Graduate School of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe, Hyogo, 657-8501, Japan.
Biomass Engineering Program, RIKEN, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045, Japan.
Microb Cell Fact. 2018 Sep 25;17(1):153. doi: 10.1186/s12934-018-0998-4.
Following cellulose, chitin is the most abundant renewable resource and is composed of the monomeric amino sugar N-acetyl-D-glucosamine (GlcNAc). Although many yeasts, including Saccharomyces cerevisiae, have lost their ability to utilize GlcNAc, some yeasts are able to use GlcNAc as a carbon source. However, our understanding of the effects of GlcNAc on the intracellular metabolism of nitrogen-containing compounds in these yeast species is limited.
In the present study, we quantitatively investigated the metabolic responses to GlcNAc in the GlcNAc-assimilating yeast Scheffersomyces stipitis (formerly known as Pichia stipitis) using capillary electrophoresis time-of-flight mass spectrometry (CE-TOFMS). The comprehensive analysis of the metabolites extracted from S. stipitis cells grown in glucose, xylose, or GlcNAc revealed increased intracellular accumulation of a wide range of nitrogen-containing compounds during GlcNAc assimilation in this yeast. The levels of aromatic, branched-chain, and sulfur-containing amino acids and adenine, guanine, and cytosine nucleotides were the highest in GlcNAc-grown cells.
The CE-TOFMS analysis revealed a positive effect for GlcNAc on the intracellular concentration of a wide range of nitrogen-containing compounds. The metabolomic data gathered in this study will be useful for designing effective genetic engineering strategies to develop novel S. stipitis strains for the production of valuable nitrogen-containing compounds from GlcNAc.
继纤维素之后,几丁质是最丰富的可再生资源,由单体氨基糖 N-乙酰-D-葡萄糖胺(GlcNAc)组成。尽管许多酵母,包括酿酒酵母,已经失去了利用 GlcNAc 的能力,但有些酵母能够将 GlcNAc 用作碳源。然而,我们对 GlcNAc 对这些酵母物种含氮化合物的细胞内代谢的影响的理解是有限的。
在本研究中,我们使用毛细管电泳飞行时间质谱(CE-TOFMS)定量研究了 GlcNAc 同化酵母酿酒酵母(以前称为毕赤酵母)中 GlcNAc 的代谢反应。从在葡萄糖、木糖或 GlcNAc 中生长的 S. stipitis 细胞中提取的代谢物的综合分析表明,在这种酵母中 GlcNAc 同化过程中,广泛的含氮化合物在细胞内积累增加。芳香族、支链和含硫氨基酸以及腺嘌呤、鸟嘌呤和胞嘧啶核苷酸的水平在 GlcNAc 生长的细胞中最高。
CE-TOFMS 分析显示 GlcNAc 对广泛的含氮化合物的细胞内浓度有积极影响。本研究中收集的代谢组学数据将有助于设计有效的遗传工程策略,以开发新型 S. stipitis 菌株,从 GlcNAc 生产有价值的含氮化合物。