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对具有降低的巴斯德效应的酿酒酵母进行基于¹³C的代谢通量分析。

¹³C-based metabolic flux analysis of Saccharomyces cerevisiae with a reduced Crabtree effect.

作者信息

Kajihata Shuichi, Matsuda Fumio, Yoshimi Mika, Hayakawa Kenshi, Furusawa Chikara, Kanda Akihisa, Shimizu Hiroshi

机构信息

Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka 565-0871, Japan.

KANEKA Fundamental Technology Research Alliance Laboratories, Graduate School of Engineering, Osaka University, 2-8 Yamadaoka, Suita, Osaka 565-0871, Japan.

出版信息

J Biosci Bioeng. 2015 Aug;120(2):140-4. doi: 10.1016/j.jbiosc.2014.12.014. Epub 2015 Jan 26.

Abstract

Saccharomyces cerevisiae shows a Crabtree effect that produces ethanol in a high glucose concentration even under fully aerobic condition. For efficient production of cake yeast or compressed yeast for baking, ethanol by-production is not desired since glucose limited chemostat or fed-batch cultivations are performed to suppress the Crabtree effect. In this study, the (13)C-based metabolic flux analysis ((13)C-MFA) was performed for the S288C derived S. cerevisiae strain to characterize a metabolic state under the reduced Crabtree effect. S. cerevisiae cells were cultured at a low dilution rate (0.1 h(-1)) under the glucose-limited chemostat condition. The estimated metabolic flux distribution showed that the acetyl-CoA in mitochondria was mainly produced from pyruvate by pyruvate dehydrogenase (PDH) reaction and that the level of the metabolic flux through the pentose phosphate pathway was much higher than that of the Embden-Meyerhof-Parnas pathway, which contributes to high biomass yield at low dilution rate by supplying NADPH required for cell growth.

摘要

酿酒酵母表现出一种克勒勃屈利效应,即在高葡萄糖浓度下,即使在完全有氧条件下也会产生乙醇。为了高效生产用于烘焙的压榨酵母或活性干酵母,由于进行葡萄糖受限的恒化器培养或补料分批培养以抑制克勒勃屈利效应,所以不希望有乙醇副产物。在本研究中,对源自S288C的酿酒酵母菌株进行了基于¹³C的代谢通量分析(¹³C-MFA),以表征在克勒勃屈利效应降低情况下的代谢状态。酿酒酵母细胞在葡萄糖受限的恒化器条件下以低稀释率(0.1 h⁻¹)进行培养。估计的代谢通量分布表明,线粒体中的乙酰辅酶A主要由丙酮酸通过丙酮酸脱氢酶(PDH)反应产生,并且通过磷酸戊糖途径的代谢通量水平远高于糖酵解途径,这通过提供细胞生长所需的NADPH有助于在低稀释率下获得高生物量产量。

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