Effects of Supercritical CO2 Combined with Ginger Essential Oil on the Sterilization of Vibrio parahaemolyticus and Its Mechanism

doi:10.13560/j.cnki.biotech.bull.1985.2022-1207

Abstract

Abstract:

The objective is to optimize the low pressure and low temperature sterilization technology of supercritical CO₂ and to reduce the influence of bacterial quorum sensing on the sterilization effect. The LC-MS and phenol sulfuric acid were used to determine the contents of quorum sensing signal molecules and exopolysaccharides of Vibrio parahaemolyticus, respectively. The micro diffusion method was applied to determine the quorum sensing inhibitory activity of selected quorum sensing inhibitor ginger essential oil, and the combined supercritical CO₂ sterilization process was optimized. The quorum sensing signal molecules and exopolysaccharides accumulated in the bacterial suspension with the increase of the total number of colonies, activating the expression of related genes, thereby enhancing cell sensing, leading to the decline of the bactericidal effect of supercritical CO₂. Ginger essential oil can be used as quorum sensing inhibitor, and the minimum inhibitory concentration was 200 μg/mL, and it significantly inhibited the mobility of Vibrio parahaemolyticus, biofilm formation（64.20% of inhibition rate）and the production of exopolysaccharides in bacterial suspension（45.1% of inhibition rate）at the sub inhibitory concentration. The optimal process of quorum sensing inhibitor combined with supercritical CO₂ sterilization is quorum sensing inhibitor concentration 75 μg/mL, and the sterilization conditions are 15 MPa, 40℃, and 20 min. In conclusion, ginger essential oil can be used as a quorum sensing inhibitor to enhance the effect of supercritical CO₂ sterilization, providing a theoretical basis for the application of quorum sensing inhibitors.

Key words: Vibrio parahaemolyticus, quorum sensing, supercritical CO₂ sterilization, ginger essential oil

HUANG Ya-ning, ZHANG Hai-jiao, HAN Yu-qian, LIU Zun-ying. Effects of Supercritical CO₂ Combined with Ginger Essential Oil on the Sterilization of Vibrio parahaemolyticus and Its Mechanism[J]. Biotechnology Bulletin, 2023, 39(5): 297-305.

Figures/Tables 10

Table 1 Four-factor three-level orthogonal optimization table

水平 Level	因素Factor
水平 Level	压力 Pressure/MPa	温度 Temperature/℃	时间 Time/min	QSI浓度 QSI concentration/ （μg·mL^-1）
1	10	35	10	25
2	15	40	20	50
3	20	45	30	75

Fig. 1 Growth curve of bacterial solution and content variation of signal molecules The same letter indicates no statistically significant difference between the treatments（P> 0.05）, or otherwise, there is a significant difference（P <0.05）. The same below

Fig. 2 Relationship between signal molecules, extracellular polysaccharide content and initial colony concentra-tion

Fig. 3 Relationship between sterilization effect of superc-ritical CO2 and initial colony density

Fig. 4 Quorum sensing inhibitory activity of ginger esse-ntial oil against C. violaceum Control: No ginger essential oil. A: 25 μg/mL; B: 50 μg/mL; C: 75 μg/mL; D: 100 μg/mL

Fig. 5 Effect of ginger essential oil on the mobility of V. parahaemolyticus A: Control; B: 50 μg/mL; C: 100 μg/mL

Fig. 6 Effect of ginger essential oil on biofilm formation of V. parahaemolyticus

Fig. 7 Effect of ginger essential oil on extracellular polysa-ccharide content of V. parahaemolyticus suspension

Table 2 Four-factor three-level orthogonal optimization design and results

实验序号 Experiment No.	因素Factor				杀菌对数值 Sterilization logarithmic value	FDA规定的最低程度 Minimum required by FDA
实验序号 Experiment No.	压力Pressure/ MPa	温度Tempe-rature/℃	时间 Time/min	QSI浓度 QSI concentration/（μg·mL^-1）	杀菌对数值 Sterilization logarithmic value	FDA规定的最低程度 Minimum required by FDA
1	1	1	1	1	4.58	-1.42
2	1	2	2	2	5.77	-0.23
3	1	3	3	3	7.40	+1.40
4	2	1	2	3	7.43	+1.43
5	2	2	3	1	5.93	-0.07
6	2	3	1	2	5.70	-0.30
7	3	1	3	2	6.63	+0.63
8	3	2	1	3	7.46	+1.46
9	3	3	2	1	5.95	-0.05
K1	5.917	5.880	5.913	5.487
K2	6.353	6.387	6.383	5.700
K3	6.347	6.350	6.320	7.430
R	0.436	0.507	0.470	1.943
主次顺序Primary and secondary order 最优组合Optimal combination			D>A≈B≈C A₂B₂C₂D₃

Table 3 Analysis of variance of sterilization logarithm

方差来源 Source of variance	偏差平方和 Sum of squares of deviations	自由度 Freedom	F值 F value	显著性 Significance
压力Pressure	0.373	2	0.834
温度Temperature	0.479 0.390	2 2	1.061 0.865
时间Time	6.815	2	15.084	*
QSI浓度QSI concentration	8.06	8
误差Error

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Effects of Supercritical CO₂ Combined with Ginger Essential Oil on the Sterilization of Vibrio parahaemolyticus and Its Mechanism

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