The Heterologous Expression and Antibacterial Properties of the Bacteriocin Paracin1.7 in Lactococcus Lactis NZ9000

doi:10.11924/j.issn.1000-6850.casb2021-0463

Abstract

Abstract:

To obtain the bacteriocin with better antibacterial effect, this experiment used heterologous expression method to express the presumed bacteriocin protein 57AA, 62AA, 77AA, 84AA, taking the expression host of Lactococcus lactis NZ9000 and the vector of pNZ8148 to detect the antibacterial effect of heterologous expression products. The results show that the four hypothetical protein 57AA, 62AA, 77AA, and 84AA have antibacterial effects on B. subtilis. Compared with Paracin 1.7 produced by Lactobacillus paracasei HD1.7, the antibacterial activities of 57AA, 62AA, 77AA, and 84AA were improved by 7.57 times, 4.53 times, 5.89 times (very significant difference P<0.01) and 1.20 times (significant difference P<0.05), respectively. In conclusion, the heterologously expressed protein 57AA, 62AA, 77AA, and 84AA, which are putatively expressed as bacteriocins, have better antibacterial effects.

Key words: bacteriocin, Paracin 1.7, Lactococcus lactis, heterologous expression, antibacterial properties

CLC Number:

S182

Pan Yue, Yang Zhixin, Yang Xiaoli, Wang Xu, Song Gang. The Heterologous Expression and Antibacterial Properties of the Bacteriocin Paracin1.7 in Lactococcus Lactis NZ9000[J]. Chinese Agricultural Science Bulletin, 2021, 37(26): 15-23.

Figures/Tables 18

References 25

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序号	名称	用途	来源
1	乳酸乳球菌(Lactococcus lactis)NZ9000	异源表达宿主	黑龙江大学微生物重点实验室保藏
2	副干酪乳杆菌(Lactobacillus paracasei )HD1.7	原初宿主
3	枯草芽胞杆菌(Bacillus subtilis)ATCC 11774	抑菌试验指示菌
4	大肠杆菌(Escherichia coli )DH5α	克隆宿主

序号	名称	用途	来源
1	乳酸乳球菌(Lactococcus lactis)NZ9000	异源表达宿主	黑龙江大学微生物重点实验室保藏
2	副干酪乳杆菌(Lactobacillus paracasei )HD1.7	原初宿主
3	枯草芽胞杆菌(Bacillus subtilis)ATCC 11774	抑菌试验指示菌
4	大肠杆菌(Escherichia coli )DH5α	克隆宿主

序号	名称	特征	大小/bp	用途
1	pNZ8148	Cm ^r携带乳链菌肽诱导型启动子P _nisA	3165	表达载体
2	pMD18-T	—	2692	克隆载体

序号	名称	特征	大小/bp	用途
1	pNZ8148	Cm ^r携带乳链菌肽诱导型启动子P _nisA	3165	表达载体
2	pMD18-T	—	2692	克隆载体

序号	名称	序列(5’-3’)	片段长度	用途
1	57AA-up	GCTCTAGAACGGTGGTTTATGGT	451 bp	扩增全长
1	57AA-down	GGAAGCTTGATGTTGTCCTCCT	451 bp	扩增全长
2	62AA-up	GCCTCTAGATTCGTCGTATTTTCA	423 bp	扩增全长
2	62AA-down	GCAAGCTTATCCTTTCTTCCTGTT	423 bp	扩增全长
3	77AA-up	GCTCTAGACCAAGCAAGGAGATTT	542 bp	扩增全长
3	77AA-down	GCAAGCTTACACCCACTAGGTTTA	542 bp	扩增全长
4	84AA-up	GCGTCTAGATAGGCAGCAAAGAG	516 bp	扩增全长
4	84AA-down	GGAAGCTTCGCCAATAATAACG	516 bp	扩增全长