title: Expression and purification of short hydrophobic elastin-like polypeptides with maltose-binding protein as a solubility tag.
abstract: Elastin-like polypeptides (ELPs) are biodegradable polymers with interesting physico-chemical properties for biomedical and biotechnological applications. The recombinant expression of hydrophobic elastin-like polypeptides is often difficult because they possess low transition temperatures, and therefore form aggregates at sub-ambient temperatures. To circumvent this difficulty, we expressed in Escherichia coli three hydrophobic ELPs (VPGIG)n with variable lengths (n=20, 40, and 60) in fusion with the maltose-binding protein (MBP). Fusion proteins were soluble and yields of purified MBP-ELP ranged between 66 and 127mg/L culture. After digestion of the fusion proteins by enterokinase, the ELP moiety was purified by using inverse transition cycling. The purified fraction containing ELP40 was slightly contaminated by traces of undigested fusion protein. Purification of ELP60 was impaired because of co-purification of the MBP tag during inverse transition cycling. ELP20 was successfully purified to homogeneity, as assessed by gel electrophoresis and mass spectrometry analyses. The transition temperature of ELP20 was measured at 15.4°C in low salt buffer. In conclusion, this method can be used to produce hydrophobic ELP of low molecular mass.
authors.affiliation: Univ. Bordeaux, LCPO, UMR 5629, F-33600 Pessac, France; CNRS, LCPO, UMR 5629, F-33600 Pessac, France; Bordeaux INP, LCPO, UMR 5629, F-33600 Pessac, France; Institut Européen de Chimie et Biologie, F-33600 Pessac, France.
authors.forename: Laure
authors.initials: L
authors.lastname: Bataille
authors.affiliation: Univ. Bordeaux, BPRVS, EA4135, F-33000 Bordeaux, France; Bordeaux INP, BPRVS, EA4135, F-33000 Bordeaux, France.
authors.forename: Wilfrid
authors.initials: W
authors.lastname: Dieryck
authors.affiliation: Univ. Bordeaux, BPRVS, EA4135, F-33000 Bordeaux, France; Bordeaux INP, BPRVS, EA4135, F-33000 Bordeaux, France.
authors.forename: Agnès
authors.initials: A
authors.lastname: Hocquellet
authors.affiliation: Univ. Bordeaux, BPRVS, EA4135, F-33000 Bordeaux, France; Bordeaux INP, BPRVS, EA4135, F-33000 Bordeaux, France.
authors.forename: Charlotte
authors.initials: C
authors.lastname: Cabanne
authors.affiliation: Univ. Bordeaux, CBMN, UMR 5248, F-33600 Pessac, France; CNRS, CBMN, UMR5248, F-33600 Pessac, France; Bordeaux INP, CBMN, UMR5248, F-33600 Pessac, France.
authors.forename: Katell
authors.initials: K
authors.lastname: Bathany
authors.affiliation: Univ. Bordeaux, LCPO, UMR 5629, F-33600 Pessac, France; CNRS, LCPO, UMR 5629, F-33600 Pessac, France; Bordeaux INP, LCPO, UMR 5629, F-33600 Pessac, France.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Lecommandoux
authors.affiliation: Univ. Bordeaux, BPRVS, EA4135, F-33000 Bordeaux, France; Bordeaux INP, BPRVS, EA4135, F-33000 Bordeaux, France. Electronic address: bertrand.garbay@ipb.fr.
authors.forename: Bertrand
authors.initials: B
authors.lastname: Garbay
authors.affiliation: Univ. Bordeaux, LCPO, UMR 5629, F-33600 Pessac, France; CNRS, LCPO, UMR 5629, F-33600 Pessac, France; Bordeaux INP, LCPO, UMR 5629, F-33600 Pessac, France; Institut Européen de Chimie et Biologie, F-33600 Pessac, France.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Garanger
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D040761:Biomimetic Materials; D004549:Elastin / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D004765:Enteropeptidase / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D057927:Hydrophobic and Hydrophilic Interactions; D058619:Maltose-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D010957:Plasmids / Q000737:chemistry* / Q000378:metabolism; D059748:Proteolysis; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D012995:Solubility; D044366:Transition Temperature
chemical_list: D029968:Escherichia coli Proteins; D058619:Maltose-Binding Proteins; D010455:Peptides; D011993:Recombinant Fusion Proteins; D004549:Elastin; D004765:Enteropeptidase
keywords: Elastin-like polypeptides; Maltose-binding protein transition temperature; Mass spectrometry; Precision polymers; Recombinant expression