Purification and characterisation of two isozymes of pyruvate decarboxylase from Rhizopus oryzae

Date

2007-3

Author

Açar, Şeyda
Yücel, Meral
Hamamcı, Haluk

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Pyruvate decarboxylases were purified and partially characterised for the first time from aerobically grown Rhizopus oryzae mycelium. Two very similar pyruvate decarboxylase isoenzymes were partially separated from each other by a purification protocol including ammonium sulphate precipitation, gel filtration chromatography and anion exchange chromatography. The subunit molecular weights of the enzymes were estimated to be 61 kDa for both. The pI values determined by two-dimensional electrophoretic analysis were 5.94 and 5.82 for the isoenzymes. The isoenzymes were similar in many kinetic aspects and the K,,, values were estimated as 3.9 and 4.5 mM for each of the enzymes. Both of the isozymes showed sigmoidal kinetics with a Hill coefficient of 1.8. Substrate inhibition was observed for both of the enzymes at pyruvate concentrations above 20 mM for one of the isoenzymes and 40 mM for the second isoenzyme. The enzymes were found in two different native forms; dimer and tetramer with estimated molecular weights of 120-128 and 272 kDa with the dimer being the predominant form. The optimum pH was between 6.3 and 6.5 for both.

Subject Keywords

Rhizopus oryzae, Pyruvate decarboxylase, Characterisation

URI

https://hdl.handle.net/11511/28198

Journal

Enzyme and Microbial Technology

DOI

https://doi.org/10.1016/j.enzmictec.2006.05.028

Collections

Department of Food Engineering, Article

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Citation Formats

Ş. Açar, M. Yücel, and H. Hamamcı, “Purification and characterisation of two isozymes of pyruvate decarboxylase from Rhizopus oryzae,” Enzyme and Microbial Technology, pp. 675–682, 2007, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/28198.