Abstract:
BACKGROUND: Human serum albumin (HSA) is the most abundant protein in plasma, playing crucial roles in regulating osmotic pressure and maintaining protein homeostasis. It is widely applied in the clinical treatment of various diseases. HSA can be purified from plasma or produced using recombinant DNA technology. Due to the improved efficiency and reduced costs, a growing body of research has focused on enhancing albumin production through bacterial strain overexpression. However, there have been few studies on the effect of albumin on the characteristics of the overexpressing-strain itself, particularly stress resistance. In this study, we utilized Lactiplantibacillus plantarum (L. plantarum) AR113 as the expression host and successfully constructed the albumin overexpression strain AR113-pLLY01 through gene editing technology. The successful expression of albumin was achieved and subsequently compared with the wild-type strain AR113-pIB184.
RESULTS: The results demonstrated that the survival rate of AR113-pLLY01 was also significantly better than that of AR113-pIB184 after lyophilization. In addition, AR113-pLLY01 exhibited a significantly better protective effect than AR113-pIB184 at pH 3, indicating that albumin possesses a certain tolerance to acidic stress. At bile salt concentrations higher than 0.03%, both strains showed limited growth, but at a concentration of 0.02%, AR113-pLLY01 had a significant protective effect.
CONCLUSIONS: This study suggest that albumin can improve strain tolerance, which has significant implications for future applications. © 2024 Society of Chemical Industry.
摘要:
背景:人血清白蛋白(HSA)是血浆中含量最高的蛋白质,在调节渗透压和维持蛋白质稳态中起着至关重要的作用。广泛应用于临床各种疾病的治疗。HSA可以从血浆中纯化或使用重组DNA技术产生。由于提高效率和降低成本,越来越多的研究集中在通过细菌菌株的过度表达来增强白蛋白的产生。然而,关于白蛋白对过度表达菌株本身特性的影响的研究很少,特别是抗压力。在这项研究中,我们利用了植物乳杆菌(L.plantarum)AR113为表达宿主,通过基因编辑技术成功构建了白蛋白过表达菌株AR113-pLLY01。实现了白蛋白的成功表达,随后与野生型菌株AR113-pIB184进行比较。
结果:结果表明,冻干后AR113-pLLY01的存活率也明显优于AR113-pIB184。此外,AR113-pLLY01在pH3时表现出比AR113-pIB184明显更好的保护作用,表明白蛋白对酸性胁迫具有一定的耐受性。在胆汁盐浓度高于0.03%时,两种菌株都表现出有限的生长,但浓度为0.02%,AR113-pLLY01具有显著的保护作用。
结论:这项研究表明,白蛋白可以提高菌株的耐受性,这对未来的应用具有重要意义。©2024化学工业学会。