Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Navigation/Suche
- Forscher*innen
- Institutionen
- Projekte
- Publikationen
- Partner
- Geldgeber
- Ausstattung
- Knowhow
- Forschungsfelder
"Köpfe" der aktuellen Meldungen:
Gewählte Publikation:
SHR Neuro Krebs Kardio Lipid Stoffw Microb
Hermann, M; Kietzmann, MU; Ivancic, M; Zenzmaier, C; Luiten, RGM; Skranc, W; Wubbolts, M; Winkler, M; Birner-Gruenberger, R; Pichler, H; Schwab, H.
Alternative pig liver esterase (APLE) - cloning, identification and functional expression in Pichia pastoris of a versatile new biocatalyst.
J BIOTECHNOL. 2008; 133(3): 301-310.
Doi: 10.1016/j.jbiotec.2007.10.010
Web of Science
PubMed
FullText
FullText_MUG
- Co-Autor*innen der Med Uni Graz
- Birner-Grünberger Ruth
- Altmetrics:
- Dimensions Citations:
- Plum Analytics:
- Scite (citation analytics):
- Abstract:
- Isolated from pig liver as a crude, inhomogeneous enzyme fraction, pig liver esterase (PLE) was found to metabolize a wide range of substrates; often in a highly stereoselective manner. This crude esterase preparation, however, contains several iso-enzymes at proportions varying from batch to batch. Racemic methyl-(4E)-5-chloro-2-isopropyl-4-pentenoate is cleaved enantioselectively by crude PLE, but not by recombinantly expressed gamma-isoform of PLE. Concluding that another PLE iso-enzyme must carry the relevant activity, we cloned and sequenced cDNAs of several PLE isoforms and functionally expressed them in Pichia pastoris. One novel isoform termed alternative pig liver esterase (APLE) was found to hydrolyze methyl-(2R,4E)-5-chloro-2-isopropyl-4-pentenoate in a highly stereoselective manner (E>200). When heterologously expressed and directed for secretion in P. pastoris, APLE was found to be localized in the periplasm. The presence or absence of a putative C-terminal ER retention signal did neither influence functional expression nor cellular localization. The recombinant enzyme, purified by ion exchange chromatography, had a specific activity of 36U (mg protein)(-1) towards racemic methyl-(4E)-5-chloro-2-isopropyl-4-pentenoate.
- Find related publications in this database (using NLM MeSH Indexing)
- Amino Acid Sequence -
- Animals -
- Catalysis -
- Cloning, Molecular -
- Electrophoresis, Polyacrylamide Gel -
- Esterases - chemistry
- Fatty Acids, Monounsaturated - metabolism
- Hydrolysis -
- Liver - enzymology
- Liver Extracts -
- Mass Spectrometry -
- Molecular Sequence Data -
- Peptide Fragments - chemistry
- Periplasm - metabolism
- Pichia - metabolism
- Protein Transport -
- Recombinant Proteins - chemistry
- Sequence Alignment -
- Substrate Specificity -
- Sus scrofa - metabolism
- Find related publications in this database (Keywords)
- biocatalysis
- pig liver esterase
- kinetic resolution
- heterologous expression
- Pichia pastoris
- periplasm