2种侵入型乳酸菌生物学特性的研究

doi:10.16431/j.cnki.1671-7236.2021.10.035

Abstract

Abstract: The purpose of this research was to select the expression vector of lactic acid bacteria (LAB) with excellent performance, and lay the foundation for the new live vaccine of LAB in the later period. The biological characteristics of the two types invasive LAB NC8-pSIP409-FnBPA and NC8-pLc23-FnBPA with constitutive and inducible expression were researched from acid tolerance, bile salt tolerance, growth curve, plasmid stability, bacteriostasis test and drug susceptibility test. The results showed that the survival rate of the two strains under the acid stress condition of pH 2.5 reached 125%, and the survival rate was as high as 200% in the high bile salt solution with a mass fraction of 0.5%. The plateau of the two types strains were reached at 9 h that denoted excellent growth performance. The D_{600 nm} value of the constitutively expressed strain NC8-pLc23-FnBPA was 1.3 when reached it's plateau, which was higher than that of the inducible expression strain and showed better growth performance. The growth curve of pH results showed that there was no significant difference in the acid production performance of LAB and the pH in the solution dropped to 3.5 at 10 h which indicated the acid production capacity was strong. The bacterial cells and supernatants of the two types of strains had different degrees of antibacterial effect on common pathogenic bacteria. Among them, the antibacterial effect of the bacteria was better than that of the supernatant, and the antibacterial effect of the constitutive bacteria expressing FnBPA protein was stronger than the other one. Both of the invasive lactic acid bacteria were highly sensitive to most antibiotics and had certain biological safety. The plasmid stability of the two strains were relatively strong even greater than 90%. This study found that the inducible LAB NC8-pSIP409-FnBPA strain and the constitutive NC8-pLc23-FnBPA strain had similar biological characteristics. Among them, it was more convenient for the constitutive LAB to express protein even without adding inducing peptides, which was more suitable to be the recipient for a new LAB strain of live vaccines and provided convenience for the later development of live LAB vaccines.

Key words: Lactobacillus plantarum; pSIP409 vector; inducible; constitutive; biological characteristics

CLC Number:

S852.5

ZENG Yang, YIN Wenqiao, GAO Keli, LI Dingwei, LIU Jing, YANG Yuying. Study on Biological Characteristics of Two Invasive Lactic Acid Bacteria[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(10): 3823-3833.

