Félix Mellanie K C, Deusdará Tullio T, Brito Hélio S, Santos Gil R, Leite Eduardo R T, Chapla Vanessa M, Viana Kelvinson F, Brandi Igor V, de Almeida Maria Edilene M, Mariúba Luis André M, Nogueira Paulo A, da Silva Elizângela F, Glória Juliane C, da Silva Raquel Stefanni R, Braga Darleide Dos S, de Lima Anderson M, Soares Andreimar M, Cangussu Alex Sander R
Graduate Program for in Biodiversity and Biotechnology of Legal Amazon, Federal University of Tocantins, Palmas 77001-090, TO, Brazil.
Bioprocess Engineering and Biotechnology, Federal University of Tocantins, Gurupi 77410-530, TO, Brazil.
Microorganisms. 2025 Jun 26;13(7):1481. doi: 10.3390/microorganisms13071481.
is a common pathogen in domestic animals and humans, and alpha-toxin is the main cause of its pathogenesis. Because it is a fastidious organism, obtaining alpha-toxin is expensive. Therefore, we proposed an in silico study to synthesize epitopes in cultures of Escherichia coli BL21 pLysS (DE3). First, we used a stirred-tank bioreactor, developing a dry mass yield (DMY) of 0.77 g/L in batch cultures and 1.03 g/L in fed-batch cultures, without acetic acid production. With scale-up using a system without mechanical agitation, there was a higher DMY (1.20 g/L) with 0.56 mmol/mL of alpha-toxin epitope 1 (DE3/Ep1) and 0.61 mmol/mL of alpha-toxin epitope 2 (DE3/Ep2), with a similar profile for O2 consumption, glucose, and no acetic acid production. The kinetic parameters µ(h), YX/S, YP/S, QP, and QX did not differ significantly; however, the kinetic data were superior. Our results suggest that in silico tools allow epitope selection and bioprocess standardization. This system provides cost savings and technological advances for the veterinary vaccine industry.
是家畜和人类中的常见病原体,α-毒素是其发病机制的主要原因。由于它是一种苛求菌,获取α-毒素成本高昂。因此,我们提出了一项计算机模拟研究,以在大肠杆菌BL21 pLysS (DE3)培养物中合成表位。首先,我们使用了搅拌罐生物反应器,在分批培养中干物质产量(DMY)为0.77 g/L,在补料分批培养中为1.03 g/L,且无乙酸产生。在使用无机械搅拌系统进行放大培养时,α-毒素表位1(DE3/Ep1)为0.56 mmol/mL,α-毒素表位2(DE3/Ep2)为0.61 mmol/mL时,DMY更高(1.20 g/L),氧气消耗、葡萄糖情况类似,且无乙酸产生。动力学参数µ(h)、YX/S、YP/S、QP和QX无显著差异;然而,动力学数据更优。我们的结果表明,计算机模拟工具可实现表位选择和生物过程标准化。该系统为兽用疫苗行业节省了成本并带来了技术进步。