PMID:15834794
| Citation |
Lee, S, Kim, T, Stahl, CH and Lei, XG (2005) Expression of Escherichia coli AppA2 phytase in four yeast systems. Biotechnol. Lett. 27:327-34 |
|---|---|
| Abstract |
To develop an effective fermentation system for producing Escherichia coliphytase AppA2, we expressed the enzyme in three inducible yeast systems: Saccharomyces cerevisiae (pYES2), Schizosaccharomyces pombe (pDS472a), and Pichia pastoris (pPICZ alphaA), and one constitutive system: P. pastoris (pGAPZalphaA). All four systems produced an extracellular functional AppA2 phytase with apparent molecular masses ranging from 51.5 to 56 kDa. During 8-day batch fermentation in shaking flasks, the inducible Pichia system produced the highest activity (272 units ml(-1) medium), whereas the Schizo. pombe system produced the lowest activity (2.8 units ml(-1)). The AppA2 phytase expressed in Schizo. pombe had 60-75% lower K(m)for sodium phytate and 28% higher heat-stability at 65 degrees C than that expressed in other three systems. However, all four recombinant AppA2 phytases had pH optimum at 3.5 and temperature optimum at 55 degrees C and similar efficacy in hydrolyzing phytate-phosphate from soybean meal. |
| Links |
PubMed Online version:10.1007/s10529-005-0704-6 |
| Keywords |
6-Phytase/biosynthesis; 6-Phytase/chemistry; Acid Phosphatase/biosynthesis; Acid Phosphatase/chemistry; Biotechnology/methods; Blotting, Western; Cloning, Molecular; Escherichia coli/enzymology; Escherichia coli Proteins/biosynthesis; Escherichia coli Proteins/chemistry; Fermentation; Hydrogen-Ion Concentration; Hydrolysis; Kinetics; Molecular Weight; Multienzyme Complexes/biosynthesis; Multienzyme Complexes/chemistry; Phosphates/chemistry; Phytic Acid/chemistry; Pichia/metabolism; Plasmids/metabolism; Recombinant Proteins/chemistry; Saccharomyces cerevisiae/metabolism; Schizosaccharomyces/metabolism; Soybeans/metabolism; Species Specificity; Temperature; Time Factors |
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