alpha-Chymotrypsin
alpha-chymotrypsin from bovine pancreas
CAS: 9004-07-3
Molecular Formula: N/A
alpha-Chymotrypsin - Names and Identifiers
alpha-Chymotrypsin - Physico-chemical Properties
| Molecular Formula | N/A |
| Density | 1.37[at 20℃] |
| Melting Point | 127°C |
| Water Solubility | 125g/L at 25℃ |
| Solubility | Reconstitute in 1mM HCl. Soluble at 10mg/ml in 1mM HCl. 2mM calcium chloride serves as a stabilizer. Store aliquoted solutions at -20°C for up to a week. |
| Vapor Presure | 0Pa at 25℃ |
| Appearance | White to beige powder |
| Color | white |
| Merck | 13,2282 |
| Storage Condition | -20°C |
| MDL | MFCD00130481 |
| Physical and Chemical Properties | α-chymotrypsin is a proteolytic enzyme extracted from bovine or porcine pancreas. It is a white or white-like crystalline powder. No smell, easily soluble in water, insoluble in organic solvents. Isoelectric point PI = 8.1~8.6. It is rapidly inactivated in aqueous solutions. The Ph value of the 10% aqueous solution is about 3, and the aqueous solution of Ph3 ~ 4 is the most stable. The dry state is stable, and there is no decrease in vitality for 18 months under the condition of 40C. Chymotrypsin for injection is a sterile freeze-dried product of chymotrypsin, which is a white freeze-dried block. The aqueous solution is unstable and should be prepared freshly. |
| Use | For various surgical trauma after surgery, such as hematoma, edema, Hemarthrosis |
alpha-Chymotrypsin - Risk and Safety
| Risk Codes | R36/37/38 - Irritating to eyes, respiratory system and skin. R42/43 - May cause sensitization by inhalation and skin contact. R42 - May cause sensitization by inhalation |
| Safety Description | S26 - In case of contact with eyes, rinse immediately with plenty of water and seek medical advice. S36 - Wear suitable protective clothing. S36/37 - Wear suitable protective clothing and gloves. S24 - Avoid contact with skin. S22 - Do not breathe dust. |
| WGK Germany | 3 |
| RTECS | GC3050000 |
| FLUKA BRAND F CODES | 3-10 |
| TSCA | Yes |
| HS Code | 35079090 |
alpha-Chymotrypsin - Nature
Open Data Verified Data
- a proteolytic enzyme extracted from bovine or porcine pancreas, white or off-white crystalline powder. No smell, soluble in water, water solution will be rapidly inactivated. The pH value of 10% aqueous solution is about 3, and the aqueous solution of pH3 ~ 4 is the most stable. Dry stable, 4 deg C, 18 months can not reduce the vitality of the phenomenon.
- a-chymotrypsin isolated from bovine pancreas consists of 245 amino acid residues and has an isoelectric point pl = 8.1 to 8.6. The molecular weight of 42000, the optimum pH is 8~9. d a chymotrypsinogen is not activated by enterokinase, pepsin, calcium chloride and chymotrypsin itself can not make it activated. However, trypsin can activate prochymotrypsin into a-chymotrypsin, that is, five pairs of disulfide bonds in the molecule, a single peptide chain, which loses two 2 peptides after activation (silk 14. Jing 1 5 and Su 147. Asparagine 148) forms d-chymotrypsin consisting of three peptide chains. The active center amino acid residue is Group 37. Day 102. Silk 195, which acts on the protein to preferentially hydrolyze the peptide bond formed by the carboxyl groups of L-tyrosine and L-phenylalanine. It is an endopeptidase.
Last Update：2024-01-02 23:10:35
alpha-Chymotrypsin - Preparation Method
Open Data Verified Data
- Fresh Bovine pancreas was taken, minced, and extracted with 0.125 mol/L cold sulfuric acid solution with sufficient stirring. Ammonium sulfate was added to the extract to reach 40% saturation, the cold chamber was left overnight, the supernatant was siphered, the bottom layer was filtered with an appropriate amount of diatomite, and the supernatant and the filtrate were combined. Then, ammonium sulfate was further added so as to reach 70% of saturation, the cold chamber was left overnight, the supernatant was discarded, and the bottom precipitate was filtered. It was then dissolved in 3 times the amount of ice water, and the fractional precipitation with ammonium sulfate 40% and 70% saturation was repeated. The second 70% saturation precipitate cake was dissolved in 1.5-fold ice water, and 0.5-fold saturated ammonium sulfate solution was added thereto, followed by crystallization at 25 ° C. Adjusted to pH 5 with sodium hydroxide. After separation of the crystals, the crystals were dissolved in ice-cold water, adjusted to pH 2, and filtered. The filtrate was added with 2 times the amount of saturated ammonium sulfate solution, adjusted to pH 5, crystallized at room temperature, and filtered to obtain chymotrypsinogen crystals. The chymotrypsinogen was crystallized, and 3 times of ice-cold distilled water and sulfuric acid solution were added dropwise. After dissolution, pH 7.6 phosphate buffer and a certain amount of sodium hydroxide were added to maintain pH 7.6, then 5mg of trypsin was added to each log of chymotrypsin pro and activated at 5 °c for 48h. Then adjust the pH to 4 with sulfuric acid, add an appropriate amount of ammonium sulfate salting out, filtration, precipitation with 0. 005mol/L sulfuric acid dissolved at room temperature, static, there are a lot of crystal formation, filtration, can be obtained by chymotrypsin.
