzyme.It is characterized in that adopting normal expansion, growth normal healthy blade.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 91103165 CN1063485C (en) | 1991-05-09 | 1991-05-09 | Method for extracting plant alkaline protease |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 91103165 CN1063485C (en) | 1991-05-09 | 1991-05-09 | Method for extracting plant alkaline protease |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1066469A CN1066469A (en) | 1992-11-25 |
| CN1063485C true CN1063485C (en) | 2001-03-21 |
Family
ID=4905943
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 91103165 Expired - Fee Related CN1063485C (en) | 1991-05-09 | 1991-05-09 | Method for extracting plant alkaline protease |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1063485C (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101787360B (en) * | 2010-01-20 | 2012-01-04 | 云南万芳生物技术有限公司 | Tobacco processing active complex enzyme preparation for improving and enhancing tobacco quality and preparation method thereof |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101785578B (en) * | 2010-01-14 | 2014-07-30 | 云南万芳生物技术有限公司 | Enzyme preparation using glucose oxidase as assistant for tobacco processing and preparation and application method |
| CN103468780B (en) * | 2013-09-16 | 2015-07-29 | 张家港市金源生物化工有限公司 | A kind of aspartic protease is saltoutd the detection method of effect |
| CN115583710A (en) * | 2022-07-19 | 2023-01-10 | 江苏驰佳环保科技有限公司 | Chelating agent and preparation method thereof |
-
1991
- 1991-05-09 CN CN 91103165 patent/CN1063485C/en not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101787360B (en) * | 2010-01-20 | 2012-01-04 | 云南万芳生物技术有限公司 | Tobacco processing active complex enzyme preparation for improving and enhancing tobacco quality and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1066469A (en) | 1992-11-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JPS608773B2 (en) | How to remove bitterness from protein hydrolyzate | |
| SE9203744L (en) | PROCEDURES FOR PREPARING A PHARMACEUTICAL COMPOSITION SELECTIVELY LOWING THE LIPID LEVEL OF BLOOD | |
| CN1077409C (en) | Insects protein capsule or powder medicine and preparation process thereof | |
| US5503832A (en) | Method for extracting the soluble material from oil-bearing beans or seeds | |
| CN1063485C (en) | Method for extracting plant alkaline protease | |
| KR20010083037A (en) | A liquefied plant salt of preparation method and liquefied plant salt | |
| WO1991017671A1 (en) | Taste modifying composition and stabilization of taste modifying substance | |
| KR101855129B1 (en) | Pork wet aging method | |
| KR20200108616A (en) | Cosmetic composition containing caviar extracts and cosmetic manufacturing method using therof | |
| CN1045154C (en) | Process for preparing young wheat powder | |
| KR100871669B1 (en) | The method for manufacturing salicomia herbaceal fermented sap | |
| CN114711431A (en) | Method for efficiently extracting vitamin C from roxburgh rose | |
| CN1100792C (en) | Longan polysaccharide extracting process | |
| CN100352839C (en) | Method for preparing insect protein with blood-fat reducing function | |
| CN110251449A (en) | Using hiruto freeze-dried powder as the preparation method of the activity extract gel product of raw material | |
| DE1492959A1 (en) | Process for the extraction of compounds which are present in oil-rich plant tissues | |
| CN1049673C (en) | Method of extracting material from | |
| CN1161839A (en) | Technology for extracting effective component of matsutake | |
| CN1465569A (en) | Method for raising total flavone content in propolis | |
| US1195152A (en) | Alfalfa extract | |
| CN1040657C (en) | Method of extracting superoxide dismutase (SOD) from corn | |
| KR0169290B1 (en) | The essence of a chiness matrimony vine and its production | |
| CN1446483A (en) | Technique for processing medlar fine powder | |
| RU2296132C2 (en) | Method for polysaccharide production | |
| CN1331481C (en) | Method for extracting medicinal and edible substance from dictyoptera |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C15 | Extension of patent right duration from 15 to 20 years for appl. with date before 31.12.1992 and still valid on 11.12.2001 (patent law change 1993) | ||
| OR01 | Other related matters | ||
| C19 | Lapse of patent right due to non-payment of the annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |