茶叶抗菌肽粗提物的抑菌活性及其对冷却肉保鲜的影响

doi:10.3969/j.issn.1004-1524.2022.10.21

浙江农业学报 ›› 2022, Vol. 34 ›› Issue (10): 2268-2276.DOI: 10.3969/j.issn.1004-1524.2022.10.21

茶叶抗菌肽粗提物的抑菌活性及其对冷却肉保鲜的影响

王丽芳^a(), 叶良^b, 谢忠稳^c, 党向利^b^,^*()

a. 园艺学院安徽农业大学,安徽合肥 230036
b. 植物保护学院安徽农业大学,安徽合肥 230036
c. 茶树生物学与资源利用国家重点实验室安徽农业大学,安徽合肥 230036

收稿日期:2022-03-02 出版日期:2022-10-25 发布日期:2022-10-26
作者简介:*党向利,E-mail: xldang@ahau.edu.cn
王丽芳(1979—),女,山西大同人,硕士,实验师,主要从事应用微生物与活性成分研究。E-mail: wlf2000@ahau.edu.cn
通讯作者: 党向利
基金资助:
茶树生物学与资源利用国家重点实验室开放基金(SKLTOF20150107);安徽省自然科学基金(2108085MC104)

Antibacterial activity of tea antimicrobial peptide extraction and its effect on preservation of chilled meat

WANG Lifang^a(), YE Liang^b, XIE Zhongwen^c, DANG Xiangli^b^,^*()

a. School of Horticulture, Anhui Agricultural University, Hefei 230036, China
b. School of Plant Protection, Anhui Agricultural University, Hefei 230036, China
c. State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei 230036, China

Received:2022-03-02 Online:2022-10-25 Published:2022-10-26
Contact: DANG Xiangli

摘要/Abstract

摘要：

为了制备茶叶抗菌肽粗提物并将其应用在冷却肉保鲜上,以龙井茶叶为材料提取茶叶抗菌肽粗提物,采用平板抑菌法检测茶叶抗菌肽粗提物的抑菌活性,通过细胞膜渗透率、磷离子泄漏、DNA结合和透射电子显微镜等指标分析茶叶抗菌肽粗提物对细菌的作用机制,通过微生物菌落数量和pH值两个指标评价茶叶抗菌肽粗提物对冷却肉的防腐保鲜效果。结果表明:提取的茶叶抗菌肽粗提物主要包括分子量小于18.4 ku的多肽,其对金黄色葡萄球菌和大肠埃希菌均具有抑菌活性,可造成细菌细胞膜渗透率增强,引起细菌细胞外磷离子浓度增加,造成细菌细胞膜不完整、出现破损,但不与细菌基因组DNA结合。茶叶抗菌肽粗提物可以显著抑制冷却肉中微生物的生长,延缓pH值上升,延长保鲜时间,对冷却肉具有防腐保鲜作用,具有较高的开发利用价值。

关键词: 茶叶, 抗菌肽, 抑菌, 冷却肉, 保鲜

Abstract:

To prepare antimicrobial peptide (AMP) extraction from tea and apply it in chilled meat preservation, tea AMP extraction (TAE) was extracted from Longjing tea. Antibacterial activity of TAE was determined by radial diffusion assay. The mechanism of action of TAE against bacteria was analyzed by membrane permeability, phosphorus ions leakage, DNA binding and transmission electron microscopy. The effect of TAE on preservation of chilled meat was also evaluated based on microbial colonies and pH value. The results showed that TAE mainly contained peptides with a molecular weight of less than 18.4 ku, it showed antibacterial activity against Staphylococcus aureus and Escherichia coli, and it could disrupt bacterial cell membrane and enhance cell membrane permeability, resulting in phosphorus ions leakage. However it did not bind to bacterial genomic DNA. TAE could significantly inhibit the growth of bacteria in chilled meat and delay the increasing of pH value, which could prolong the preservation time and had high development and utilization value in food preservation.

