Mizutani Osamu, Shiina Matsuko, Yoshimi Akira, Sano Motoaki, Watanabe Takeshi, Yamagata Youhei, Nakajima Tasuku, Gomi Katsuya, Abe Keietsu
a Department of Enzymology, Graduate School of Agricultural Science , Tohoku University , Sendai , Japan.
b Department of Application Research , National Research Institute of Brewing , Higashi-Hiroshima , Japan.
Biosci Biotechnol Biochem. 2016 Sep;80(9):1781-91. doi: 10.1080/09168451.2016.1158632. Epub 2016 Mar 15.
Disruption of the kexB encoding a subtilisin-like processing protease in Aspergillus oryzae (ΔkexB) leads to substantial morphological defects when the cells are grown on Czapek-Dox agar plates. We previously found that the disruption of kexB causes a constitutive activation of the cell wall integrity pathway. To understand how the disruption of the kexB affects cell wall organization and components, we analyzed the cell wall of ΔkexB grown on the plates. The results revealed that both total N-acetylglucosamine content, which constitutes chitin, and chitin synthase activities were increased. Whereas total glucose content, which constitutes β-1,3-glucan and α-1,3-glucan, was decreased; this decrease was attributed to a remarkable decrease in α-1,3-glucan. Additionally, the β-1,3-glucan in the alkali-insoluble fraction of the ΔkexB showed a high degree of polymerization. These results suggested that the loss of α-1,3-glucan in the ΔkexB was compensated by increases in the chitin content and the average degree of β-1,3-glucan polymerization.
破坏米曲霉中编码枯草杆菌蛋白酶样加工蛋白酶的kexB(ΔkexB),会导致细胞在查氏-多克斯琼脂平板上生长时出现严重的形态缺陷。我们之前发现,kexB的破坏会导致细胞壁完整性途径的组成型激活。为了解kexB的破坏如何影响细胞壁的组织和成分,我们分析了平板上生长的ΔkexB的细胞壁。结果显示,构成几丁质的总N-乙酰葡糖胺含量和几丁质合酶活性均增加。而构成β-1,3-葡聚糖和α-1,3-葡聚糖的总葡萄糖含量降低;这种降低归因于α-1,3-葡聚糖显著减少。此外,ΔkexB的碱不溶性部分中的β-1,3-葡聚糖显示出高度聚合。这些结果表明,ΔkexB中α-1,3-葡聚糖的损失通过几丁质含量增加和β-1,3-葡聚糖聚合平均度增加得到了补偿。