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通过共表达磷酸果糖激酶和葡萄糖激酶在耐头孢菌素乳酸杆菌中实现高产量乳酸用于抗菌特性研究

Achieving High Yield of Lactic Acid for Antimicrobial Characterization in Cephalosporin-Resistant Lactobacillus by the Co-Expression of the Phosphofructokinase and Glucokinase.

作者信息

Gong Yahui, Li Tiyuan, Li Shiyu, Jiang Zhenyou, Yang Yan, Huang Junli, Liu Zhaobing, Sun Hanxiao

机构信息

Institute of Genomic Medicine, College of Pharmacy, Jinan University, Guangzhou 510632, P.R. China.

Clinical Medical College of Jinan University, Guangzhou 510632, P.R. China.

出版信息

J Microbiol Biotechnol. 2016 Jun 28;26(6):1148-61. doi: 10.4014/jmb.1601.01043.

Abstract

Lactobacilli are universally recognized as probiotics that are widely used in the adjuvant treatment of inflammatory diseases, such as vaginitis and enteritis. With the overuse of antibiotics in recent years, the lactobacilli in the human body are killed, which could disrupt the microecological balance in the human body and affect health adversely. In this work, cephalosporin-resistant Lactobacillus casei RL20 was obtained successfully from the feces of healthy volunteers, which possessed a stable genetic set. However, the shortage of lactic acid (72.0 g/l at 48 h) by fermentation did not meet the requirement for its use in medicine. To increase the production of lactic acid, the functional genes pfk and glk were introduced into the wild strain. A yield of 144.2 g/l lactic acid was obtained in the transgenic L. casei RL20-2 after fermentation for 48 h in 1 L of basic fermentation medium with an initial glucose concentration of 100 g/l and increasing antibacterial activity. These data suggested that L. casei RL20-2 that exhibited a high yield of lactic acid may be a potential probiotic to inhibit the spread of bacterial infectious diseases and may be used for vaginitis therapy.

摘要

乳酸杆菌被公认为益生菌,广泛用于阴道炎和肠炎等炎症性疾病的辅助治疗。近年来,随着抗生素的过度使用,人体中的乳酸杆菌被杀死,这可能会破坏人体的微生态平衡并对健康产生不利影响。在这项工作中,成功从健康志愿者的粪便中获得了对头孢菌素耐药的干酪乳杆菌RL20,其具有稳定的基因组。然而,发酵产生的乳酸短缺(48小时时为72.0 g/l)不符合其在医学上使用的要求。为了提高乳酸产量,将功能基因pfk和glk导入野生菌株。在初始葡萄糖浓度为100 g/l的1 L基础发酵培养基中发酵48小时后,转基因干酪乳杆菌RL20-2的乳酸产量达到144.2 g/l,且抗菌活性增强。这些数据表明,表现出高乳酸产量的干酪乳杆菌RL20-2可能是抑制细菌传染病传播的潜在益生菌,可用于阴道炎治疗。

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