Key words: tea, antimicrobial peptide, antibacterial activity, chilled meat, preservation

中图分类号:

TS201

王丽芳, 叶良, 谢忠稳, 党向利. 茶叶抗菌肽粗提物的抑菌活性及其对冷却肉保鲜的影响[J]. 浙江农业学报, 2022, 34(10): 2268-2276.

WANG Lifang, YE Liang, XIE Zhongwen, DANG Xiangli. Antibacterial activity of tea antimicrobial peptide extraction and its effect on preservation of chilled meat[J]. Acta Agriculturae Zhejiangensis, 2022, 34(10): 2268-2276.

图/表 8

图1 茶叶抗菌肽粗提物SDS-PAGE电泳检测1,10 ku超滤管截留蛋白;2,3 ku超滤管截留蛋白;3,提取的茶叶抗菌肽。

Fig.1 SDS-PAGE of TAE1, Retained proteins by 10 ku ultrafiltration tube; 2, Retained proteins by 3 ku ultrafiltration tube; 3, Extracted TAE.

表1 茶叶抗菌肽粗提物对细菌的抑菌活性

Table 1 Antibacterial activity of TAE against bacteria

处理 Treatment	抑菌圈直径Diameter of inhibition zone/mm
处理 Treatment	金黄色葡萄球菌 Staphylococcus aureus	大肠埃希菌 Escherichia coli
茶叶抗菌肽TAE	13.6±0.44 a	7.9±0.38 a
乳酸链球菌肽Nisin	16.2±0.78 a	4.7±0.42 b

图2 茶叶抗菌肽粗提物对金黄色葡萄球菌细胞膜渗透率的影响

Fig.2 Effect of TAE on cell membrane permeability of S. aureus

图3 茶叶抗菌肽粗提物对金黄色葡萄球菌磷离子泄露的影响

Fig.3 Effect of TAE on phosphate ions leakage of S. aureus

图4 DNA琼脂糖凝胶阻滞分析 M,DNA分子量标准;CK,阴性对照;1,阳性对照;2,茶叶抗菌肽。

Fig.4 Gel retardation analysis of S. aureus genomic DNA M, DNA marker; CK, Negative control; 1, Positive control; 2, TAE.

图5 茶叶抗菌肽粗提物处理对金黄色葡萄球菌形态的影响 A,CK;B和C,茶叶抗菌肽处理。标尺=200 nm。

Fig.5 TEM micrographs of S. aureus treated with TAE A, CK; B and C, TAE teatment. Bar=200 nm.

图6 茶叶抗菌肽处理对冷却肉微生物生长的影响柱上无相同小写字母表示差异显著(P<0.05)。下同。

Fig.6 Effect of TAE on total bacterial count of chilled meat Data on the bars marked without the same lowercase letter indicated significant differences at P<0.05. The same as below.

