Cultivation of rE. coli in airlift bioreactor: development of strategies for optimization of biomass production

• Grant number: 11/16605-3
• Support type: Scholarships in Brazil - Master
• Effective date (Start): March 01, 2012
• Effective date (End): February 28, 2014
• Field of knowledge: Engineering - Chemical Engineering - Chemical Process Industries
• Principal researcher: Roberto de Campos Giordano
• Grantee: Gilson Campani Junior
• Home Institution: Centro de Ciências Exatas e de Tecnologia (CCET). Universidade Federal de São Carlos (UFSCAR). São Carlos , SP, Brazil
• Associated research grant: 08/05207-4 - Pneumococcal conjugate vaccine: capsular polysaccharide - pneumococcal surface protein A, AP.TEM

Abstract

Due to the large amount of available information concerning biochemical, physiological and genetic characterization as well as to the easy manipulation and cultivation, several proteins with industrial and therapeutic application are produced by genetically modified Escherichia coli. For this reason, the literature is plenty of information about strategies for increasing biomass and recombinant protein production using rE. coli fed batch cultivation with stirred and aerated tank bioreactors, including also strategies for bioprocess control and automation. On the other hand, despite of its broad use for filamentous fungi cultivation, few works reporting rE. coli in airlift type pneumatic reactors are found. These bioreactors show several advantages regarding the conventional ones, such as simple construction, less risk of contamination, ease for scaling up and efficient gas-liquid dispersion with low energy consumption. Within this context, the present MSc proposal aims at developing new operation strategies for airlift that optimize biomass production in rE. coli cultures. The activities to be carried out include, initially, the design and construction of an airlift bioreactor suitable for fed-batch operation. In the next step, the software SuperSys_HCDC, developed for supervision of high density E. coli cultures in stirred tank bioreactors, will be adequate for monitoring and controlling the previously built airlift bioreactor. Finally, batch and fed-batch cultures of E. coli BL21(DE3)pET37b+/PspA4Pro, expressing a fragment of pneumococcal surface protein A (PspA) from clado 4 (PspA4Pro), will be conducted with this airlift bioreactor integrated to the control and monitoring system. Variables such as air flow rate, air enrichment with oxygen, temperature e reactor pressure will be manipulated, one by one or together, aiming at the identification of the operation conditions that will lead to maximum biomass formation. Broth rheology will also be assessed. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

CAMPANI, GILSON; ARRUDA RIBEIRO, MARCELO PERENCIN; LUPERNI HORTA, ANTONIO CARLOS; GIORDANO, ROBERTO CAMPOS; BADINO, ALBERTO COLLI; ZANGIROLAMI, TERESA CRISTINA. Oxygen transfer in a pressurized airlift bioreactor. Bioprocess and Biosystems Engineering, v. 38, n. 8, p. 1559-1567, AUG 2015. (08/05207-4, 11/16605-3)

FIGUEIREDO, DOUGLAS B.; CARVALHO, ENEAS; SANTOS, MAURICIO P.; KRASCHOWETZ, STEFANIE; ZANARDO, RAFAELA T.; CAMPANI, JR., GILSON; SILVA, GABRIEL G.; SARGO, CINTIA R.; HORTA, ANTONIO CARLOS L.; GIORDANO, ROBERTO DE C.; et al. Production and purification of an untagged recombinant pneumococcal surface protein A (PspA4Pro) with high-purity and low endotoxin content. Applied Microbiology and Biotechnology, v. 101, n. 6, p. 2305-2317, MAR 2017. (15/06255-6, 12/04858-7, 11/16605-3)