乳酸片球菌J1产胞外多糖发酵条件优化

doi:10.11924/j.issn.1000-6850.casb2023-0247

中国农学通报 ›› 2024, Vol. 40 ›› Issue (6): 143-151.doi: 10.11924/j.issn.1000-6850.casb2023-0247

乳酸片球菌J1产胞外多糖发酵条件优化

于文娜¹(), 于连升¹, 王烁¹, 赵丹¹^,², 杜仁鹏¹^,²()

¹ 黑龙江大学，生命科学学院，农业微生物技术教育部工程研究中心，黑龙江省寒区植物基因与生物发酵重点实验室，黑龙江省普通高校微生物重点实验室，哈尔滨 150080
² 河北环境工程学院，河北省农业生态安全重点实验室，河北秦皇岛 066102

收稿日期:2023-03-23 修回日期:2023-06-15 出版日期:2024-02-22 发布日期:2024-02-22
通讯作者:
杜仁鹏，男，1989年出生，黑龙江大兴安岭人，副教授，博士，研究方向：微生物次生代谢产物与生理。通信地址：150080 黑龙江哈尔滨南岗区学府路74号生命科学学院，Tel：0451-86609134，E-mail：durenpeng20096292@163.com。
作者简介:
于文娜，女，2002年出生，山东临沂人，硕士研究生，研究方向：微生物次生代谢产物与生理。通信地址：150080 黑龙江哈尔滨南岗区学府路74号生命科学学院，Tel：0451-86609134，E-mail：yuwenna02@163.com。
基金资助:
黑龙江省自然科学基金优秀青年基金项目“乳酸明串株菌群体感应和第二信使介导右旋糖酐合成的信号通路研究”(YQ2021C030); 中国博士后科学基金资助项目“c-di-GMP和c-di-AMP介导乳酸明串珠菌L2胞外多糖合成的分子调控机制研究”(2022MD713755); 黑龙江省博士后资助项目“柠檬明串珠菌胞外多糖合成途径中信号网络的作用机制研究”(LBH-Z21082); 新时代龙江优秀硕士、博士学位论文项目“肠膜明串珠菌DRP105右旋糖酐生物合成机制的研究”(LJYXL2022-020); 河北省农业生态安全重点实验室开放基金课题项目“乳酸菌胞外多糖的分离纯化及构效关系研究”(2023SYSJJ17); 黑龙江省省属高等学校基本科研业务费科研项目“c-di-AMP调控肠膜明串珠菌DRP105胞外多糖的生物合成机理”(2022-KYYWF-1075)

Exopolysaccharide Production by Pediococcus acidilactici J1: Fermentation Conditions Optimization

YU Wenna¹(), YU Liansheng¹, WANG Shuo¹, ZHAO Dan¹^,², DU Renpeng¹^,²()

¹ Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education/Heilongjiang Provincial Key Laboratory of Plant Genetic Engineering and Biological Fermentation Engineering for Cold Region/Key Laboratory of Microbiology, College of Heilongjiang Province/School of Life Sciences, Heilongjiang University, Harbin 150080
² Hebei University of Environmental Engineering/Hebei Key Laboratory of Agroecological Safety, Qinhuangdao, Hebei 066102

Received:2023-03-23 Revised:2023-06-15 Published-:2024-02-22 Online:2024-02-22

摘要/Abstract

摘要：

乳酸菌(Lactic acid bacteria，LAB)能够合成具有多种功能特性的胞外多糖(Exopolysaccharides，EPS)，其通常具有改善乳制品的风味，抗癌、抗氧化和益生特性等功能。然而LAB EPS的产量较低，限制了EPS的工业化生产应用。为了提高EPS的产量，促进其商品化发展，优化菌株产EPS的发酵条件至关重要。本研究利用单因素试验的方法优化乳酸片球菌(Pediococcus acidilactici)J1发酵产EPS的最佳条件。研究表明：菌株在70 g/L蔗糖、14 g/L牛肉膏、3.5 g/L柠檬酸铵、4 g/L无水乙酸钠、4.00%接种量、培养温度35℃、pH 5.5和160 r/min摇床转速条件下，EPS产量最高，为(72.66±0.02)g/L。本研究通过单因素实验显著提高了P. acidilactici J1的EPS产量，是优化前的4.76倍，为乳酸菌EPS的工业化大规模生产奠定了坚实的理论基础。

