Abstract
We purified an 18.8 kD protease from caricain solution. This protease was derived from caricain. It does not have the first 41 residues of the N-terminal sequence of caricain, and its N-terminal residue is Thr. Also, one of the disulfide bonds of caricain (cys22–cys63) was opened during the formation of the protease. We named this 18.8 kD protease caricain II. Caricain II has a wide pH range, and it is more sensitive to temperature changes than caricain. The proteolytic activity of caricain II is twice as much as that of caricain using casein as a substrate. However, caricain II has a low hydrolytic activity with N-benzoyl-l-arginine ethyl ester (BAEE) that is one of the special substrates of caricain. Our results indicate that caricain II is remarkably different from caricain and it can provide an improvement over caricain on the proteolytic activity.
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Abbreviations
- A:
-
alanine
- BAEE:
-
N-benzoyl-l-arginine ethyl ester
- C or Cys:
-
cysteine
- CM Cellulose:
-
carboxymethyl cellulose
- D:
-
aspartic acid
- E or Glu:
-
glutamic acid
- EDTA:
-
ethylene diamine tetra-acetic acid
- F:
-
phenylalanine
- G or Gly:
-
glycine
- H:
-
histidine
- HCl:
-
hydrochloride
- I:
-
isoleucine
- K or Lys:
-
lysine
- L or Leu:
-
leucine
- M:
-
methionine
- N:
-
asparagine
- NaCl:
-
sodium chloride
- NaOH:
-
sodium hydroxide
- (NH4)2SO4:
-
ammonlumsulfate
- -NH2:
-
amino group
- OD value:
-
optical density value
- P:
-
proline
- PDS:
-
2,2′-dipyridyl disulfide
- PVDF:
-
polyvinylidene difluoride
- Q:
-
glutamine
- R:
-
arginine
- S:
-
serine
- SDS-PAGE:
-
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- SH:
-
sulfhydryl group
- T or Thr:
-
threonine
- Tris:
-
tris (Hydroxymethyl) aminomethane
- V or Val:
-
valine
- W:
-
tryptophan
- Y:
-
tyrosine.
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Liu, W., Ye, W., Wang, Z. et al. Preparation and Characterization of a Truncated Caricain Lacking 41 Residues from the N-terminal. Protein J 24, 243–251 (2005). https://doi.org/10.1007/s10930-005-6720-5
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DOI: https://doi.org/10.1007/s10930-005-6720-5