Transgenic soybean seed as protein expression system: aqueous extraction of recombinant beta-glucuronidase.
Soybean is one of the plant species with potential to be used as seed-based bioreactor. As part of the downstream processing (DSP) of this technology, extraction is a key step, since it defines the composition of the solution from which the recombinant product will be purified. In the present work, the characteristics of soybean seeds used as a bioreactor were evaluated from a process engineering standpoint through analysis of the influence of pH and ionic strength on the extraction of recombinant beta-glucuronidase (rGUS). Concentrations of recombinant protein and native soybean compounds were analyzed and compared with similar data from extraction studies using transgenic corn seeds as bioreactor. Efficient rGUS extraction was obtained at pH of around 5.5 with no addition of salt. Soybean seed extracts had lower levels of co-extracted native compounds, than corn seed extracts, and should be considered as a potential plant bioreactor in terms of DSP.
LEBp: Laboratório de Engenharia de Bioprocessos, Departamento de Processos Biotecnológicos, Faculdade de Engenharia Química, Universidade Estadual de Campinas, CP 6066, CEP 13083-970, Campinas, SP, Brazil.
Goran
G
Robić
Cristiane S
CS
Farinas
Elíbio L
EL
Rech
Everson A
EA
Miranda
D019149:Bioreactors; D018506:Gene Expression Regulation, Plant; D005966:Glucuronidase / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D010940:Plant Proteins / Q000096:biosynthesis* / Q000235:genetics; D030821:Plants, Genetically Modified / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis*; D012639:Seeds / Q000235:genetics; D013025:Glycine max / Q000235:genetics*; D003313:Zea mays / Q000235:genetics
D010940:Plant Proteins; D011994:Recombinant Proteins; D005966:Glucuronidase