Cloning and expression of a functional core streptavidin in Pichia pastoris: strategies to increase yield.

Autor: Casteluber MC; Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil., Damasceno LM, da Silveira WB, Diniz RH, Passos FJ, Passos FM
Jazyk: angličtina
Zdroj: Biotechnology progress [Biotechnol Prog] 2012 Nov-Dec; Vol. 28 (6), pp. 1419-25. Date of Electronic Publication: 2012 Nov 01.
DOI: 10.1002/btpr.1621
Abstrakt: Streptavidin is widely used as an analytical tool and affinity tag together with biotinylated surfaces or molecules. We report for the first time a simple strategy that yields high biomass of a Pichia pastoris strain containing a methanol induced core streptavidin (cStp) gene. Three factors were evaluated for biomass production: glycerol concentration, aeration, and feed flow rates in a bioreactor. Recycling of recombinant cells, either free or immobilized, was investigated during induction. Concentration of 2.0 M glycerol, feeding flow rate of 0.11 mL min(-1) , and aeration by air injection dispersed with a porous stone combined with agitation at 500 rpm were the set of conditions resulting into maximum biomass yield (150 g L(-1) ). These parameters yielded 4.0 g L(-1) of cStp, after 96 h of induction. Recombinant biomass was recycled twice before being discarded, which can reduce production costs and simplify the process. Immobilized P. pastoris biomass produced 2.94 and 1.70 g L(-1) of cStp in the first and second induction cycle, respectively. Immobilization and recycling of recombinant P. pastoris biomass opens new possibilities as a potential strategy to improve volumetric productivity for heterologous protein expression.
(Copyright © 2012 American Institute of Chemical Engineers (AIChE).)
Databáze: MEDLINE
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