本产品来源于表达经过定点突变优化的林伯氏白色念球菌（Engyodontium album）蛋白酶K基因的毕赤酵母，具有比活性高、产量高及更广的温度/pH值活性范围等优势。先进的蛋白质工程技术及28,000升酵母平台有效降低了单位成本，为生物医药研发与生产提供较高性价比的蛋白酶K。酵母高效表达和色谱纯化保证了产品的纯度，本产品不含细菌内毒素，应用范围比天然蛋白酶K更广泛。
蛋白酶K是现有蛋白酶中活性较高的品种，pH活性范围为4-12，温度范围达0-75℃，甚至在SDS、尿素或EDTA存在的情况下依旧保持活性。其可切割脂肪族氨基酸和芳香族氨基酸的羧基端肽键，能够降解所有蛋白质。
| 货号 | P1520269
|
| 产品名称 | 蛋白酶K溶液 (40 mg/mL)
|
来源种属
| 表达经过基因工程改造的白色念珠菌（Engyodontium album）蛋白酶K基因的毕赤酵母
|
分子量
| 29.3KD
|
等电点（PI）
| 8.9
|
稀释缓冲液
| 20mM Tris-HCl（pH 7.4），1mM CaCl₂；或20mM Tris-HCl（pH 7.4），1mM CaCl₂，2%甘油
|
储存缓冲液
| 20mM Tris-HCl（pH 7.4），1mM CaCl₂，50%甘油
|
保存
| 4℃，1年
|
| 注意事项 | 打开包装使用后，若在2～8℃环境下放置超过1周时间，建议过滤除菌，防止微生物污染。 |
产品用途
1. 基因诊断试剂盒、基因组DNA提取试剂盒、RNA提取试剂盒中，用于去除核酸酶和其它蛋白污染；
2. 生物制药工业中，提取核酸、多糖类药物等非蛋白类生物制品，例如DNA疫苗和肝素的制备；3. 日用工业领域，用于皮肤保健去除老化角质层细胞，以及鞣皮、制备水解蛋白调味品、化妆品及饲料加工等。
使用方法
取20-40 mg/mL的溶液，使其在消化液或裂解液中的终浓度为50-200 μg/mL；根据需求不同，作用时间可为30分钟-18小时不等，长时间反应会出现蛋白酶K自我降解现象。
酶活及单位定义
酶活：≥1800 U/mL
单位活性定义：在37℃、pH 7.5条件下，每分钟可水解酪蛋白底物生成1 μmol酪氨酸的蛋白酶K的量，定义为一个单位（U）。
最适条件与影响因素
最适pH范围
| 7.5-11.5
|
最适温度范围
| 推荐温度范围是37-70℃，其中70℃时活性最高
|
激活剂
| 1-5 mM Ca²⁺
|
抑制剂
| 二异丙基氟磷酸（DIFP）、苯甲基磺酰氟（PMSF）
|
灭活方法
| 向反应体系中加入5 mM的PMSF即可使蛋白酶K失活 |
杂质残留
本产品不含DNA和RNA，不含DNase和RNase；
以λDNA为底物于37℃消化3小时以上，未检测到脱氧核糖核酸酶活性；
以RNA为底物于37℃消化3小时以上，未检测到核糖核酸酶活性。
注意事项
1. 为保障您的安全和健康，使用时请穿戴好实验服，佩戴防护手套和护目镜后进行操作。
2. 该产品可能引发皮肤过敏反应，请谨慎操作。
3. 若蛋白酶K溶液需反复使用，需分装保存，避免污染和活性下降。
This product is derived from Pichia pastoris expressing a site-directed mutagenesis-optimized gene for Proteinase K from Engyodontium album. It offers advantages such as higher specific activity, higher yield, and a broader active pH/temperature range compared to the native enzyme. Advanced protein engineering techniques and a 28,000-liter yeast fermentation platform effectively reduce unit costs, providing a cost-effective solution for biopharmaceutical R&D and production. Yeast-based high-efficiency expression and chromatographic purification ensure product purity. This product is free from bacterial endotoxins, resulting in a broader range of applications compared to native Proteinase K.
Proteinase K is one of the most active proteases available, with an active pH range of 4-12 and a temperature range of 0-75°C. It retains activity even in the presence of SDS, Urea, or EDTA. It cleaves peptide bonds at the carboxyl side of aliphatic and aromatic amino acids and is capable of degrading all proteins.
Catalog
| P1520269
|
Product Name
| Proteinase K Solution (40 mg/mL)
|
Source/Species
| Pichia pastoris expressing a genetically engineered Engyodontium album Proteinase K gene.
|
Molecular Weight
| 29.3 kDa
|
Isoelectric Point (pI)
| 8.9
|
Dilution Buffer
| 20 mM Tris-HCl (pH 7.4), 1 mM CaCl₂; or 20 mM Tris-HCl (pH 7.4), 1 mM CaCl₂, 2% Glycerol.
|
Storage Buffer
| 20 mM Tris-HCl (pH 7.4), 1 mM CaCl₂, 50% Glycerol.
|
Storage
| Stable for 1 year at 4°C.
|
Note
| After opening, if stored at 2–8°C for more than one week, it is recommended to filter sterilize the solution to prevent microbial contamination. |
Applications
In genetic diagnostic kits, genomic DNA extraction kits, and RNA extraction kits for removing nucleases and other protein contaminants.
In the biopharmaceutical industry for the extraction of nucleic acids, polysaccharide-based drugs, and other non-protein biological products, such as in the preparation of DNA vaccines and heparin.
In the daily chemical industry for skin care (removing aged stratum corneum cells), as well as in leather tanning, the production of hydrolyzed protein flavorings, cosmetics, and feed processing.
Usage Instructions
Use the 20-40 mg/mL stock solution to achieve a final concentration of 50-200 μg/mL in the digestion or lysis buffer. Incubation times can range from 30 minutes to 18 hours depending on the application requirements. Prolonged incubation may lead to autodegradation of the enzyme.
Enzyme Activity and Unit Definition
Optimal Conditions and Influencing Factors
Optimal pH Range
| 7.5-11.5
|
Optimal Temperature Range
| Recommended range: 37-70°C (Highest activity at 70°C)
|
Activators
| 1-5 mM Ca²⁺
|
Inhibitors
| Diisopropyl fluorophosphate (DIFP), Phenylmethylsulfonyl fluoride (PMSF)
|
Inactivation Method
| Add PMSF to the reaction system to a final concentration of 5 mM. |
Residual Impurities Testing
This product is free from DNA and RNA, and contains no detectable DNase or RNase activity.
No deoxyribonuclease activity was detected after incubating with λDNA substrate for over 3 hours at 37°C.
No ribonuclease activity was detected after incubating with an RNA substrate for over 3 hours at 37°C.
Precautions
To ensure your safety and health, please wear appropriate laboratory attire, protective gloves, and goggles during operation.
This product may cause skin allergic reactions; handle with care.
If the Proteinase K solution requires repeated use, aliquot it to avoid contamination and loss of activity.
ZJ26F0838623
危险品化学品经营许可证（带存储）