[1]程世前,康竞艺,陈萌轩,等.猪传染性胸膜肺炎疫苗抗原ApxⅡ在谷氨酸棒杆菌中的表达及发酵条件优化[J].江苏农业学报,2024,(12):2310-2316.[doi:doi:10.3969/j.issn.1000-4440.2024.12.014]
　CHENG Shiqian,KANG Jingyi,CHEN Mengxuan,et al.Expression of porcine contagious pleuropneumonia vaccine antigen ApxⅡ in Corynebacterium glutamicum and optimization of fermentation conditions[J].,2024,(12):2310-2316.[doi:doi:10.3969/j.issn.1000-4440.2024.12.014]
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猪传染性胸膜肺炎疫苗抗原ApxⅡ在谷氨酸棒杆菌中的表达及发酵条件优化()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2024年12期
页码:
2310-2316
栏目:
畜牧兽医·水产养殖·益虫饲养
出版日期:
2024-12-30

文章信息/Info

Title:
Expression of porcine contagious pleuropneumonia vaccine antigen ApxⅡ in Corynebacterium glutamicum and optimization of fermentation conditions
作者:
程世前1; 康竞艺1; 陈萌轩1; 刘秀霞1; 2; 3; 杨艳坤1; 2; 3; 白仲虎1; 2; 3
(1.江南大学粮食发酵与食品生物制造国家工程研究中心,江苏无锡214122；2.江南大学工业生物技术教育部重点实验室,江苏无锡214122；3.江苏省生物活性制品加工工程技术研究中心,江苏无锡214122)
Author(s):
CHENG Shiqian1; KANG Jingyi1; CHEN Mengxuan1; LIU Xiuxia1; 2; 3; YANG Yankun1; 2; 3; BAI Zhonghu1; 2; 3
(1.National Engineering Research Center of Cereal Fermentation and Food Biomanufacturing, Jiangnan University, Wuxi 214122, China;2.Key Laboratory of Industrial Biotechnongy, Ministry of Education, Jiangnan University, Wuxi 214122, China;3.Jiangsu Engineering and Technology Research Centre for Bioactive Product Processing, Wuxi 214122, China)
关键词:
细胞毒素ApxⅡ; 谷氨酸棒杆菌; 表达
Keywords:
cytotoxin ApxⅡ; Corynebacterium glutamicum; expression
分类号:
S858.28
DOI:
doi:10.3969/j.issn.1000-4440.2024.12.014
文献标志码:
A
摘要:
猪胸膜肺炎放线杆菌是一种高度传染性、致死性呼吸道疾病,已造成养猪业严重损失。细胞毒素Apx Ⅱ是猪胸膜肺炎PCP(Porcine contagious pleuropneumonia)的主要抗原之一,可以作为猪胸膜肺炎疫苗的有效成分。谷氨酸棒杆菌(Corynebacterium glutamicum)具有无内毒素和较发达分泌系统的优点,是非常有潜力的重组细菌疫苗蛋白质表达系统。利用C.glutamicum表达系统生产Apx Ⅱ,实现其高效稳定表达,可为疫苗生产提供技术基础。前期研究已构建Apx Ⅱ分泌表达菌株,为进一步提高Apx Ⅱ的表达产量,通过在24孔板发酵优化,得到最优的培养基为CGXⅡ-YT,最优的表达温度为30 ℃,IPTG浓度为1.0 mmol/L,加入IPTG前培养6 h,发酵培养时间为24 h。进一步在摇瓶中放大发酵培养结果与24孔板发酵培养结果相同。最后在容积为5 L发酵罐中对Apx Ⅱ进行表达,与使用基础发酵培养基发酵相比，使用CGXⅡ-YT发酵培养基，且在优化后的发酵条件下Apx Ⅱ表达量明显提高,达到了114.60 mg/L。本研究实现了Apx Ⅱ在谷氨酸棒杆菌中的分泌表达,为其进一步扩大化生产提供了基础。
Abstract:
Actinobacillus pleuropneumoniae is a highly infectious and fatal respiratory disease, which has caused serious losses in the pig industry. Apx Ⅱ is one of the main antigens of porcine contagious pleuropneumonia (PCP), which can be used as an effective component of porcine pleuropneumonia vaccine. Corynebacterium glutamicum has the advantages of endotoxin-free and more developed secretion system, and is a very promising recombinant bacterial vaccine protein expression system. The expression system of C. glutamicum was used to produce Apx Ⅱ and achieve its efficient and stable expression, which could provide a research basis for vaccine production. Apx Ⅱ secretory expression strains were constructed in previous studies. In order to further improve the expression yield of Apx Ⅱ, the fermentation in 24-well plates was optimized. The optimal medium was CGXII-YT, and the optimal expression temperature was 30 ℃. The concentration of IPTG was 1.0 mmol/L, the culture time was 6 h before adding IPTG, and the fermentation culture time was 24 h. The results were the same as those of 24-well plate fermentation. Finally, Apx Ⅱ was expressed in a 5 L fermentor. Compared with the basic fermentation medium, the Apx Ⅱ expression yield was significantly increased by using CGXⅡ-YT fermentation medium, and the Apx Ⅱ expression yield reached 114.60 mg/L under the optimized fermentation conditions. In this study, the secretory expression of Apx Ⅱ in Corynebacterium glutamicum was realized, which provided a basis for its further expansion of production.

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备注/Memo

备注/Memo:
收稿日期：2024-02-08基金项目：国家自然科学基金项目(22078128)作者简介：程世前(1996-),男,河南义马人,硕士,主要从事发酵工程、医药蛋白表达等方面的研究。(E-mail)c5886602123@163.com通讯作者：白仲虎,(E-mail)baizhonghu@jiangnan.edu.cn
更新日期/Last Update: 2025-01-23