图7 茶叶抗菌肽处理对冷却肉pH值的影响

Fig.7 Effect of TAE on pH value of chilled meat

参考文献 29

[1]	辛本凯, 王会岩. 抗菌肽在抗菌材料中的研究进展[J]. 吉林医药学院学报, 2021, 42(1): 53-55.
	XIN B K, WANG H Y. Research progress of antimicrobial peptides in antibacterial materials[J]. Journal of Jilin Medical University, 2021, 42(1): 53-55. (in Chinese)
[2]	BROGDEN K A. Antimicrobial peptides: pore formers or metabolic inhibitors in bacteria?[J]. Nature Reviews Microbiology, 2005, 3(3): 238-250. DOI PMID
[3]	BARBOSA PELEGRINI P, DEL SARTO R P, SILVA O N, et al. Antibacterial peptides from plants: what they are and how they probably work[J]. Biochemistry Research International, 2011, 2011: 250349.
[4]	田志环. 抗菌肽的结构与功能[J]. 生物学教学, 2008, 33(7): 4-5.
	TIAN Z H. Structure and function of antimicrobial peptides[J]. Biology Teaching, 2008, 33(7): 4-5. (in Chinese)
[5]	THEVISSEN K, KRISTENSEN H H, THOMMA B P H J, et al. Therapeutic potential of antifungal plant and insect defensins[J]. Drug Discovery Today, 2007, 12(21/22): 966-971. DOI URL
[6]	HEGEDÜS N, MARX F. Antifungal proteins: more than antimicrobials?[J]. Fungal Biology Reviews, 2013, 26(4): 132-145. PMID
[7]	周晓馥, 苗璐, 高峰, 等. 利用生物信息学对植物抗菌肽的预测与分析[J]. 生物技术, 2014, 24(3): 91-95.
	ZHOU X F, MIAO L, GAO F, et al. Bioinformatics forecast and analysis of plant antimicrobial peptides[J]. Biotechnology, 2014, 24(3): 91-95. (in Chinese with English abstract)
[8]	NAWROT R, BARYLSKI J, NOWICKI G, et al. Plant antimicrobial peptides[J]. Folia Microbiologica, 2014, 59(3): 181-196. DOI PMID
[9]	魏志文. 绿茶中四种儿茶素单体(EC、EGC、ECG、EGCG)和槲皮素单体分离制备[D]. 合肥: 安徽农业大学, 2009.
	WEI Z W. Separation and preparation of four individual catechins (EC, EGC, ECG, EGCG) and qurecetin monomer from green tea[D]. Hefei: Anhui Agricultural University, 2009. (in Chinese with English abstract)
[10]	MO H Z, ZHU Y, CHEN Z M. Microbial fermented tea: a potential source of natural food preservatives[J]. Trends in Food Science & Technology, 2008, 19(3): 124-130.
[11]	PERUMALLA A V S, HETTIARACHCHY N S. Green tea and grape seed extracts: potential applications in food safety and quality[J]. Food Research International, 2011, 44(4): 827-839. DOI URL
[12]	王永红, 张淑蓉. 冷鲜肉的保鲜技术研究进展[J]. 粮油食品科技, 2012, 20(1): 48-51.
	WANG Y H, ZHANG S R. Research progress on keeping chilled meat fresh[J]. Science and Technology of Cereals, Oils and Foods, 2012, 20(1): 48-51. (in Chinese with English abstract)
[13]	李新福, 张威, 李超, 等. 不同保鲜剂对冷鲜猪肉品质的影响[J]. 安徽农业科学, 2020, 48(13): 183-188.
	LI X F, ZHANG W, LI C, et al. Effects of different preservatives on storage quality of cold fresh pork[J]. Journal of Anhui Agricultural Sciences, 2020, 48(13): 183-188. (in Chinese)
[14]	XIE Y C, ZHANG Y, XIE Y K, et al. Radio frequency treatment accelerates drying rates and improves vigor of corn seeds[J]. Food Chemistry, 2020, 319: 126597. DOI URL
[15]	LU M Q, HAN J Y, ZHU B Y, et al. Significantly increased amino acid accumulation in a novel albino branch of the tea plant (Camellia sinensis)[J]. Planta, 2019, 249(2): 363-376. DOI PMID
[16]	DANG X L, TIAN J H, YANG W Y, et al. Bactrocerin-1: a novel inducible antimicrobial peptide from pupae of oriental fruit fly Bactrocera dorsalis Hendel[J]. Archives of Insect Biochemistry and Physiology, 2009, 71(3): 117-129. DOI URL