关键词: 乳酸片球菌, 胞外多糖, 单因素试验, 发酵条件, 优化

Abstract:

Lactic acid bacteria (LAB) can synthesize exopolysaccharides (EPS) with various functional characteristics, which can improve the flavor of dairy products and have functions such as anticancer, antioxidant, and probiotic properties. However, the low production of LAB EPS limits its industrial production and application. In order to increase the yield of EPS and promote its commercial development, it is very important to optimize the fermentation conditions of strains producing EPS. In this study, the method of single factor test was used to optimize the conditions for EPS production by Pediococcus acidilactici J1. The results showed that under the conditions of 70 g/L sucrose, 14 g/L beef paste, 3.5 g/L ammonium citrate, 4 g/L anhydrous sodium acetate, 4.00% inoculum amount, 35℃ culture temperature, pH 5.5 and 160 r/min shaking speed, EPS yield was the highest, which was (72.66±0.02) g/L. This study significantly improved the EPS yield of P. acidilactici J1, which was 4.76 times of that before optimization. It laid a solid theoretical foundation for the industrial large-scale production of LAB EPS.

Key words: Pediococcus acidilactici, exopolysaccharides, single factor test, fermentation condition, optimization

于文娜, 于连升, 王烁, 赵丹, 杜仁鹏. 乳酸片球菌J1产胞外多糖发酵条件优化[J]. 中国农学通报, 2024, 40(6): 143-151.

YU Wenna, YU Liansheng, WANG Shuo, ZHAO Dan, DU Renpeng. Exopolysaccharide Production by Pediococcus acidilactici J1: Fermentation Conditions Optimization[J]. Chinese Agricultural Science Bulletin, 2024, 40(6): 143-151.

图/表 12

	葡萄糖	麦芽糖	果糖	乳糖	蔗糖
细菌数/(CFU/mL)	1.61E+8	7.03E+7	9.85E+7	4.11E+7	7.69E+8
EPS含量/(g/L)	12.65±0.20	2.58±0.12	11.56±0.15	2.50±0.11	15.27±0.14

表1. 碳源对P. acidilactici J1生长和EPS产量的影响

图1. 蔗糖浓度对P. acidilactici J1生长和EPS产量的影响对EPS含量小于0.05的差异显著性在图中用大写字母表示，对细菌数小于0.05的差异显著性用小写字母表示，下同

	蛋白胨	酵母浸粉	牛肉膏
细菌数/(CFU/mL)	4.77E+8	4.61E+8	1.17E+8
EPS含量/(g/L)	36.15±2.24	43.76±1.07	45.71±1.18

表2. 氮源对P. acidilactici J1生长和EPS产量的影响

图2. 牛肉膏和柠檬酸铵浓度对P. acidilactici J1生长和EPS产量的影响

	磷酸氢二钾	硫酸镁	无水乙酸钠
细菌数/(CFU/mL)	1.01E+8	1.35E+8	1.34E+8
EPS含量/(g/L)	46.540±0.10	51.908±0.07	55.662±0.20

表3. 无机盐对P. acidilactici J1生长和EPS产量的影响

图3. 磷酸氢二钾添加量对P. acidilactici J1生长和EPS产量的影响

图4. 硫酸镁添加量对P. acidilactici J1生长和EPS产量的影响

图5. 无水乙酸钠添加量对P. acidilactici J1生长和EPS产量的影响

图6. 接种量对P. acidilactici J1生长和EPS产量的影响

图7. pH对P. acidilactici J1生长和EPS产量的影响

图8. 温度对P. acidilactici J1生长和EPS产量的影响

图9. 不同的摇床转速对P. acidilactici J1生长和EPS产量的影响

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乳酸片球菌J1产胞外多糖发酵条件优化

Exopolysaccharide Production by Pediococcus acidilactici J1: Fermentation Conditions Optimization

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