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) Bacterial cultures such as Pseudomonas oleovorans and Ralstonia eutropha have been used to produce PHA polymers in the laboratory The polyester structure can be varied to some extent by controlling the organic feed Biopol is a small-scale commercial product a copolymer of 3-hydroxybutanoate and 3-hydroxypentanoate repeat units Such polymers are of interest because they are environmentally degradable and offer an alternative to recycling the large volumes of nondegradable polymer wastes The mechanism for bacterial synthesis of PHA is not the simple dehydration reaction between alcohol and carboxyl groups It is more complicated and involves the coenzyme A thioester derivative of the hydroxyalkanoic acid monomer (produced from the organic feedstock available to the bacteria) [Kamachi et al, 2001] Growth involves an acyl transfer reaction catalyzed by the enzyme PHA synthase (also called a polymerase) [Blei and Odian, 2000].



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The reaction is not a step polymerization but is a chain polymerization with the characteristics of a living polymerization [Su et al, 2000] (Secs 3-15, 5-2g, Chap 7) 2-17a-2 In Vitro (outside Living Cells).





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The in vivo production of commodity polymers has been limited by the dif culties of controlling these processes, including the control of the reaction conditions required to maintain a bacterial culture and the conditions needed to harvest the polymer product. In addition, the in vivo method, although applicable for a few polymers, is not useful for the full range of polymers produced by the typical chemical processes. An alternate to the in vivo method is the use of enzymes to catalyze in vitro polymerizations. In vitro enzymatic polymerizations are enzyme-catalyzed polymerizations taking place outside cells, that is, taking place in glass and metal reactors. Biochemists carry out in vitro enzymatic polymerizations using the same reactants present in living cells for the purpose of understanding the reactions within living cells [Kamachi et al., 2001]. For polymer chemists, the major effort is the use of enzymes extracted from living cells to carry out in vitro polymerizations of nonnatural substrates to produce polymers for industrial use [Gross et al., 2001; Kobayashi, 1999; Kobayashi et al., 2001]. Lipases are enzymes that catalyze the in vivo hydrolysis of lipids such as triacylglycerols. Lipases are not used in biological systems for ester synthesis, presumably because the large amounts of water present preclude ester formation due to the law of mass action which favors hydrolysis. A different pathway (using the coenzyme A thioester of a carboxylic acid and the enzyme synthase [Blei and Odian, 2000]) is present in biological systems for ester formation. However, lipases do catalyze the in vitro esteri cation reaction and have been used to synthesize polyesters. The reaction between alcohols and carboxylic acids occurs in organic solvents where the absence of water favors esteri cation. However, water is a by-product and must be removed ef ciently to maximize conversions and molecular weights.

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where rand s stand for 1, 2, or 3. Dirac's solution was to pick them as matrices and the wavefunction as a spinor. As is well known [159], the lowest possible dimension for these spinors is four. Moreover, Dirac's choice of spinors and matrices only allows even dimensions. This is all right for electrons 14 but is not what we need for light. The selection rules observed for absorption and emission of light by matter tell us that the light quantum must have an intrinsic angular momentum (Le., spin) of one unit of Planck's constant [472]. Lanczos [383, 384] developed a generalization of Dirac's approach to factorize (3.19). Lanczos's idea was to use biquatemions instead of matrices and spinors}5 Biquatemions are complex quatemions, and quatemions are, in a nutshell, a fourdimensional generalization of complex numbers introduced by Hamilton in 1844 [263, 264]. Quatemions can be thought as a scalar plus an axial vector [567]. The

The reaction mechanism for the polymerization of a hydroxyalkanoic acid (Eqs. 2-243 through 2-246) is a chain polymerization, often called an activated monomer polymerization. The active site of lipase is its serine a-amino acid unit, which contains a hydroxyl group. The acyl carbon of the hydroxyalkanoic acid undergoes nucleophilic attack by the hydroxyl group of serine to form lipase-activated monomer (Eq. 2-243). Initiation consists of reaction

4 1 7

13There is no escape from the double sign in the end. 14They turn out to have spin 112. which can be described by a four component spinor and those 4 x 4 matrices. The extra two dimensions are needed to account for the positrons. which appear in the theory because of the negative energy solutions that can be traced back to the minus sign in (3.25) coming back to haunt us. 15For a very readable introduction to this way of writing field equations. see [427].

2-244 2-243

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