[17]	DANG X L, ZHENG X X, WANG Y S, et al. Antimicrobial peptides from the edible insect Musca domestica and their preservation effect on chilled pork[J]. Journal of Food Processing and Preservation, 2020, 44(3): e14369.
[18]	KLUBTHAWEE N, ADISAKWATTANA P, HANPITHAKPONG W, et al. A novel, rationally designed, hybrid antimicrobial peptide, inspired by cathelicidin and aurein, exhibits membrane-active mechanisms against Pseudomonas aeruginosa[J]. Scientific Reports, 2020, 10: 9117. DOI URL
[19]	段静芸, 徐幸莲, 周光宏. 壳聚糖在冷却鲜猪肉保鲜中的应用研究[J]. 食品工业科技, 2001, 22(4): 26-28.
	DUAN J Y, XU X L, ZHOU G H. Study on the application of chitosan in preservation of chilled fresh pork[J]. Science and Technology of Food Industry, 2001, 22(4): 26-28. (in Chinese with English abstract)
[20]	MAHLAPUU M, BJÖRN C, EKBLOM J. Antimicrobial peptides as therapeutic agents: opportunities and challenges[J]. Critical Reviews in Biotechnology, 2020, 40(7): 978-992. DOI PMID
[21]	MAHLAPUU M, HÅKANSSON J, RINGSTAD L, et al. Antimicrobial peptides: an emerging category of therapeutic agents[J]. Frontiers in Cellular and Infection Microbiology, 2016, 6: 194. DOI PMID
[22]	刘唤明, 孙力军, 王雅玲, 等. 纳豆菌脂肽对金黄色葡萄球菌抑菌机理的研究[J]. 食品工业科技, 2012, 33(11): 109-112.
	LIU H M, SUN L J, WANG Y L, et al. Study on antibacterial mechanism of lipopeptide from Bacillus natto against Staphyloccocus aureus[J]. Science and Technology of Food Industry, 2012, 33(11): 109-112. (in Chinese with English abstract)
[23]	YANG S, LI J, AWEYA J J, et al. Antimicrobial mechanism of Larimichthys crocea whey acidic protein-derived peptide (LCWAP) against Staphylococcus aureus and its application in milk[J]. International Journal of Food Microbiology, 2020, 335: 108891. DOI URL
[24]	陈飞龙, 刘渔珠, 彭勃, 等. 抗菌肽F1对金黄色葡萄球菌的胞内作用机制[J]. 食品科学, 2017, 38(6): 36-41.
	CHEN F L, LIU Y Z, PENG B, et al. Intracellular mechanism of action of antimicrobial peptide F1 on Staphylococcus aureus[J]. Food Science, 2017, 38(6): 36-41. (in Chinese with English abstract)
[25]	苏冠芳, 郝刚, 李莉蓉, 等. 抗菌肽buforinⅡ衍生物抑制细菌核酸合成的机制研究[J]. 中国抗生素杂志, 2012, 37(3): 190-195.
	SU G F, HAO G, LI L R, et al. Antibacterial peptides buforinⅡ-analogues on bacteria by inhibition of DNA synthesis[J]. Chinese Journal of Antibiotics, 2012, 37(3): 190-195. (in Chinese with English abstract)
[26]	CARDOSO M H, MENEGUETTI B T, COSTA B O, et al. Nonlytic antibacterial peptides that translocate through bacterial membranes to act on intracellular targets[J]. International Journal of Molecular Sciences, 2019, 20(19): 4877. DOI URL
[27]	程述震, 王晓拓, 王志东. 冷鲜肉保鲜技术研究进展[J]. 食品研究与开发, 2017, 38(16): 194-198.
	CHENG S Z, WANG X T, WANG Z D. Research progress on preservation methods for chilled meat[J]. Food Research and Development, 2017, 38(16): 194-198. (in Chinese with English abstract)
[28]	李柯欣. 茶多酚的提取、抑菌作用与抑菌机理研究[D]. 成都: 西华大学, 2017.
	LI K X. Study on extraction, bacteriostasis and bacteriostatic mechanism of tea polyphenols[D]. Chengdu: Xihua University, 2017. (in Chinese with English abstract)
[29]	JAY J M. Modern food microbiology[M]. 6th ed. Gaithersburg, MD: Aspen Publishers, 2000.

茶叶抗菌肽粗提物的抑菌活性及其对冷却肉保鲜的影响

Antibacterial activity of tea antimicrobial peptide extraction and its effect on preservation of chilled meat

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