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Characterization of an extracellularly derived α-mannosidase from the liquid exudate of the sclerotia of Sclerotinia sclerotiorum (Lib.) de Bary

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Abstract

Class I α-mannosidases play an important role in co- and post-translational N-glycosylation modification of proteins, and also in glycoprotein glycan hydrolysis. Herein, we investigated a protein named Man-41, from liquid exudate droplets secreted on the surface of developing sclerotia by Sclerotinia sclerotiorum. The protein was identified by MALDI-TOF mass spectrometry to be a α-mannosidase. The full-length open reading frame of Man-41 consists of 1581 bp, encoding 526 amino acid residues and containing a putative signal peptide at amino acid residues 1–20, and a conserved sequence at residues 50–521. Man-41 was classified into glycoside hydrolase family 47 (GH47) by clustering analysis. The catalytic residues include Glu125, Arg129, Asp270, Ser271, Glu274, Arg420, Glu422, Glu425, Glu485, Thr514, and Glu515, which are conserved in all Class I α-1,2-mannosidases. Recombinant Man-41 protein had 26.67 ± 2.18 U/mg of α-mannosidase activity, about one-half of intracellular mannosidase activity of sclerotia. In conclusion, this is the first time that mannosidase has been identified in an extracellular fluid and Man-41 is also a new member of GH47 with Ca2+-dependent characteristics. This work lays the foundation for further study of the function of Man-41 in sclerotial development.

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Acknowledgments

We thank Hong Jin from Fudan University for TOF–MS analysis and Di Guan from Hunan Academy of Agricultural Sciences for ICP-MS analysis. This work was supported by the National 973 Project of China (2012CB910501) and the National Natural Science Foundation of China (31071455).

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    Authors and Affiliations

    1. Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Hunan Agricultural University, Changsha, 410128, Hunan, China

      Zhengli Liu, Ran Wei, Wen He, Ying Ruan & Chunlin Liu

    2. College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha, 410128, Hunan, China

      Ying Ruan

    3. Agricultural College, Hunan Agricultural University, Changsha, 410128, Hunan, China

      Zhengli Liu, Ran Wei, Wen He & Chunlin Liu

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    1. Zhengli Liu
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    Correspondence to Chunlin Liu.

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    Zhengli Liu and Ran Wei have contributed equally to this work.

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    Liu, Z., Wei, R., He, W. et al. Characterization of an extracellularly derived α-mannosidase from the liquid exudate of the sclerotia of Sclerotinia sclerotiorum (Lib.) de Bary. Antonie van Leeuwenhoek 108, 107–115 (2015). https://doi.org/10.1007/s10482-015-0468-5

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