References

[1] 刘晶, 杨桂连, 王春凤, 等.乳酸菌作为DNA疫苗载体的研究进展[J]. 中国兽医杂志, 2016, 52(7):67-68. LIU J, YANG G L, WANG C F, et al.Research progress of lactic acid bacteria as DNA vaccine carriers[J]. Chinese Journal of Veterinary Medicine, 2016, 52(7):67-68.(in Chinese)
[2] LUIS G B, CAMILLE A, JEAN-PAUL M, et al.Engineering Lactococci and Lactobacilli for human health[J]. Current Opinion in Microbiology, 2013, 16(3):278-283.
[3] YUEH-TING T, PO-CHING C, TZU-MING P.The immunomodulatory effects of lactic acid bacteria for improving immune functions and benefits[J]. Applied Microbiology and Biotechnology, 2012, 96(4):853-862.
[4] DE V M, SCHREZENMEIR J.Probiotics, prebiotics, and synbiotics[J]. Advances in Biochemical Engineering/Biotechnology, 2008, 111:1-66.
[5] JULIANA F A, NATALIA M B, MILOUD M, et al.Expression of fibronectin binding protein A (FnBPA) from Staphylococcus aureus at the cell surface of Lactococcus lactis improves its immunomodulatory properties when used as protein delivery vector[J]. Vaccine, 2016, 34(10):1312-1318.
[6] LIU Q, JIANG Y L, YANG W T, et al.Protective effects of a food-grade recombinant Lactobacillus plantarum with surface displayed AMA1 and EtMIC2 proteins of Eimeria tenella in broiler chicken[J]. Microbial Cell Factories, 2020, 19(1):28.
[7] 刘乃芝, 亓秀晔, 郭杨丽, 等.基因工程重组乳酸菌防控动物疫病的研究进展[J]. 中国畜牧兽医, 2020, 47(4):1199-1208. LIU N Z, QI X Y, GUO Y L, et al.Research progress of genetic engineering recombinant lactic acid bacteria on epidemic prevention and control of animal diseases[J]. China Animal Husbandry & Veterinary Medicine, 2020, 47(4):1199-1208.(in Chinese)
[8] LIU Y Y, YANG W T, SHI S H, et al.Immunogenicity of recombinant Lactobacillus plantarum NC8 expressing goose parvovirus VP2 gene in BALB/c mice[J]. Journal of Veterinary Science, 2017, 18(2):159-167.
[9] 姜延龙, 王春凤, 刘晶.一种基于FnBPA蛋白的侵入型乳酸菌:CN106544352A[P].2017-03-29. JIANG Y L, WANG C F, LIU J.An invasive lactic acid bacteria based on FnBPA protein:CN106544352A[P].2017-03-29.(in Chinese)
[10] LIU J, YANG G, GAO X, et al.Recombinant invasive Lactobacillus plantarum expressing fibronectin binding protein A induce specific humoral immune response by stimulating differentiation of dendritic cells[J]. Beneficial Microbes, 2019, 10(5):589-604.
[11] LIU Y, JIANG Y L, LIU J, et al.Recombinant invasive Lactobacillus plantarum expressing the Eimeria tenella fusion gene TA4 and AMA1 induces protection against coccidiosis in chickens[J]. Veterinary Parasitology, 2020, 283:109161.
[12] 王宜, 方小伟, 刘慧娟, 等.单核细胞增生李斯特菌SigB调控GadD3的抗酸应激作用[J]. 动物医学进展, 2020, 41(1):45-49. WANG Y, FANG X W, LIU H J.SigB regulated GadD3 in acid resistance of Listeria monocytogenes[J]. Progress in Veterinary Medicine, 2020, 41(1):45-49.(in Chinese)
[13] 张赞, 刘晶, 高兴, 等.表面展示柔嫩艾美耳球虫EtMIC2蛋白的侵入型乳酸菌的免疫特性研究[J]. 中国兽医科学, 2019, 49(5):619-624. ZHANG Z, LIU J, GAO X, et al.Immune properties of invasive lactic acid bacteria expressing EtMIC2 protein of Eimeria tenella on its surface[J]. Veterinary Science in China, 2019, 49(5):619-624.(in Chinese)
[14] WANG M, GAO Z Q, ZHANG Y G, et al.Lactic acid bacteria as mucosal delivery vehicles:A realistic therapeutic option[J]. Applied Microbiology and Biotechnology, 2016, 100(13):5691-5701.
[15] 崔月倩, 王菁蕊, 王艳萍.乳酸菌基因表达载体及其应用研究进展[J]. 食品科学, 2015, 36(9):224-229. CUI Y Q, WANG J R, WANG Y P.A review of research on lactic acid bacteria vectors for gene expression and their applications[J]. Food Science, 2015, 36(9):224-229.(in Chinese)
[16] AGNIESZKA W, PATRYCJA K, JACEK B, et al.Lactic acid bacteria-20 years exploring their potential as live vectors for mucosal vaccination[J]. Applied Microbiology and Biotechnology, 2015, 99(7):2967-2977.
[17] 匡珍, 李学英, 徐春霞, 等.乳酸菌细菌素研究进展及其在水产养殖和加工中的应用[J]. 食品工业科技, 2019, 40(4):292-298. KUANG Z, LI X Y, XU C X, et al.Research progress of bacteriocins from lactic acid bacteria and its application in aqua culture and processing[J]. Science and Technology of Food Industry, 2019, 40(4):292-298.(in Chinese)
[18] 刘俊生.植物乳杆菌对沙门氏菌感染预防作用的研究[D].无锡:江南大学, 2019. LIU J S.Study on the preventive effects of Lactobacillus plantarum against Salmonella infection[D].Wuxi:Jiangnan University, 2019.(in Chinese)
[19] CAI R P, JIANG Y L, YANG W, et al.Surface-displayed IL-10 by recombinant Lactobacillus plantarum reduces Th1 responses of RAW264.7 Cells stimulated with Poly(I:C) or LPS[J]. Journal of Microbiology and Biotechnology, 2016, 26(2):421-431.

Study on Biological Characteristics of Two Invasive Lactic Acid Bacteria

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