表达牛乳铁蛋白肽的罗伊氏乳杆菌分泌型信号肽的筛选及鉴定

doi:10.11843/j.issn.0366-6964.2025.03.041

Abstract

Abstract:

The aim of this study was to construct a recombinant Lactobacillus reuteri expressing bovine lactoferrin peptides, and to enhance the secretion and expression of lactoferrin peptides by screening different signal peptides, thereby improving the application efficiency of the recombinant strains. A strain of Lactobacillus reuteri isolated in the laboratory was subjected to whole genome sequencing, the signal peptide gene for secreted proteins was screened, and recombinant strains expressing bovine lactoferrin peptides(LFCA)were constructed. The effect of LFCA secreted by the signal peptide was analyzed and compared by Western blot, ELISA, and laser confocal techniques, and the inhibition of proliferative activity of Staphylococcus aureus by LFCA secreted by the recombinant bacteria as well as the half-minimum inhibitory concentration were identified. The results showed that A2, A3, A4 and A8 signal peptides were able to express LFCA, of which A3 and A8 were able to successfully make them secreted into the culture supernatant, and A3 signal peptide increased the secretion and expression level of bovine lactoferrin peptides nearly 2-fold, which did not have significant effect on the growth performance of the strains, and the half-minimum inhibitory concentration of LFCA secreted by the strains was 97.50 μg·mL^-1 against Staphylococcus aureus. This study successfully screened the signal peptide which could secrete lactoferrin peptide efficiently, and the bacteriostatic and antiviral efficiency of the recombinant strain was significantly improved.

Key words: bovine lactoferrin, Lactobacillus reuteri, the recombinant protein secreted signal peptide, antibacterial activity

CLC Number:

S852.69

ZHAO Wenyue, YANG Jing, SHAO Yilan, LI Jiaxuan, JIANG Yanping, CUI Wen, WANG Xiaona, TANG Lijie. Screening and Identification of Secretory Signal Peptide of Lactobacillus reuteri Expressing Lactoferrin Peptide[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(3): 1431-1440.

Figures/Tables 8

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信号肽名称 Name of signal peptide	信号肽序列 Signal peptide sequence
A1	ATGTTGGATCGACACGTTTGGCAAAGGCTTGCTACCGTGGTACTGGCAGCCTTGGTAAT TTTCAGCATCAATACTTTCTCACCAGTA
A2	ATGAAAAAATATATTATCAGTATTAGTAGTATTGTTGCTTTGTTGGTTATAATTCTT GGGTTATCCTTTGTTCCGTGGAAATCGATGGGG
A3	ATGGTGAACGAGATAAATTTCGTTTTTCGACTGCTTAAGCGGATATTTATAATAGGAC TTTTAGTAGGTGGAGGATGGCTTTATTTCAATGATGCACAAGTTCAAGCAACGGCG
A4	ATGCGATTTCGATTCAATAAAAAGACTCAGTATTTACTTTTATCTTTAGTAGCAGTTCT CGGCATTGGTAGTTTTTCC
A5	ATGAAAAAAGAAACAAAAATCATTTGTGGAAGTGTACTTGCATTTTTGATTATTTTAT TTGGGATTGTGGCG
A6	ATGAGTAAAAAATTACTTGCTGGGATATTATTAGGTGGAGTCGCAACATATGCT
A7	ATGGGCAAGGACACTAAAGGCTTCTTTCGCCGGAAATGGTTTTTGCGAATTATTTCGTT AATCCTAGCACTCTTTCTATTCATGTATGTAAATGGG
A8	ATGAAAAAATTTAAGTTGATTAGCGGTTTATTAGTTATGTTCTTAGCATTCTTTATTT TAACGGGATATACA
A9	ATGAAATTTTGGAAGAAAGCACCATTAACAATTGCAGCCTTAACAGTCAGTACCTCTG CAGGAATCACAAGTGTTTCTGCT
A10	ATGCAATCAAGTTTAAAGAAATCTCTTTACTTGGGCCTTGCCGCATTGAGCTTTGCTG GTGTTGCTGCCGTTTCAACGACTGCTTCAGCT

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Screening and Identification of Secretory Signal Peptide of Lactobacillus reuteri Expressing Lactoferrin Peptide

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