High‐efficiency transformation of Streptococcus thermophilus using electroporation
High‐efficiency transformation of Streptococcus thermophilus using electroporation
复制标题
利用电穿孔高效转化嗜热链球菌
DOI：
10.1002/jsfa.11292
复制
发表时间：
2021
影响因子：
4.1
通讯作者：
Lian‐Zhong Ai
中科院分区：
文献类型：
--
作者：
Ling‐Hui Kong;Zhi‐Qiang Xiong;Yong‐Jun Xia;Lian‐Zhong Ai
背景嗜热链球菌是一种重要的乳酸菌，广泛应用于食品发酵，有利于提高食品品质。然而，目前的遗传转化系统对s是低效的。这阻碍了它的进一步研究。结果建立了3种s的电穿孔转化方法。结果表明，该菌株的转化效率可达1.3 × 106CFU/μg DNA，是未优化菌株的32倍。此外，转录分析表明，一系列胜任基因inS。优化后，thermophilus‐3的表达再次显著上调，表明转化效率的提高与能力基因的表达水平有关。此外，为了证明它们的潜力，通过表达能力基因（comEA， cbpdd和comx）来提高转化效率。过表达comea获得了最大的转化效率，比对照高14倍。结论本研究首次报道了增强可转化性蛋白的能力基因表达。嗜热菌，这对理想特性菌株的发展有积极的影响。©2021化学工业协会。
BACKGROUNDStreptococcus thermophilus, one of the most important lactic acid bacteria, is widely used in food fermentation, which is beneficial to improve food quality. However, the current genetic transformation systems are inefficient forS. thermophilusS‐3, which hinders its further study.RESULTSWe developed three electroporation transformation methods forS. thermophilusS‐3, and optimized various parameters to enhance the transformation efficiency up to 1.3 × 106CFU/μg DNA, which was 32‐fold higher than that of unoptimized. Additionally, transcriptional analysis showed that a series of competence genes inS. thermophilusS‐3 were remarkedly up‐regulated after optimization, indicating that improvement of transformation efficiency was attributed to the expression level of competence genes. Furthermore, to prove their potential, expression of competence genes (comEA,cbpDandcomX) were employed to increase transformation efficiency. The maximum transformation efficiency was obtained by overexpression ofcomEA, which was 14‐fold higher than that of control.CONCLUSIONThis is the first report of competence gene expression for enhancing transformability inS. thermophilus, which exerts a positive effect on the development of desirable characteristics strains. © 2021 Society of Chemical Industry.