Scorpion digestive lipase: a member of a new invertebrate's lipase group presenting novel characteristics.
Unlike classical digestive lipases, the scorpion digestive lipase (SDL) has a strong basic character. The SDL activity's optimal pH, when using tributyrin or olive oil as substrate, was 9.0. Added to that, the estimated isoelectric point of the native SDL using the electrofocusing technique, was found to be higher than 9.6. To our knowledge, this is the first report of an animal digestive lipase having such a basic character. When olive oil was used as substrate, SDL was shown to be insensitive to the presence of amphiphilic proteins such as bovine serum albumin (BSA). Furthermore, the hydrolysis was found to be specifically dependent on the presence of Ca(2+) ions, since no significant SDL activity was detected in the presence of ions chelator such as EDTA. Nevertheless, the SDL does not require Ca(2+) to trigger the hydrolysis of tributyrin emulsion. Interestingly Zn(2+) and Cu(2+) ions act as strong inhibitors of SDL activity when using tributyrin as substrate. An internal chymotryptic cleavage of SDL generated two fragments of 28 and 25 kDa having the same N-terminal sequence. This sequence of 19 residues does not share any homology with known animal and microbial lipases. Polyclonal antibodies directed against SDL (pAbs anti-SDL) failed to recognise ostrich pancreatic and dog gastric lipases (OPL and rDGL). Moreover, both pAbs anti-OPL and anti-rDGL failed to immunoreact with SDL. These immunological as well as distinct biochemical properties strengthen the idea that SDL appears to belong to a new invertebrate's lipase group.
Zouari
Nacim
N
Laboratoire de Biochimie et de Génie Enzymatique des Lipases, ENIS, BPW, 3038 Sfax, Tunisia.
Miled
Nabil
N
Rouis
Souad
S
Gargouri
Youssef
Y
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002417:Cattle; D003300:Copper / Q000494:pharmacology; D004285:Dogs; D004789:Enzyme Activation / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D043143:Hepatopancreas / Q000201:enzymology; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis / Q000187:drug effects; D007150:Immunohistochemistry; D007526:Isoelectric Point; D008049:Lipase / Q000737:chemistry / Q000378:metabolism*; D017830:Octoxynol / Q000494:pharmacology; D000069463:Olive Oil; D010938:Plant Oils / Q000378:metabolism; D012605:Scorpions / Q000201:enzymology*; D012621:Seasons; D012710:Serum Albumin, Bovine / Q000378:metabolism; D020310:Struthioniformes; D013379:Substrate Specificity; D014280:Triglycerides / Q000378:metabolism; D015032:Zinc / Q000494:pharmacology
D000069463:Olive Oil; D010938:Plant Oils; D014280:Triglycerides; D012710:Serum Albumin, Bovine; D003300:Copper; D017830:Octoxynol; D008049:Lipase; D015032:Zinc; C005830:tributyrin; D002118:Calcium