Overproduction of 1, 2-α-Mannosidase, a Glycochain Processing Enzyme, by Aspergillus oryzae

概要

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A recombinant strain of Aspergillus oryzae has been constructed in which 1, 2-α-mannosigase, an intracellular glycochain processing enzyme with specificity toward 1, 2-α-mannosidic linkages, has been overexoressed. For the construction, the N-terminal signal-encoding sequence of the 1, 2-α-mannosidase gene (msdC) from Penicillium citrinum was replaced with that of the aspergillopepsin I signal, and the fused gene was inserted between amyB promoter-terminator elements in the expression plasmid pTAPM1. A transformant of A. oryzae (the strain PM-1) secreted a great deal of heterogeneous 1, 2-α-mannosidase into the culture media, which was purified by CM ion-exchange chromatography. Approximately 21 mg of the purified enzyme was obtained per liter of culture. N-terminal amino acid analysis indicated that the signal peptide was removed from the secreted enzyme. The Penicillium 1, 2-α-mannosidase expressed in A. oryzae did not show any notable difference from the enzyme from P. citrinum in such properties as M_r, specific activity, CD spectra, or kinetic parameters. Man_7GlcNAc_2 accumulated temporarily during the degradation of Man_9GlcNAc_2 to Man5GlcNAc_2 by fungal 1, 2-α-mannosidase.
社団法人日本農芸化学会の論文
1998-02-23