Synergistic Effect and Mechanism of Phloretin and Lauraldehyde on the Killing of Alicyclobacillus acidoterrestris Spores

doi:10.12141/j.issn.1000-565X.240526

Abstract

Abstract:

To address the challenge of Alicyclobacillus acidoterrestris spore contamination of fruit juice, this study aimed to screen and evaluate the bactericidal effect of a synergistic combination of plant-derived actives and their mechanism of action. The study determined the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of plant-derived active compounds. By calculating the fractional inhibitory concentration index (FICI), a synergistic combination against Alicyclobacillus acidoterrestris spores was identified: phloretin (Ph) and lauric aldehyde (La). The results show that the MIC of phloretin is 0.1 mg/mL, and MBC is more than 6.4 mg/mL; the MIC of lauraldehyde is 0.4 mg/mL, and MBC is more than 6.4 mg/mL; the FICI of the combined effect of the two substances is 0.5. The 8PL (2 MIC_Ph + 2 MICLa) synergistic combination shows the best spore-killing effect in the synergistic combinations, and in the culture medium, the number of spores decreases by 10^6.1 CFU/mL after 2 days of treatment in the 8PL synergistic combination; inoculating 10⁴ CFU/mL of Alicyclobacillus acidoterrestris spores in apple juice, the 8PL synergistic combination treatment can reduce the spores to 0 after 1 day and maintain the effect for at least 14 days. For apple juice samples with an inoculation amount of 10⁶ CFU/mL, the 8PL synergistic combination treatment requires more than 3 days to reduce spores to 0 CFU/mL, but after 14 days, 10^2.2 CFU/mL spores are detected again. As the concentration of synergistic combination increases, the spore absorbance significantly decreases and the fluorescence spectrum increases. By scanning electron microscopy, irregular wrinkles on the surface of the spores disappear, and the surface gradually becomes smooth with an increase in the degree of depression and fragmentation. Transmission electron microscopy (TEM) observations revealed spore membrane disruption, accompanied by progressively increasing leakage of nucleic acids and proteins. The above results indicate that the synergistic combination will cause certain damage to structures such as spore membranes. In addition, the release of pyridine dicarboxylic acid (DPA) also significantly increases, greatly reducing the stress resistance of spores. In summary, the synergistic combination of phloretin and lauraldehyde has a good killing effect on Alicyclobacillus acidoterrestris spores, and it is an effective strategy for fruit juice antibacterial and preservative properties.

Key words: Alicyclobacillus acidoterrestris, spores, juice, phloretin, lauraldehyde, food control technology

CLC Number:

TS255.44

XIAO Xinglong, ZHANG Ziqiang, CHEN Juntai, XIE Jialin, CAO Yifang. Synergistic Effect and Mechanism of Phloretin and Lauraldehyde on the Killing of Alicyclobacillus acidoterrestris Spores[J]. Journal of South China University of Technology(Natural Science Edition), 2025, 53(10): 145-154.

