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Synthesis and Purification of Mature Human Serum Albumin from E. Coli

Bio/Technology volume 5, pages 1309–1314 (1987)Cite this article

Abstract

Significant synthesis of Human Serum Albumin (HSA) was demonstrated in E. coli using efficient bacterial expression signals. The recombinant HSA is produced essentially as an insoluble aggregate and differs from natural HSA by the presence of a methionine at the N-terminus. However, the Met-HSA could be re-natured, purified and was found to be very similar to the natural protein. A procedure was devised to produce mature HSA starting with the authentic N-terminal sequence from E. coli. By analogy with the maturation of the HSA propeptide, a plasmid expressing a fusion between the first six residues of the bacteriophage λcII protein (Val-Arg-Ala-Asn-Lys-Arg-) and the sequence of mature HSA was constructed. After renaturation, this fusion protein was cleaved by trypsin in vitro to yield a mature refolded HSA with native N-terminal sequence. This protein could not be distinguished by several criteria from authentic natural HSA.

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Author notes

  1. Michael Knapp

    Present address: College of Physicians and Surgeons of Columbia University, Center for Neurobiology and Behavior, 722 West 168th Street, New York, N.Y., 10032, USA

  2. Paolo Sarmientos

    Present address: Dipartimento di Biologia Molecolare Farmitalia-Carlo Erba, Viale Bezzi 24, 20146, Milano, Italia

Authors and Affiliations

  1. Genetica SA, 160, quai de Polangis, 94340, Joinville le Pont, France

    Martine Latta, Michael Knapp, Paolo Sarmientos, Georges Bréfort & Jean-François Mayaux

  2. Rhône-Poulenc Santé, Centre de Recherches de Vitry, 13 quai Jules Guesde, BP14, 94403, Vitry-Sur-Seine CEDEX, France

    Jérôme Becquart, Luc Guerrier & Gérard Jung

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Latta, M., Knapp, M., Sarmientos, P. et al. Synthesis and Purification of Mature Human Serum Albumin from E. Coli. Nat Biotechnol 5, 1309–1314 (1987). https://doi.org/10.1038/nbt1287-1309

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