Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.4.21.1 (chymotrypsin)
 10,938 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A crude preparation, inhibiting trypsin (T), chymotrypsin (C) and papain (P), was isolated from the culture filtrate of Actinomyces sp. 9 by butanol extraction. Ion-exchange high-performance liquid chromatography (HPLC) of this preparation on a Mono S column resulted in the separation of two inhibitory fractions: one active against T (TI-9) and the other active against C and P (CPI-9). These two fractions, having the same inhibitory specificity, were also obtained from the crude extract by size-exclusion HPLC on a Superose 12 column. This method proved to be better in terms of the purity of the fractionated inhibitors than ion-exchange chromatography. Further purification of TI-9 and CPI-9 was achieved by using reversed-phase HPLC on a PEP RPC column. In this case, TI-9 was obtained as an apparently homogeneous peak. Studies on the physicochemical properties of the purified inhibitors showed them to be small molecules, similar in hydrophobicity, pH and thermal stability, but differing in solubility. Amino acid and spectral analyses provided the peptidic structure of TI-9 and gave a molecular weight of 1643, while parallel analyses of CPI-9 showed that it lacks the common amino acids.
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PMID:Chromatographic separation and partial characterization of microbial low-molecular-weight proteinase inhibitors. 320 56

Celiac Sprue is a multi-factorial disease characterized by an inflammatory response to ingested wheat gluten and similar proteins in rye and barley. Proline-rich gluten peptides from wheat, rye, and barley are relatively resistant to gastrointestinal digestion, and therefore persist in the intestinal lumen to elicit immunopathology in genetically susceptible individuals. In this study, we characterize the in vitro gluten detoxifying properties of a therapeutically promising prolyl endopeptidase from Myxococcus xanthus (MX PEP), and describe the development of a prototypical enteric-coated capsule containing a pharmacologically useful dose of this enzyme. A high-cell density fed-batch fermentation process was developed for overproduction of recombinant MX PEP in E. coli, yielding 0.25-0.4 g/L purified protein. A simple, scalable purification and lyophilization procedure was established that yields >95% pure, highly active and stable enzyme as a dry powder. The dry powder was blended with excipients and encapsulated in a hard gelatin capsule. The resulting capsule was enteric coated using Eudragit L30-D55 polymer coat, which provided sufficient resistance to gastric conditions (> 1 h in 0.01 M HCl, pH 2 with pepsin) and rapid release under duodenal conditions (15-30 min release in pH 6.0 in the presence of trypsin and chymotrypsin). In conjunction with pancreatic enzymes, MX PEP breaks down whole gluten into a product mixture that is virtually indistinguishable from that generated by the Flavobacterium meningosepticum (FM) PEP as judged by chromatographic assays. Competitive studies involving selected immunogenic peptides mixed with whole gluten reveal that both PEPs have a wide range of substrate specificity. Our results support further in vitro and in vivo evaluation of the MX PEP capsule as an oral therapeutic agent for Celiac Sprue patients.
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PMID:Fermentation, purification, formulation, and pharmacological evaluation of a prolyl endopeptidase from Myxococcus xanthus: implications for Celiac Sprue therapy. 1613 93