Figures/Tables 8

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References 37

[1]	耿敬章．欧姆加热对苹果汁中嗜酸耐热菌的杀灭作用研究［D］．西安：陕西师范大学，2006.
[2]	马丽达．苹果汁中嗜酸耐热菌菌学特性及控制方法研究［D］．泰安：山东农业大学，2014.
[3]	YOKOTA A， FUJII T， GOTO K ． Alicyclobacillus：thermophilic acidophilic bacilli［M］．Berlin：Springer Science & Business Media，2008.
[4]	胡贻椿，岳田利，袁亚宏，等．果汁中脂环酸芽孢杆菌（Alicyclobacillus spp.）的危害及其控制［J］．食品科学，2008，29（1）：364-368.
	HU Yi-hun， YUE Tian-li， YUAN Ya-hong，et al ．Hazards and controlling methods of Alicyclobacillus spp．in juice［J］．Food Science，2008，29（1）：364-368.
[5]	蔡瑞，袁亚宏，王周利，等． ε-聚赖氨酸对脂环酸芽孢杆菌生长与产物代谢的影响［J］．农业机械报，2015，46（10）：285-289，327.
	CAI Rui， YUAN Yahong， WANG Zhouli，et al ．Effect of ε-polylysine on growth and guaiacol production of Alicyclobacillus spp.［J］．Transactions of the Chinese Society for Agricultural Machinery，2015，46（10）：285-289，327.
[6]	DURAK M Z， CHUREY J J， DANYLUK M D，et al ．Identification and haplotype distribution of Alicyclobacillus spp．from different juices and beverages［J］．International Journal of Food Microbiology，2010，142（3）：286-291.
[7]	ZHANG J B， YUE T L， YUAN Y H ． Alicyclobacillus contamination in the production line of kiwi products in China［J］．Plos One，2013，8（7）：e67704/1-13.
[8]	王周利，蔡瑞，岳田利，等．果汁中脂环酸芽孢杆菌识别与控制研究进展［J］．农业机械学报，2016，47（10）：221-243.
	WANG Zhouli， CAI Rui， YUE Tianli，et al ．Review of identification and control technology of Alicyclobacillus spp.in fruit juice［J］．Transactions of the Chinese Society for Agricultural Machinery，2016，47（10）：221-243.
[9]	张江波，刘兴华，岳田利，等．陕西猕猴桃汁嗜酸耐热菌污染能力评测与多样性分析［J］．农业机械学报，2015，46（2）：198-207.
	ZHANG Jiangbo， LIU Xinghua， YUE Tianli，et al ．Evaluation of kiwi-fruit juice contaminant capacity of indigenous Alicyclobacillus and its biodiversity［J］．Transactions of the Chinese Society for Agricultural Machinery，2015，46（2）：198-207.
[10]	SETLOW B， COWAN A E， SETLOW P ．Germination of spores of Bacillus subtilis with dodecylamine［J］．Journal of Applied Microbiology，2003，95（3）：637-648.
[11]	EVELYN， FILIPA V M S ．Use of power ultrasound to enhance the thermal inactivation of Clostridium perfringens spores in beef slurry［J］．International Journal of Food Microbiology，2015，206（1）：17-23.
[12]	QIU X， LEE Y T， YUNG P T ．A bacterial spore model of pulsed electric fields on spore morphology change revealed by simulation and SEM［C］∥ Proceedings of 36th Annual International Conference of the IEEE Engineering in Medicine and Biology Society．New York：IEEE，2014：6822-6825.
[13]	LEE S Y， PARK S H， KANG D H ．Inactivation of Alicyclobacillus acidoterrestris spores in apple and orange juice concentrates by gamma irradiation［J］．Journal of Food Protection，2014，77（2）：339-344.
[14]	TORLAK E ．Efficacy of ozone against Alicyclobacillus acidoterrestris spores in apple juice［J］．International Journal of Food Microbiology，2014，172（1）：1-4.
[15]	陈玉飞，帖金凤，蒙倩娜，等．两种过氧化氢消毒液对细菌芽孢杀灭效果的观察［J］．中国消毒学杂志，2019，36（11）：804-807.
	CHEN Yufei， Jinfeng TIE， MENG Qianna，et al ．Observation on the killing effect of two kinds of hydrogen peroxide disinfectants on bacterial spores［J］．Chinese Journal of Disinfection，2019，36（11）：804-807.
[16]	KAWASE K Y F， LUCHESE R H， COELHO G L ．Micronized benzoic acid decreases the concentration necessary to preserve acidic beverages against Alicyclobacillus ［J］．Journal of Applied Microbiology，2013，115（2）：466-474.
[17]	UDADHYAY A， UPADHYAY I， KOLLANOOR-JOHNY A，et al ．Combating pathogenic microorganisms using plant-derived antimicrobials：a minireview of the mechanistic basis［J］．BioMed Research International，2014，2014（55）：761741/1-18.
