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首页> 外文期刊>Brazilian Journal of Microbiology >Production of outer membrane vesicles (OMV) in batch cultivation of Neisseria meningitidis serogroup B
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Production of outer membrane vesicles (OMV) in batch cultivation of Neisseria meningitidis serogroup B

机译：脑膜炎奈瑟氏球菌血清群B的分批培养中的外膜囊泡（OMV）的生产

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Meningococcal disease is an important cause of death and morbidity throughout the world. Nearly 330,000 cases and 35,000 deaths occur yearly. Neisseria meningitidis, serogroup B strain N.44/89, is prevalent in Brazil. Its outer membrane vesicles (OMV) with iron regulated proteins (IRP) are released to the culture medium and are used as antigen for vaccine production. In order to have knowledge about the kinetic parameters, especially the final OMV concentration values, 20-h batch cultivations were carried out in Catlin medium with iron restriction. Process conditions comprised: 7 L bioreactor, 36oC, 0.5 atm, overlay air flowrate of 1 L/min, agitation varying from 250 rpm to 850 rpm and dissolved oxygen control set at 10% of saturation condition. Biomass was determined by optical density at 540 nm and dry weight. Glycerol, lactate, pH and dissolved oxygen were measured from samples taken during cultivation. Outer membrane vesicle (OMV) concentration was determined by Lowry's method after ultracentrifugation. IRP presence was verified by SDS-PAGE. Highest biomass value, corresponding to the highest initial lactate concentration (7.84 g/L) was achieved at the 9th hour process time corresponding to 1.0 g/L dry biomass and 2.3 optical density at 540 nm. Lactate consumption was directly related to cell growth (yield factor: 0.24 g dry biomass / g lactate). Glycerol concentration in the medium did not change significantly during the process. OMV concentration reached the highest value of 80 mg/L at end cultivation time. The obtained results suggest that lactate is a main limiting growth factor and the maximum amount of antigen is obtained during stationary growth and cell death phases.
机译：脑膜炎球菌病是世界范围内死亡和发病的重要原因。每年有近330,000例病例和35,000例死亡。脑膜炎奈瑟氏球菌，血清群B株N.44 / 89，在巴西很流行。其具有铁调节蛋白（IRP）的外膜囊泡（OMV）被释放到培养基中，并用作生产疫苗的抗原。为了了解动力学参数，尤其是最终OMV浓度值，在含铁限制的Catlin培养基中进行了20小时的分批培养。工艺条件包括：7 L生物反应器，36oC，0.5 atm，1 L / min的重叠空气流量，搅拌速度从250 rpm到850 rpm不等，溶解氧控制设置为饱和条件的10％。通过540nm处的光密度和干重确定生物量。从培养期间采集的样品中测量甘油，乳酸，pH和溶解氧。超速离心后，通过Lowry法测定外膜囊泡（OMV）浓度。通过SDS-PAGE验证了IRP的存在。在第9小时的处理时间获得了最高的生物量值，对应于最高的初始乳酸浓度（7.84 g / L），对应于1.0 g / L的干燥生物量和540 nm处的2.3光学密度。乳酸消耗与细胞生长直接相关（产量因子：0.24 g干生物质/ g乳酸）。在此过程中，培养基中甘油的浓度没有明显变化。在培养结束时，OMV浓度达到最高值80 mg / L。获得的结果表明，乳酸是主要的限制生长因子，并且在稳定的生长和细胞死亡阶段获得了最大量的抗原。

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