Protein-based materials biosynthesis

The use of recombinant DNA technology, that is, biosynthesis or bioproduction, of protein-based materials may be considered in three main stages (1) gene construction, (2) cell transformation, and (3) fermentation of micro-... 

Ferrari, F.A. and Cappello, J. (1997) Biosynthesis of Protein Polymers in Protein-Based Materials, Biorkauser, Boston. 

Protegrin derivatives, 18 260 Protegrins, 18 260-261 properties of, 18 261 Protein. See also Proteins extraction of, 26 474 in cereal grains, 26 275-276 Proteinaceous materials, as membrane foulants, 21 664 Protein adsorption, 12 136-137 Protein affinity libraries, 12 516-517 Protein-based chiral phases, 6 89-90 Protein-based microarrays, 16 382 Protein biosynthesis, 20 450... 

Non-natural amino acids can be incorporated into peptides and polypeptides via several different methodologies. Solid-phase peptide synthesis (SPPS) is a straightforward method for incorporation of non-natural amino acids and allows the incorporation of essentially any amino acid but is limited by the size of the peptides produced 18). Suppression-based strategies, both in vitro and in vivo, have been developed for site specific incorporation of diverse set non-natural amino acids into natural and synthetic polypeptides 19). Alternatively, auxotrophic expression hosts have been used for multisite incorporation of nonnatural amino acid in protein polymers, where multiple natural amino acids of one type can be replaced with non-natural analogues during protein biosynthesis (20, 21). Multisite incorporation of non-natural amino acids in the synthesis of protein polymeric materials facilitates chemical modification at multiple sites and can modulate the physical properties of the protein polymers (22). 

A few examples are discussed in Chapter 9 wherein the future will see the biosynthesis of biodegradable protein-based thermoplastics, materials to prevent postsurgical adhesions, temporary functional scaffoldings to direct tissue reconstruction, and drug delivery devices for new drug release regimens. 

As introduced in Chapter 1, the present chapter constitutes Assertion 4 The Applications Assertion of the book. Production and purification are first addressed, as they obviously make up the initial enabling steps in moving toward applications of any materials. The most surefooted path toward materials applications of protein-based polymers, however, intertwines issues of production and purification through a combination of the two methods of preparation—chemical synthesis and biosynthesis. Chemical synthesis proved the biocompatibility of elastic protein-based polymers and therefore opened the door to medical applications. Demonstration of the biocompatibility of the chemically synthesized product made clear the purification required of elastic protein-based polymers produced by E. coli if unlimited medical applications were to be possible. Chemical synthesis also provided a faster route to diverse polymer compositions, which allowed... 

In one of the early experiments designed to elucidate the genetic code, Marshall Nirenberg of the U.S. National Institutes of Health (Nobel Prize in physiology or medicine, 1968) prepared a synthetic mRNA in which all the bases were uracil. He added this poly(U) to a cell-free system containing all the necessary materials for protein biosynthesis. A polymer of a single amino acid was obtained. What amino acid was polymerized ... 

Protein polymers based on Lys-25 were prepared by recombinant DNA (rDNA) technology and bacterial protein expression. The main advantage of this approach is the ability to directly produce high molecular weight polypeptides of exact amino acid sequence with high fidelity as required for this investigation. In contrast to conventional polymer synthesis, protein biosynthesis proceeds with near-absolute control of macromolecular architecture, i.e., size, composition, sequence, topology, and stereochemistry. Biosynthetic polyfa-amino acids) can be considered as model uniform polymers and may possess unique structures and, hence, materials properties, as a consequence of their sequence specificity [11]. Protein biosynthesis affords an opportunity to completely specify the primary structure of the polypeptide repeat and analyze the effect of sequence and structural uniformity on the properties of the protein network. 

Since a-amino-acids serve as starting materials for the synthesis of protein and the elaboration of many plant alkaloids, there must be a sharing of any amino-acid which is required for both of these activities. The extent to which this happens has been the subject of a new study in one particular plant, Lophophora williamsii, which produces isoquinoline and j8-phenethylamine alkaloids. These bases are derived from the a-amino-acid tyrosine and the results from feeding L-[f/- C]tyrosine indicate that this amino-acid is incorporated into the alkaloids approximately three times more efficiently than into protein. Only the L-isomer was examined and one wonders what the results with D-tyrosine would be in the light of the known preference for particular optical isomers of lysine in pipecolic acid and piperidine alkaloid biosynthesis. 

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