[18]	马明霞．紫苏酸对酸土脂环酸芽孢杆菌的抑菌机制研究及其保鲜应用［D］．长春：吉林大学，2023.
[19]	SILVA V M F， GIBBS P ． Alicyclobacillus acidoterrestris spores in fruit products and design of pasteurization processes［J］．Trends in Food Science & Technology，2001，12（2）：68-74.
[20]	MIRZAEI-NAJAFGHOLI H， TARIGHI S， GOLMOHAMMADI M，et al ．The effect of citrus essential oils and their constituents on growth of Xanthomonas citri subsp. citri［J］．Molecules，2017，22（4）：E591/1-14.
[21]	TURGIS M，VU KD， DUPONT C，et al ．Combined antimicrobial effect of essential oils and bacteriocins against foodborne pathogens and food spoilage bacteria［J］．Food Research International，2012，48（2）：696-702.
[22]	武思睿．超声、热和Nisin协同处理对枯草杆菌芽孢灭活作用及其在果蔬汁杀菌中的应用［D］．银川：宁夏大学，2023.
[23]	刘锋，陆殷明，陶文静，等．姜黄素与根皮素联合作用对大肠杆菌的抑菌效果［J］．中国食品学报，2023，23（8）：75-83.
	LIU Feng， LU Yinming， TAO Wenjing，et al ．Inhibition of curcumin combined with phloretin against Escherichia coli ［J］．Journal of Chinese Institute of Food Science and Technology，2023，23（8）：75-83.
[24]	王金萧．苹果属植物二氢查尔酮化合物抑菌活性研究［D］．杨凌：西北农林科技大学，2019.
[25]	罗璇．杨梅素与根皮素协同抑菌及其在食品保鲜中的应用研究［D］．武汉：华中农业大学，2022.
[26]	ADIL M， BAIG M H， RUPASINGHE H P V ．Impact of citral and phloretin，alone and in combination，on major virulence traits of Streptococcus pyogenes ［J］．Molecules，2019，24（23）：4237/1-18.
[27]	LI J， ZHAO N， XU R，et al ．Deciphering the antibacterial activity and mechanism of p-coumaric acid against Alicyclobacillus acidoterrestris and its application in apple juice［J］．International Journal of Food Microbiology，2022，378（1）：109822/1-11.
[28]	CAI R， ZHANG M Q，CUI，L，et al ．Antibacterial activity and mechanism of thymol against Alicyclobacillus acidoterrestris vegetative cells and spores［J］．LWT-Food Science & Technology，2019，105（1）：377-384.
[29]	章中，张变飞，尚彬玲，等．高压热杀菌结合葡萄糖对枯草芽孢杆菌芽孢的杀灭效果［J］．食品与生物技术学报，2023，42（5）：88-95.
	ZHANG Zhong， ZHANG Bianfei， SHANG Binling，et al ．Effects of high pressure thermal sterilization combining with glucose on the sterilization of Bacillus subtilis ［J］．Journal of Food Science and Biotechnology，2023，42（5）：88-95.
[30]	FAN L H， HOU F，MUHAMMAD，et al ．Synergistic inactivation and mechanism of thermal and ultrasound treatments against Bacillus subtilis spores［J］．Food Research International，2019，116：1094-1102.
[31]	SAŠA P， ANJA K， LEA D，et al ．Control of Alicyclobacillus spp．vegetative cells and spores in apple juice with rosemary extracts［J］．Food Control，2016，60：205-214.
[32]	MEANEY C A， CARTMAN S T， MCLURE P J，et al ．Optimal spore germination in Clostridium botulinum ATCC 3502 requires the presence of functional copies of SleB and YpeB，but not CwlJ［J］．Anaerobe，2015，34：86-93.
[33]	黎琳莹．异硫氰酸烯丙酯对产气荚膜梭菌的抑制作用及其在肉类食品中的应用［D］．广州：华南理工大学，2023.
[34]	SAHIN O， YONG H E， DRIKS A ．Physical basis tor the adaptive flexibility of Bacillus spore coats［J］．Journal of the Royal Society Interface，2012，9：3156-3160.
[35]	白妍．等离子体活化水对蜡样芽孢杆菌（B .cereus）芽孢的杀灭效果与机理研究［D］．杭州：浙江大学，2019.
[36]	李元政，胡文忠，萨仁高娃，等．天然植物提取物的抑菌机理及其在果蔬保鲜中的应用［J］．食品与发酵工业，2019，45（14）：239-244.
	LI Yuanzheng， HU Wenzhong， SAREN Gaowa，et al ．Antimicrobial mechanisms of natural plant extracts and applications in preserving fruits and vegetables［J］．Food and Fermentation Industries，2019，45（14）：239-244.
[37]	黄筱钧，徐慧，白游震，等．破坏细菌芽孢高抵抗力的研究进展［J］．湖北民族大学学报（医学版），2022，39（4）：48-51.
	HUANG Xiaojun， XU Hui， BAI Youzhen，et al ．Research progress on the high resistance of bacterial spores to destruction［J］．Journal of Hubei University （Medical Edition），2022，39（4）：48-51.