| 2种侵入型乳酸菌生物学特性的研究 |
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| 引用本文： | 曾扬,尹文巧,高可丽,李鼎伟,刘晶,杨玉莹.2种侵入型乳酸菌生物学特性的研究[J].中国畜牧兽医,2021,48(10):3823-3833. |
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| 作者姓名： | 曾扬 尹文巧 高可丽 李鼎伟 刘晶 杨玉莹 |
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| 作者单位： | 长江大学动物科学学院, 荆州 434025 |
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| 基金项目： | 农业农村部畜禽细菌病防治制剂创制重点实验室开放课题（KLPCAAB-2021-04）；国家自然科学基金面上项目（31972646）；长江大学大学生创新训练项目（Yz2020093） |
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| 摘 要： | 本研究旨在选择性能优良的乳酸菌表达载体，为后期研发新型乳酸菌活菌疫苗奠定基础。试验对诱导型和组成型表达的2种侵入型乳酸菌NC8-pSIP409-FnBPA和NC8-pLc23-FnBPA的生物学特性进行研究，从酸耐受性、胆盐耐受性、生长曲线、质粒稳定性、抑菌试验以及药敏试验这6个方面进行鉴定。结果表明，这2种乳酸菌在pH为2.5的酸应激条件下，存活率均达到125%，在质量分数为0.5%的高胆盐溶液中，存活率高达200%。2种类型的乳酸菌均能在9 h时达到平台期，生长性能优良，其中组成型表达菌株NC8-pLc23-FnBPA达到平台期时的D600 nm值为1.3，高于诱导型表达菌株，生长性能优于诱导型菌株。pH生长曲线结果表明，乳酸菌产酸性能无明显性差异，在10 h时，溶液中的pH均降为3.5，产酸能力较强。2种类型乳酸菌的菌体和上清对常见致病菌均有不同程度的抑菌作用，其中菌体的抑菌效果优于上清，且表达FnBPA蛋白的组成型菌体的抑菌效果强于诱导表达型。2种侵入型乳酸菌均对绝大多数的抗生素敏感性高，具有一定的生物安全性。2种类型的乳酸菌的质粒稳定性均>90%。本试验发现诱导型乳酸菌NC8-pSIP409-FnBPA与组成型乳酸菌NC8-pLc23-FnBPA具有相似的生物学特性，其中组成型乳酸菌表达蛋白更方便，无需添加诱导肽便可自主表达，更适合作为新型乳酸菌活菌疫苗的受体菌株，为后期乳酸菌活菌疫苗的研制提供便利。
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| 关 键 词： | 植物乳杆菌 pSIP409载体 诱导型 组成型 生物学特性 |
| 收稿时间： | 2021-04-29 |
Study on Biological Characteristics of Two Invasive Lactic Acid Bacteria |
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| 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 & Veterinary Medicine,2021,48(10):3823-3833. |
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| Authors: | ZENG Yang YIN Wenqiao GAO Keli LI Dingwei LIU Jing YANG Yuying |
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| Institution: | College of Animal Science, Yangtze University, Jingzhou 434025, China |
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| 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 D600 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. |
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| Keywords: | Lactobacillus plantarum pSIP409 vector inducible constitutive biological characteristics |
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