title: Novel ciliate lipases for enzyme replacement during exocrine pancreatic insufficiency.
abstract: OBJECTIVE
  Exocrine pancreatic insufficiency caused by inflammation or pancreatic tumors results in nutrient malfunction by a lack of digestive enzymes and neutralization compounds. Despite satisfactory clinical results with current enzyme therapies, a normalization of fat absorption in patients is rare. An individualized therapy is required that includes high dosage of enzymatic units, usage of enteric coating, and addition of gastric proton pump inhibitors. The key goal to improve this therapy is to identify digestive enzymes with high activity and stability in the gastrointestinal tract.
  
  
  METHODS
  We cloned and analyzed three novel ciliate lipases derived from Tetrahymena thermophila. Using highly precise pH-STAT-titration and colorimetric methods, we determined stability and lipolytic activity under physiological conditions in comparison with commercially available porcine and fungal digestive enzyme preparations. We measured from pH 2.0 to 9.0, with different bile salts concentrations, and substrates such as olive oil and fat derived from pig diet.
  
  
  RESULTS
  Ciliate lipases CL-120, CL-130, and CL-230 showed activities up to 220-fold higher than Creon, pancreatin standard, and rizolipase Nortase within a pH range from pH 2.0 to 9.0. They are highly active in the presence of bile salts and complex pig diet substrate, and more stable after incubation in human gastric juice compared with porcine pancreatic lipase and rizolipase.
  
  
  CONCLUSIONS
  The newly cloned and characterized lipases fulfilled all requirements for high activity under physiological conditions. These novel enzymes are therefore promising candidates for an improved enzyme replacement therapy for exocrine pancreatic insufficiency.
authors: aBiomedical Technology Center of the Medical Faculty, University of Muenster bCilian AG, Muenster cDepartment of Surgery, St Josef-Hospital Bochum, Ruhr-University Bochum, Bochum, Germany.
authors: Alexander
authors: A
authors: Brock
authors: Ingo
authors: I
authors: Aldag
authors: Stella
authors: S
authors: Edskes
authors: Marcus
authors: M
authors: Hartmann
authors: Torsten
authors: T
authors: Herzog
authors: Waldemar
authors: W
authors: Uhl
authors: Juergen
authors: J
authors: Schnekenburger
mesh_terms: D000681:Amylases / Q000737:chemistry; D000821:Animal Feed; D000818:Animals; D001647:Bile Acids and Salts; D003001:Cloning, Molecular / Q000379:methods; D003124:Colorimetry / Q000379:methods; D004338:Drug Combinations; D010450:Endopeptidases / Q000737:chemistry; D056947:Enzyme Replacement Therapy / Q000379:methods*; D010188:Exocrine Pancreatic Insufficiency / Q000188:drug therapy*; D005285:Fermentation; D005750:Gastric Juice / Q000201:enzymology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000737:chemistry* / Q000235:genetics; D008066:Lipolysis; D020799:Pancrelipase / Q000737:chemistry; D034421:Sus scrofa; D016808:Tetrahymena thermophila / Q000201:enzymology
chemical_list: D001647:Bile Acids and Salts; D004338:Drug Combinations; D020799:Pancrelipase; C059312:nortase; D008049:Lipase; D000681:Amylases; D010450:Endopeptidases