Appeal No. 2005-1383
Application No. 09/364,847
pp. 226-27. As pointed out by the examiner, Bülow still further discloses different
methods of “creating proximity between the enzymes” (p. 227, col. 1, first complete
para.) which include gene fusions which “maintain the monomeric character of the
native enzymes” (id., col. 2, first complete para.) by “joining the structural genes of two
enzymes: the translational stop signal at the 3’ end of the first gene is removed and
ligated in-frame to the ATG start codon of the second gene” (id., col. 1, para. 3). Bülow
still further discloses that through experience, it has “found that fairly short linkers (two
to ten amino acid residues) [between the fused enzymes] are optimal.” Id., p. 230,
col. 1, first complete para.; see also, Figure 1c and Figure 3. Bülow exemplifies several
fusion proteins consisting of catalytically active enzymes which act on substrate in
successive reactions. See, the β-galactosidase-galactokinase fusion protein
(pp. 227-228), the β-galactosidase-galactose dehydrogenase fusion protein (p. 228,
cols. 1-2), and the bovine P-450s and yeast reductase fusion protein (p. 229, col. 2).
Not only does Bülow exemplify several fusion proteins, but the publication reports that
“[o]ver the past few years a variety of artificial bifunctional enzymes have been prepared
by gene fusion in vitro.” Id., p. 227, col. 2, last para.
Accordingly, in view of the teachings of Peoples with respect to the construction
of (i) plasmids comprising two catalytically active enzymes in the PHA biosynthetic
pathway which act on a substrate in successive reactions (see, e.g., col. 22,
lines 26-30; Figure 3); and (ii) protein fusions comprising catalytically active enzymes
involved in the PHA biosynthetic pathway (col. 23, lines 14-24), and the teachings of
Bülow as to the construction of bifunctional enzymes comprising two enzymes involved
in successive reactions, the advantages of said constructions, and that said
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