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. 2022 Nov 21;11(22):3744.
doi: 10.3390/foods11223744.

Effects on Cell Membrane Integrity of Pichia anomala by the Accumulating Excessive Reactive Oxygen Species under Ethanol Stress

Yanru Chen  1   2 , Yin Wan  1   2 , Wenqin Cai  1   2 , Na Liu  1   2 , Jiali Zeng  1   2 , Chengmei Liu  1   2 , Hong Peng  1   2 , Guiming Fu  1   2
Affiliations

Effects on Cell Membrane Integrity of Pichia anomala by the Accumulating Excessive Reactive Oxygen Species under Ethanol Stress

Yanru Chen et al. Foods. .

Abstract

Ethanol stress to yeast is well recognized and exists widely during the brewing process of alcohol products. Pichia anomala is an important ester-producing yeast in the brewing process of Chinese Baijiu and other alcohol products. Therefore, it is of great significance for the alcohol products brewing industry to explore the effects of ethanol stress on the growth metabolism of P. anomala. In this study, the effects of ethanol stress on the growth, esters production ability, cell membrane integrity and reactive oxygen species (ROS) metabolism of P. anomala NCU003 were studied. Our results showed that ethanol stress could inhibit the growth, reduce the ability of non-ethyl ester compounds production and destroy the cell morphology of P. anomala NCU003. The results also showed that 9% ethanol stress produced excessive ROS and then increased the activities of antioxidant enzymes (superoxide dismutase, catalase, aseorbateperoxidase and glutathione reductase) compared to the control group. However, these increased antioxidant enzyme activities could not prevent the damage caused by ROS to P. anomala NCU003. Of note, correlation results indicated that high content of ROS could promote the accumulation of malondialdehyde content, resulting in destruction of the integrity of the cell membrane and leading to the leakage of intracellular nutrients (soluble sugar and protein) and electrolytes. These results indicated that the growth and the non-ethyl ester compounds production ability of P. anomala could be inhibited under ethanol stress by accumulating excessive ROS and the destruction of cell membrane integrity in P. anomala.

Keywords: Pichia anomala; cell membrane integrity; ester production ability; ethanol stress; reactive oxygen species.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Effects of ethanol stress on the growth of P. anomala NCU003 in liquid medium (A) and solid medium (B).
Figure 2
Figure 2
Effects of ethanol stress on the content of ethyl acetate (A), ethyl propanoate (B), ethyl hexanoate (C), ethyl caprylate (D), ethyl caprate (E), isoamyl formate (F), phenylethyl acetate (G), isopentyl acetate (H) and isobutyl acetate (I) of P. anomala NCU003.
Figure 3
Figure 3
SEM images of P. anomala NCU003 under different ethanol stress concentrations ((A), (a): control; (B), (b): 3% ethanol; (C), (c): 6% ethanol; (D), (d): 9% ethanol; (A–D): 10×; (a–d): 20×).
Figure 4
Figure 4
The PI staining results of P. anomala NCU003 under different ethanol stress concentrations ((A), (a): control; (B), (b): 3% ethanol; (C), (c): 6% ethanol; (D), (d): 9% ethanol).
Figure 5
Figure 5
Effects of ethanol stress on ROS content of P. anomala NCU003 (different alphabet represents a significant difference (p < 0.05) at the same time).
Figure 6
Figure 6
Correlation analysis of cell membrane integrity and ROS-metabolism-related indicators of the control (A) and 9% ethanol stress (B) groups in P. anomala NCU003 (EC: electrical conductivity; SS: soluble sugar; SP: soluble protein; *: p < 0.05; **: p < 0.05; ***: p < 0.001).
Figure 7
Figure 7
Schematic diagram of the influence mechanism of ethanol stress on P. anomala.
See this image and copyright information in PMC

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