Alexandre H, Blanchet S, Charpentier C
Laboratoire d'oenologie, Institut Jules Guyot, Université de Bourgogne, BP 27877, 21078, Dijon, France.
FEMS Microbiol Lett. 2000 Apr 15;185(2):147-50. doi: 10.1111/j.1574-6968.2000.tb09053.x.
Analysis of velum-forming yeast cell wall components released by beta-1,3-glucanase treatment were compared with those of a non velum-forming yeast. SDS-PAGE electrophoresis and Western blotting with ConA-peroxidase staining of mannoproteins allowed us to identify a 49-kDa mannoprotein present in the cell wall of the velum-forming yeast and hardly visible in the control. The cell wall nature of this protein was confirmed by labelling with the non-permeable sulfosuccinimydiyl-6-(biotinamido)hexanoate reagent. A partial purification of this mannoprotein by anion exchange HPLC followed by surface hydrophobicity determination revealed that the fraction containing the 49-kDa mannoprotein was the most hydrophobic. Since cell surface hydrophobicity plays an important role in aggregate formation, it is likely that this mannoprotein is involved in velum formation.
将经β-1,3-葡聚糖酶处理释放的形成菌膜酵母细胞壁成分与非形成菌膜酵母的细胞壁成分进行了比较。通过SDS-PAGE电泳和用ConA-过氧化物酶对甘露糖蛋白进行染色的Western印迹法,我们鉴定出一种49 kDa的甘露糖蛋白存在于形成菌膜酵母的细胞壁中,而在对照中几乎不可见。用非渗透性的磺基琥珀酰亚胺基-6-(生物素酰胺基)己酸试剂标记证实了该蛋白的细胞壁性质。通过阴离子交换HPLC对该甘露糖蛋白进行部分纯化,随后测定表面疏水性,结果表明含有49 kDa甘露糖蛋白的级分疏水性最强。由于细胞表面疏水性在聚集体形成中起重要作用,这种甘露糖蛋白很可能参与了菌膜的形成。