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Chemical and biosynthetic approaches to the production of novel polypeptide materials

McGrath, Kevin P. and Tirrell, David A. and Kawai, Michio and Mason, Thomas L. and Fournier, Maurille J. (1990) Chemical and biosynthetic approaches to the production of novel polypeptide materials. Biotechnology Progress, 6 (3). pp. 188-192. ISSN 8756-7938. doi:10.1021/bp00003a004.

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Three approaches to the synthesis of the repetitive copolypeptide [-(GlyAla)_3- GlyProGlu-]_n (1) are described. Direct chemical synthesis of 1 via classical solution methods required 18 steps and afforded a polydisperse product with an average molecular weight of less than 10 000. Two alternative genetic strategies were also explored. In the first, chemically synthesized DNA oligomers were self-ligated to produce a population of multimers, which were fitted with translational start and stop signals and inserted into an expression plasmid containing the λ P_L promoter and a synthetic ribosome binding site. Transformation of E. coli led to the isolation of a stable recombinant plasmid carrying an insert encoding 12 repeats of sequence 1. Attempts to identify polypeptide 1 after induction of transformed cultures were unsuccessful. A second strategy, generating a tripartite derivative of sequence 1 carrying short N- and C-terminal extensions, afforded excellent yields of product. The relative merits of chemical and genetic approaches to repetitive polypeptide materials are discussed.

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Tirrell, David A.0000-0003-3175-4596
Additional Information:© 1990 American Chemical Society and American Institute of Chemical Engineers. Accepted January 17, 1990. We acknowledge the contributions of Drs. Eiji Yashima and Yasumasa Fukushima, and Howard Creel, for their work on the chemical synthesis of the repetitive polymer. We are also indebted to Richard Lempicki for technical assistance and advice throughout the course of this work. Dr. Joseph Cappello of Protein Polymer Technologies, Inc., provided the vector p937.51, as well as valuable advice regarding the successful gene construction described briefly herein. We are grateful to the Center for UMass-Industry Research on Polymers (CUMIRP) for financial assistance and to Mitsubishi Chemical Industries for granting a leave of absence for M.K.
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Center for UMass-Industry Research on Polymers (CUMIRP)UNSPECIFIED
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Official Citation:McGrath, K. P., Tirrell, D. A., Kawai, M., Mason, T. L. and Fournier, M. J. (1990), Chemical and Biosynthetic Approaches to the Production of Novel Polypeptide Materials. Biotechnol Progress, 6: 188–192. doi: 10.1021/bp00003a004
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ID Code:53526
Deposited By: Anne Hormann
Deposited On:22 Jan 2015 21:53
Last Modified:10 Nov 2021 20:03

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