- see trypsin for porcine pancreas preparation method.
Last Update：2022-01-01 11:10:52
alpha-Chymotrypsin - Use
Open Data Verified Data
- proteolytic enzyme. For general wound or postoperative wound healing, anti-inflammatory and prevent local edema, blood, sprain hematoma, breast surgery, local swelling, otitis media, rhinitis; For cataract extraction, to reduce capsular rupture and retinal damage.
- usage and dosage of cataract extraction: with 5000/1 saline solution, the pupil is injected into the posterior chamber, after 2~3 minutes, after the Crystal is floating, rinse with saline, the lens can be removed. Intramuscular injection, 5mg each time, 1~2 times a day. Wet mist or spray: 5mg each time; Gas solution therapy, 0.2~0.3mg each time (with distilled water 2 ~ 3mL), 3~4 times a day; Tracheal instillation, with each ml containing this product, 0. 5mg solution, appropriate amount of drops.
Last Update：2022-01-01 11:10:52
alpha-Chymotrypsin - Safety
Open Data Verified Data
- intramuscular injection can cause anaphylactic shock, but also can cause temporary glaucoma, moderate uveitis and corneal edema. Injection site can produce pain and swelling, severe liver patients, coagulation function is not normal, less than 20 years old or vitreous body is not fixed in patients with traumatic cataract. No intravenous injection was allowed. The aqueous solution is unstable and should be freshly prepared. Allergy test should be done before use.
- light-shielding, closed, stored in a cool place.
Last Update：2022-01-01 11:10:53
alpha-Chymotrypsin - Trait
White or nearly white crystalline or amorphous powder. No odor. Soluble in water and dilute buffer solution.
Last Update：2023-08-16 21:32:38
alpha-Chymotrypsin - Vitality definition
The amount of enzyme that reduces the optical density value at 237nm by 0.001 per minute is 1Atee unit (u) at 25 °c, pH 7.0.
Last Update：2023-08-16 21:32:38
alpha-Chymotrypsin - Optimum pH
7.5～8.5。
Last Update：2023-08-16 21:32:38
alpha-Chymotrypsin - Reference Information
| LogP | -1.3 at 20℃ |
| EPA chemical substance information | information provided by: ofmpeb.epa.gov (external link) |
| Use | α-chymotrypsin can be used as a proteolytic enzyme. Generally used for wound or wound healing after surgery, anti-inflammatory and prevent local blood, edema, sprain hematoma, breast postoperative swelling, rhinitis, otitis media and so on. For cataract extraction, release of ciliary muscle ligaments, to reduce membrane rupture and retinal damage. However, allergy test must be done before use, not intravenous injection, injection site can be pain and swelling, intramuscular injection can cause anaphylactic shock, but also can cause moderate uveitis, corneal edema and transient glaucoma. Patients with severe liver, abnormal coagulation function, vitreous body is not fixed in patients with traumatic cataract and patients under the age of 20. Cataract extraction; With 1/5000 saline solution to rinse, the pupil is injected into the posterior chamber, after 2~3min, after the Crystal floating, with saline irrigation, you can take out the lens. Intramuscular injection, 5mg each time, 1~2 times a day, wet mist or spray: 5mg daily, gas solution therapy, 0.2~0.3mg each time (with distilled water 2 ~ 3ml), 3~3 days a day ~ 4 times; Tracheal instillation, with each ml containing 0.5mg solution of this product, the appropriate amount of drops. for biochemical research, protein structure analysis, decomposition and digestion, clinical for wound healing after trauma or surgery, anti-inflammatory and prevent local edema, blood, sprain, hematoma, otitis media, rhinitis, etc. The product is protein decomposition enzyme drugs. Can promote the elimination of blood clots, purulent secretions and ring dead tissue such as liquefaction, for eye surgery to release the ciliary ligament, reduce traumatic iridocyclitis. It can also be used for wound or local inflammation to reduce local secretion and edema. used for various surgical trauma such as hematoma, edema, hemarthrosis A serine protease that hydrolyzes peptide bonds with an aromatic or long hydrophobic chain (Tyr, Trp, phe, Met) peptide bond. Note: For proteolysis in the absence of exogenous trypsin activity. |
| production method | the pancreas was taken out within 1h after slaughter and leached with sulfuric acid at 0 °c. The leaching solution was graded salting out with ammonium sulfate, and the salting-out material was dissolved in cold distilled water (pH was adjusted with sulfuric acid), and then the pH was adjusted with ammonium sulfate and sodium hydroxide to make the crystallization complete, repeatedly dissolved and recrystallized to obtain chymotrypsin crystals. Chymotrypsin was obtained by crystallization activation, salting out, dialysis and drying. with Bovine pancreas as raw material, after extraction and salting-out, chymotrypsinogen was first prepared, activated by trypsin, and then purified by recrystallization with ammonium sulfate