Improving bioreactor production of a recombinant glycoprotein in Escherichia coli : Effect of specific growth rate on protein glycosylation and specific productivity

CAILLAVA, ANA J.; ORTIZ, GASTÓN E.; MELLI, LUCIANO J.; UGALDE, JUAN E.; CIOCCHINI, ANDRÉS E.; COMERCI, DIEGO J.

BIOENGINEERING AND BIOTECHNOLOGY

JOHN WILEY & SONS INC

Año: 2019 vol. 116 p. 1427 - 1438

0006-3592

In the last decades bacterial glycoengineering emerged as a new field as the result ofthe ability to transfer the Campylobacter jejuni N‐ glycosylation machinery intoEscherichia coli for the production of recombinant glycoproteins that can be used asantigens for diagnosis, vaccines, and therapeutics. However, the identification ofcritical parameters implicated in the production process and its optimization to jumpto a productive scale is still required. In this study, we developed a dual expressionglycosylation vector for the production of the recombinant glycoprotein AcrA‐O157,a novel antigen that allows the serodiagnosis of the infection with enterohemorrhagicE. coli O157 in humans. Volumetric productivity was studied in different culturemedia and found that 2xYP had 6.9‐fold higher productivity than the extensively usedLB. Subsequently, bioreactor batch and exponential‐fed‐batch cultures were designedto determine the influence of the specific growth rate (μ) on AcrA‐O157 glycosylationefficiency, production kinetics, and specific productivity. At μmax, AcrA glycosylationwith O157‐polysaccharide and the specific synthesis rate were maximal, constitutingthe optimal physiological condition for AcrA‐O157 production. Our findings should beconsidered for the design, optimization, and scaling up of AcrA‐O157 production andother recombinant glycoproteins attractive for industrial applications.