Regulatory mechanisms and cell membrane properties of Candida glycerinogenes differ under 2‐phenylethanol addition or fermentation conditions

Author:

Wang Yuqin1234,Liu Fang134,Lu Xinyao134,Zong Hong134,Zhuge Bin134ORCID

Affiliation:

1. The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology Jiangnan University Wuxi China

2. School of Grain Science and Technology Jiangsu University of Science and Technology Zhenjiang China

3. The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology Jiangnan University Wuxi China

4. Research Centre of Industrial Microbiology, School of Biotechnology Jiangnan University Wuxi China

Abstract

AbstractThe biosynthesis of 2‐phenylethanol (2‐PE) at high yields and titers is often limited by its toxicity. In this study, we describe the molecular mechanisms of 2‐PE tolerance in the multi‐stress tolerant industrial yeast, Candida glycerinogenes. They were different under 2‐PE addition or fermentation conditions. After extracellular addition of 2‐PE, C. glycerinogenes cells became rounder and bigger, which reduced specific surface area. However, during 2‐PE fermentation C. glycerinogenes cells were smaller, which increased specific surface area. Other differences in the tolerance mechanisms were studied by analyzing the composition and molecular parameters of the cell membrane. Extracellular 2‐PE stress resulted in down‐regulation of transcriptional expression of unsaturated fatty acid synthesis genes. This raised the proportion of saturated fatty acids in the cell membrane, which increased rigidity of the cell membrane and reduced 2‐PE entry to the cell. However, intracellular 2‐PE stress resulted in up‐regulation of transcriptional expression of unsaturated fatty acid synthesis genes, and increased the proportion of unsaturated fatty acids in the cell membrane; this in turn enhanced flexibility of the cell membrane which accelerated efflux of 2‐PE. These contrasting mechanisms are mediated by transcriptional factors Hog1 and Swi5. Under 2‐PE addition, C. glycerinogenes activated Hog1 and repressed Swi5 to upregulate erg5 and erg4 expression, which increased cell membrane rigidity and resisted 2‐PE import. During 2‐PE fermentation, C. glycerinogenes activated Hog1 and repressed Swi5 to upregulate 2‐PE transporter proteins cdr1 and Acyl‐CoA desaturase 1 ole1 to increase 2‐PE export, thus reducing 2‐PE intracellular toxicity. The results provide new insights into 2‐PE tolerance mechanisms at the cell membrane level and suggest a novel strategy to improve 2‐PE production by engineering anti‐stress genes.

Funder

National Natural Science Foundation of China

Publisher

Wiley

Subject

Molecular Medicine,Applied Microbiology and Biotechnology,General Medicine

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