Deng Mengyin, Wu Quanlong, Yang Na, Teng Da, Wang Yexuan, Hao Ya, Lu Haiqiang, Mao Ruoyu, Wang Jianhua
Gene Engineering Laboratory, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
Innovative Team of Antimicrobial Peptides and Alternatives to Antibiotics, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
Int J Mol Sci. 2024 Dec 5;25(23):13095. doi: 10.3390/ijms252313095.
To address the limited supply of serum albumin (SA) and potential pathogen contamination, focus has been concentrated on the heterologous expression of human serum albumin (HSA), particularly in . However, there are rare studies on feline serum albumin (FSA), which requires a large amount in pet foods and clinical treatment. In this work, the codon-optimized recombinant feline serum albumin (rFSA) sequence was designed and transferred into GS115 for recombinant expression. The high-level expression strain was selected by a high concentration of G418, followed by plate and shaker screening. At the 5 L fermenter level, the total protein concentration reached 3.89 mg/mL after 113 h of induction. At lower concentrations (1-4 μM), rFSA exhibited a potent free radical scavenging capacity, reaching 99% and 60% for ABTS• and •O, respectively, which surpassed that of natural plasma-derived FSA. The secondary structure and stability of rFSA were found to be consistent with those of FSA. Additionally, an in vivo safety assay in mice showed no significant difference between the rFSA group and the normal saline group in terms of body weight changes, complete blood count, serum biochemistry, inflammatory factors, and tissue sections. These results above indicate that is the optimal host for the high preparation of rFSA. Furthermore, rFSA has been demonstrated to be relatively safe, which paves the way for subsequent industrialized production and its application in pet foods and veterinary clinics.
为了解决血清白蛋白（SA）供应有限以及潜在病原体污染的问题，研究重点一直集中在人血清白蛋白（HSA）的异源表达上，尤其是在[此处原文缺失具体内容]。然而，关于猫血清白蛋白（FSA）的研究很少，而FSA在宠物食品和临床治疗中需求量很大。在这项工作中，设计了密码子优化的重组猫血清白蛋白（rFSA）序列，并将其转入GS115进行重组表达。通过高浓度的G418筛选出高表达菌株，随后进行平板和摇瓶筛选。在5 L发酵罐水平，诱导113 h后总蛋白浓度达到3.89 mg/mL。在较低浓度（1 - 4 μM）下，rFSA表现出强大的自由基清除能力，对ABTS•和•O的清除率分别达到99%和60%，超过了天然血浆来源的FSA。发现rFSA的二级结构和稳定性与FSA一致。此外，在小鼠体内进行的安全性试验表明，rFSA组与生理盐水组在体重变化、全血细胞计数、血清生化、炎症因子和组织切片方面没有显著差异。上述结果表明[此处原文缺失具体内容]是高制备rFSA的最佳宿主。此外，rFSA已被证明相对安全，这为后续的工业化生产及其在宠物食品和兽医诊所的应用铺平了道路。
