title: Effect of environmental conditions on the expression levels of a recombinant 11S amaranth globulin in Escherichia coli.
abstract: The expression in Escherichia coli strain Rosetta of the recombinant acidic subunit from the 11S amaranth seed storage protein (protein ACM3) was studied at flask and at bioreactor levels. This subunit was modified by inserting four Val-Tyr antihypertensive peptides in tandem into its third variable region and also with the tripeptide Ile-Pro-Pro in the Cterminal region. Flasks experiments allowed us to define the best conditions for the preparation and expression and accumulation of the protein ACM3, including the certainty of its presence within the cells especially as an insoluble fraction. The effects of cultivation temperature, aeration rate and agitation speed on the production of the protein ACM3 was tested in a 5-L batch bioreactor. Applying response surface methodology (RSM) we found that the aeration rate was the most significant factor affecting in a positively way the production yields and productivity of the recombinant protein. Temperature had effect only in conjunction to aeration. The highest recombinant acidic subunit concentration (747 mg L-1) and the highest productivity (186 mg L-1 h-1) were attained in 4 h of cultivation when the factors evaluated were controlled at its central values: 0.1 vvm, 300 rpm, and 30.5° C. Results from this study indicate that RSM is an effective technique to maximize the production of this recombinant protein.
authors.affiliation: Cinvestav-IPN, Irapuato Unit, Km 9.6 Libramiento Norte Carr. Irapuato-Leon, 36821, Irapuato, Gto., Mexico.
authors.forename: Hypatia
authors.initials: H
authors.lastname: Arano-Varela
authors.forename: Jorge
authors.initials: J
authors.lastname: Domínguez-Domínguez
authors.forename: Octavio
authors.initials: O
authors.lastname: Paredes-López
mesh_terms: D000704:Analysis of Variance; D019149:Bioreactors / Q000382:microbiology*; D004926:Escherichia coli / Q000378:metabolism*; D005285:Fermentation; D005916:Globulins / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000096:biosynthesis; D012044:Regression Analysis; D015203:Reproducibility of Results; D055314:Seed Storage Proteins / Q000096:biosynthesis*
chemical_list: D005916:Globulins; D011994:Recombinant Proteins; D055314:Seed Storage Proteins