Appeal No. 2005-1745
Application No. 09/161,680
12. A method for generating a new catalytic activity in an enzyme, comprising
the steps of:
a) introducing a DNAsequence 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 transformed 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 XL1-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 comprises at least one enzyme
substrate to recognize the newly generated catalytic activity in the
enzyme, 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 and
phytases.
20. The method of claim 13, wherein the lipase is selected from the group of
lipases consisting of Pseudomonas cepacia lipases PS, Pseudomonas
cepacia lipase AH, acylase, Rhizopus delamar lipase, Rhizopus javanicus
lipase, Candida rugosa lipase, Mucor javanicus lipase, Penicillium roquefortii
lipase, Penicillium cyclopium lipase, Chromobacterium viscosum lipase,
Rhizomucor miehei lipase, Humicola lanuginosa lipase, Candida antarctica
lipase B and Candida antarctica lipase A.
2
Page: Previous 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Next
Last modified: November 3, 2007