Veenhuis M, van Dijken J P, Harder W
Arch Microbiol. 1976 Dec 1;111(1-2):123-35. doi: 10.1007/BF00446559.
The localization of methanol oxidase activity in cells of methanol-limited chemostat cultures of the yeast Hansenula polymorpha has been studied with different cytochemical staining techniques. The methods were based on enzymatic or chemical trapping of the hydrogen peroxide produced by the enzyme during aerobic incubations of whole cells in methanol-containing media. The results showed that methanol-dependent hydrogen peroxide production in either fixed or unfixed cells exclusively occurred in peroxisomes, which characteristically develop during growth of this yeast on methanol. Apart from methanol oxidase and catalase, the typical peroxisomal enzymes D-aminoacid oxidase and L-alpha-hydroxyacid oxidase were also found to be located in the peroxisomes. Urate oxidase was not detected in these organelles. Phase-contrast microscopy of living cells revealed the occurrence of peroxisomes which were cubic of form. This unusual shape was also observed in thin sections examined by electron microscopy. The contents of the peroxisomes showed, after various fixation procedures, a completely crystalline or striated substructure. It is suggested that this substructure might represent the in vivo organization structure of the peroxisomal enzymes.
利用不同的细胞化学染色技术，对多形汉逊酵母在甲醇限制恒化器培养条件下细胞中甲醇氧化酶活性的定位进行了研究。这些方法基于在含甲醇培养基中对全细胞进行需氧培养时，酶产生的过氧化氢的酶促或化学捕获。结果表明，无论是固定细胞还是未固定细胞，依赖甲醇产生过氧化氢的过程仅发生在过氧化物酶体中，过氧化物酶体是该酵母在甲醇上生长时特有的细胞器。除甲醇氧化酶和过氧化氢酶外，典型的过氧化物酶体酶D-氨基酸氧化酶和L-α-羟基酸氧化酶也位于过氧化物酶体中。在这些细胞器中未检测到尿酸氧化酶。活细胞的相差显微镜观察显示存在呈立方体形状的过氧化物酶体。在电子显微镜检查的薄片中也观察到这种不寻常的形状。经过各种固定程序后，过氧化物酶体的内容物显示出完全结晶或条纹状的亚结构。有人认为，这种亚结构可能代表过氧化物酶体酶的体内组织结构。
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