Characteristics and determination of neuraminidase from Clostridium perfringens in Worthington

AI Meijie Worthington Neutral protease （ Dispersing enzyme ） background ：

Neutral protease  (Dispase)  Is a non mammalian source of non  (AOF)  Metalloproteinase , adopt  Worthington  Developed method of purification . Its mild proteolysis makes the enzyme particularly suitable for the preparation of primary cells and secondary （ Pass it on from generation to generation ） Cell culture , Because it is mild to the cell membrane . The protease is also used as an auxiliary enzyme in cell separation and tissue dissociation applications , Usually used with collagenase .

 

 

Worthington  Bacillus polymyxa neutral protease  (Dispase)  Characteristics of ：

Molecular weight ：  36 kda.

The best  pH： In a very wide  pH  Stable in range ：4.0-9.0, The best  pH 5.9-7.0.

stability / Store ： stay  2-8°C  Lower stability  12  Months . After re dissolving with water or commonly used balanced salt solution or culture medium , Repackage and store in  -20°C.

Unit definition ： stay  37°C、pH 7.5  Under the condition of , Take casein as substrate , A unit releases  1  Micromore  Folin  Positive amino acids （ In tyrosine ）.

Specificity ： Nonspecific cleavage of peptide bonds containing leucine and phenylalanine .

Activator ： Divalent cation , Include Ca 2+、Mg 2+、Mn 2+ and Fe 2+

Inhibitors ：EDTA (1mM)、EGTA、1-10- Phenanthroline and heavy metals

application ： It is usually used to separate skin epidermis and dermis , Leave intact epithelium and stem cells 、 Isolation and application of hepatocytes and other cells . However , Due to the diversity of variables involved , It should be based on each cell / The experience of organization application determines the exact separation conditions .

Worthington Neutral protease （ Dispersing enzyme ） Related research ：

Collagenase  (CLS1-4/CLSS1-4/CLSPA)

Deoxyribonuclease  I (DP/D/DCLS/D2/DPFF/DPRF)

Elastase  (ES/ESL/ESFF)

Hyaluronidase  (HSE/HSEP)

Papain  (PAP/PAPL/PAP2)

protease K (PROK)

Trypsin  (TL/TRL/TRL3/TRLS/TRTPCK)

Trypsin inhibitor  (LBI/OI/SI/SIC)

Cell separation optimization system  (CIT)

Hepatocyte isolation system  (HIS)

Neonatal cardiomyocyte isolation system  (NCIS)

Papain dissociation system  (PDS/PDS2)

Related literature ：

1. Keratinocyte culture , Basic cell culture program ,（Pollard, J.  and  Walker, J., Eds.）,Humana Press,117,1997

2.Akimoto, Y.、Obinata, A.、Endo, H.  and  Hirano, H.： By using the epidermis of cultured chicken embryo skin - Immunohistochemical study of basement membrane reconstruction by dermal recombination experiment ： Induction of tenascin , J Histochem Cytochem 40 , 1129 , 1992

3.Alvarez, V.、von der Weid, I.、Seldin, L.  and  Santos, A.： Growth condition pair Paenibacillus peoriae NRRL BD-62  and Paenibac illus polymyxa SCE2  Effects of extracellular proteolytic enzymes . , Lett Appl  microorganism  43 , 625-30, 2006