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
通讯作者: 党向利
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
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

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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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