PMID: 9532788Apr 9, 1998Paper

Overproduction of 1,2-alpha-mannosidase, a glycochain processing enzyme, by Aspergillus oryzae

Bioscience, Biotechnology, and Biochemistry
T YoshidaE Ichishima

Abstract

A recombinant strain of Aspergillus oryzae has been constructed in which 1,2-alpha-mannosidase, an intracellular glycochain processing enzyme with specificity toward 1,2-alpha-mannosidic linkages, has been overexpressed. For the construction, the N-terminal signal-encoding sequence of the 1,2-alpha-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-alpha-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-alpha-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. Man7GlcNAc2 accumulated temporarily during the degradation of Man9GlcNAc2 to Man5GlcNAc2 by fungal 1,2-alpha-mannosidase.

References

May 15, 1988·Analytical Biochemistry·N TomiyaN Takahashi
Dec 1, 1983·Gene·J TilburnR W Davies
Nov 1, 1993·Current Microbiology·Y ChibaE Ichishima
Apr 1, 1994·Glycobiology·K W MoremenA Herscovics
Oct 15, 1993·European Journal of Biochemistry·E BauseB Schmidt
Dec 1, 1995·Applied Microbiology and Biotechnology·B R LeeC Kumagai

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Citations

Mar 8, 2008·Acta Crystallographica. Section D, Biological Crystallography·Yuri D LobsanovP Lynne Howell
Apr 15, 2000·Glycobiology·Z KawarD L Jarvis
Oct 26, 2001·Bioscience, Biotechnology, and Biochemistry·Y KimuraM Kimura
Oct 31, 2000·Bioscience, Biotechnology, and Biochemistry·Y Kimura, E Kitahara
Sep 21, 2000·The Journal of Biological Chemistry·F ValleeP L Howell

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