蛋白酶 K 来源于林伯氏白色念球菌

3.4.21.64 ( BRENDA | IUBMB )

MFCD00132129

Purified Proteinase K DNA free SAE0151 undergoes strict quality control testing to ensure the absence of detectable levels of contaminating DNA using 35 cycles PCR amplification of 16S and 18S rDNA using universal primer sets.

Proteinase K is a stable serine protease with broad substrate specificity. It degrades many proteins in the native state even in the presence of detergents. Proteinase K was isolated from a fungus able to grow on keratin and the enzyme can digest native keratin. The enzyme belongs to the subtilisin family with an active site catalytic triad (Asp39 -His69 -Ser224).
The predominant site of cleavage is the peptide bond adjacent to the carboxyl group of aliphatic and aromatic amino acids with blocked alpha amino groups. It is commonly used for its broad specificity. Proteinase K is usually denatured by subsequent phenol extractions.

The study of microbial communities has been revolutionized in recent years by the widespread adoption of culture independent analytical techniques such as 16S rRNA gene sequencing and metagenomics. Since DNA contamination during sample preparation is a major problem of these sequence-based approaches, DNA extraction reagents free of DNA contaminates are essential.

在分离DNA和RNA的过程中，用于核酸酶的蛋白水解失活。
用于去除内毒素结合的阳离子蛋白，如溶菌酶和核糖核酸酶A。
据报道，其可用于分离肝、酵母和绿豆线粒体
用于测定酶在细胞膜上的定位
用于石蜡包埋组织切片的处理，暴露用于抗体标记的抗原结合位点。
用于可传播海绵状脑病(TSE)研究中朊病毒脑组织样本蛋白的消化。

• Proteinase K is frequently used in molecular biology applications to digest unwanted proteins, such as nucleases from DNA or RNA preparations from microorganisms, cultured cells, and plants.
• Removes endotoxins that bind to cationic proteins such as lysozyme and ribonuclease A useful for the isolation of hepatic, yeast, and mung bean mitochondria.
• Determination of enzyme localization on membranes
• Treatment of paraffin embedded tissue sections to expose antigen binding sites for antibody labeling.
• Digestion of proteins from brain tissue samples for prions in Transmissible Spongiform Encephalopathies (TSE) research.

蛋白酶K是一种稳定且具有高活性的丝氨酸蛋白酶。晶体和分子结构研究证据表明该酶属于枯草素家族，具有活性位点催化三联体(Asp³⁹-His⁶⁹-Ser²²⁴)。它在一系列广泛的环境中均稳定：pH，缓冲盐，去污剂(SDS)以及温度。 在0.1-0.5% SDS存在的情况下，蛋白酶K可保持活性，在不损害分离DNA完整性的前提下，可以消化DNA制剂中的多种蛋白质和核酸酶。

Purified Proteinase K DNA free SAE0151 undergoes strict quality control testing to ensure it will be free of DNA contaminants, suitable for Microbiome research.

One unit will hydrolyze urea denatured hemoglobin to produce color equivalent to 1.0 micromole of tyrosine per minute at pH 7.5, 37℃ (color per Folin-Ciocalteu reagent).

Proteinase K is active in 1% Triton X-100 and in 0.5% (w/v) SDS. SDS and urea will denature protein substrates resulting in increased digestion rates. Proteinase K itself is denatured much more slowly by these agents.

200

30 units/mg protein

28.93 kDa

DNA free

wet ice

−20℃

GHS08

Danger

H315 - H319 - H334

P261 - P264 - P280 - P302 + P352 - P304 + P340 + P312 - P305 + P351 + P338

Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2

11 - Combustible Solids

WGK 1