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Volume 151, Issue 11

Protein -mannosyltransferase A of is involved in -mannosylation of glucoamylase I Free

Abstract

Industrially important extracellular enzymes from filamentous fungi are often -mannosylated. The structure and function of the () gene encoding the protein --mannosyltransferase of were characterized. The disruptant, designated AaPMTA, was constructed by homologous recombination. The strain AaPMTA exhibited fragile cell morphology with respect to hyphal extension, as well as swollen hyphae formation and conidia formation in potato dextrose medium. Moreover, the disruptant showed increased sensitivity to high temperature and Congo red. Thus, the AaPmtA protein is involved in the formation of the normal cell wall. The strain AaPMTA could grow well in liquid synthetic medium and secrete glucoamylase I (GAI-AaPMTA) to a similar extent to the wild-type strain (GAI-WT). Matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis of the GAIs revealed that approximately 33 mannose moieties of GAI were absent in strain AaPMTA. This result indicates that the AaPmtA protein is responsible for the transfer of mannose to GAI. Structural analysis of the -linked oligosaccharides of GAI also demonstrated that the disruption resulted in a reduction of the amounts of -linked oligosaccharides, such as -mannose and -1,2-mannotriose, in GAI-AaPMTA. However, the amount of -1,2-mannobiose was comparable between GAI-WT and GAI-AaPMTA. The result suggests the presence of a compensatory mechanism in the synthetic pathway of -mannosylation in .

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Keyword(s): CD, circular dichroism , GAI, glucoamylase I , GlcNAc, N-acetylglucosamine , MALDI-TOF, matrix-assisted laser desorption ionization time-of-flight , Pmt, protein O-d-mannosyltransferase and wt, wild-type
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2005-11-01
2024-04-19
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Protein O-mannosyltransferase A of Aspergillus awamori is involved in O-mannosylation of glucoamylase I
Microbiology 151, 3657 (2005); https://doi.org/10.1099/mic.0.28088-0
/content/journal/micro/10.1099/mic.0.28088-0
/content/journal/micro/10.1099/mic.0.28088-0
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