Appeal No. 2005-1745
Application No. 09/161,680
24. A method for generating a new catalytic activity in an enzyme, wherein the
new catalytic activity is within the same International Union of Biochemistry
class as the enzyme’s original catalytic activity, comprising the steps of:
a) introducing a DNA sequence coding for the enzyme into the
Escherichia coli strain XL1-Red, or into a functional derivative thereof
which is also an E.coli strain carrying the genetic markers relA1,
mutS, mutT, and mutD5, and having an increased mutation rate,
b) incubating the transformer Escherichia coli strain XL1-Red or its
functional derivative to generate mutations in the DNA sequence,
c) transferring the mutated DNA sequence from the transformed
Escherichia coli strain SX1-Red or its functional derivative to a
microorganism which has no enzyme activity which would impede
selection,
d) incubating this microorganism to detect the new catalytic activity in at
least one selection medium which ocmprises at least one enzyme
substrate to recognize the newly generated catalytic activity, with or
without other indicator substances, and
e) selecting the microorganisms which show the newly generated catalytic
activity, said microorganisms in steps c), d) and e) being a member
selected from the group consisting of bacteria, fungi and yeasts, wherein
the enzyme is selected from the group consisting of lipases, amidases,
nitrilases, ether hydrolases, peroxidases, glycosidases, phytases, and
esterases selected from the group consisting of Pseudomonas
fluorescens esterase, pig liver esterase and Thermoanaerobium brockii
esterase.
The examiner does not rely on any references in rejecting the claims under the
first and second paragraphs of 35 U.S.C. § 112.
The claims stand rejected as follows:
I. Claims 20 and 26 stand rejected under 35 U.S.C. § 112, second paragraph, as being
indefinite for failing to particularly point out and distinctly claim the subject matter
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