and ethanol. after crushing and extracting the cattle for slaughter, quickly remove the bovine pancreas (usually within 0.5~1H), wash and remove the fat, connective tissue and other debris, immediately immersed in pre-frozen sulfuric acid solution (0.125mol/L), suddenly cooled, stored at about 0C, to fill up 100kg after feeding. Take the pancreas and put it into a meat grinder to mince it into pancreatic plasma (if necessary, it should be twisted 3 times). Add 2 times the amount of ice-cold sulfuric acid (0.125mol/L). In the cold room, stir 1 times every 1~2H, soak and extract 24h. Filter the impregnated material with a coarse filter bag (or two layers of gauze). After the filter residue is dried, repeat the process with 1 times the amount of cold sulfuric acid solution (0.125mol/L), then filter and discard the filter residue, combine the filtrates twice, add solid ammonium sulfate to make the concentration of 242g per liter of filtrate, at which point the concentration of ammonium sulfate reaches 40% saturation, place in cold room overnight, siphon the supernatant, and add appropriate amount of diatomite as filter aid to the bottom precipitate, filtration was performed under reduced pressure, and the supernatant and filtrate were combined to obtain an extract. Bovine pancreas ice cold H2SO4 → extract salting-out and crystallization by grade, solid ammonium sulfate was added to every liter of extract to make the concentration of ammonium sulfate reach 70% saturation. The cold room was left overnight, add 3 times the weight of the cake ice water to dissolve, repeat with ammonium sulfate 40% and 70% saturation Fractional precipitation, take the second 70% saturation precipitation cake, add 1.5 times the weight of the cake ice water solution, and add 0.5 times the weight of the filter cake saturated ammonium sulfate solution, Adjust pH 5 with sodium hydroxide (5mol/L), stand in a 250C incubator for 48h for crystallization (take a drop of the crystallization solution and set it on a glass slide for observation with a microscope of 100 times, which should be obviously needle-like crystals), then the crystallization was filtered to dryness under reduced pressure to obtain crude chymotrypsin. The yield was 5% ~ 6% calculated according to the mass of bovine pancreas. Take the crude trypsinogen and add 7 times the amount of ice-cold distilled water to dissolve it, and add sulfuric acid (2.5mol/L) Dropwise to adjust the Ph to about 2. Set the solution in Buchner funnel and filter it with acid-washed talc, the filtrate should be clarified, and then add 2 times the amount of saturated ammonium sulfate solution, Dropwise adding sodium hydroxide (5mol/L) to neutralize the acidity so that the Ph is about 5, under the condition of 20 ~ 250C, holding for more than 4H, there is white precipitation, which is observed as rod-like crystals under the microscope, filtering and discarding the filtrate, and repeating the crystallization for 3 times according to this method, Chymotrypsinogen crystals were obtained. Extract (NH4)2SO4 → Fractional salting-out precipitation (NH4)2SO4;NaOH;Ph4 ^ → Crystal chymotrypsinogen activation, salting-out, crystal weighing chymotrypsinogen Crystal, 3 times this amount of ice-cold distilled water was added and sulfuric acid solution was added dropwise to dissolve, and then an equal amount of phosphate buffer solution (0.5mol/L) with pH 7.6 was added. And a certain amount of sodium hydroxide [equivalent to the mass of 2.5mol/L sulfuric acid in (2)] to maintain the Ph at 7.6, more than 150-fold trypsin 5mg per g of prochymotrypsin was added. It was placed in a refrigerator at 50C and allowed to activate for 48h. Adjust the Ph to about 4 with sulfuric acid (0.5mol/L), add 0.5 of solid ammonium sulfate per liter of activated solution, salt out and place for 2H, filter the solution under reduced pressure in Buchner funnel, and discard the filtrate, the precipitate was clarified with 3/4 times the amount of sulfuric acid (0.005mol/L), washed with talc powder, and a small amount of seed crystal was added ~ 250C standing for 24h, when a large number of crystals were generated, the chymotrypsin crystals were obtained by filtration. Chymotrypsinogen ice-cold; Trypsin; 50 → activated H2SO4;Ph4 → Salting out; Crystal chymotrypsin Crystal dialysis, sterilization, dry weighing chymotrypsin Crystal, adding 3 times the amount of distilled water, add sulfuric acid solution (0.005mol/L) Dropwise and dissolve it into a dialysis bag every 350ml, so that the dialysis bag is immersed in 50 water bath, and the inner and outer solutions are on the same plane, continuous dialysis with water 2 ~ 3d, dialysis should be complete. Chymotrypsin crystallization 50 → dialysis dialysate freezing; Decompression → Sterilization; Dried product |
| toxic substance data | information provided by: pubchem.ncbi.nlm.nih.gov (external link) |
Last Update：2024-04-09 21:54:55
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