title: Alternate carbohydrate and nontraditional inducer leads to increased productivity of a collagen binding domain fusion protein via fed-batch fermentation.
abstract: The production of collagen binding domain fusion proteins is of significant importance because of their potential as therapeutic biomaterials. It was previously reported that the expression of collagen-binding domain fusion proteins in Escherichia coli was higher when expressed using lactose as an inducer and chemically defined growth media on a shake flask scale. In an effort to further investigate factors that affect expression levels on a fed-batch scale, alternative induction techniques were tested in conjunction with fed-batch fermentation. In this paper, we discuss ten fed-batch fermentation experiments utilizing either glucose or glycerol feed and using lactose or isopropyl-β-d-thiogalactopyranoside (IPTG) as an induction source. It was found that glycerol-fed fermentations induced with lactose allowed for greater expression of target protein, though lesser cell densities were achieved.
authors.affiliation: Ralph E. Martin Department of Chemical Engineering, 3202 Bell Engineering Center, University of Arkansas, Fayetteville, AR 72701, USA.
authors.forename: McKinzie
authors.initials: M
authors.lastname: Fruchtl
authors.affiliation: Department of Chemistry and Biochemistry, University of Arkansas, 119 Chemistry Building, University of Arkansas, Fayetteville, AR 72701, USA.
authors.forename: Joshua
authors.initials: J
authors.lastname: Sakon
authors.affiliation: Ralph E. Martin Department of Chemical Engineering, 3202 Bell Engineering Center, University of Arkansas, Fayetteville, AR 72701, USA. Electronic address: rbeitle@uark.edu.
authors.forename: Robert
authors.initials: R
authors.lastname: Beitle
mesh_terms: D000085:Acetates / Q000032:analysis; D061249:Batch Cell Culture Techniques / Q000379:methods*; D018533:Biomass; D015153:Blotting, Western; D002241:Carbohydrates / Q000494:pharmacology*; D003094:Collagen / Q000737:chemistry*; D005285:Fermentation / Q000187:drug effects*; D005947:Glucose / Q000032:analysis; D005990:Glycerol / Q000032:analysis; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology*; D007785:Lactose / Q000032:analysis; D010957:Plasmids / Q000378:metabolism; D000072417:Protein Domains; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D013997:Time Factors
chemical_list: D000085:Acetates; D002241:Carbohydrates; D011993:Recombinant Fusion Proteins; D007544:Isopropyl Thiogalactoside; D003094:Collagen; D005947:Glucose; D007785:Lactose; D005990:Glycerol
keywords: Autoinduction; Collagen binding domain fusion proteins; Fed-batch fermentation