Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase
Reexamination Certificate
1995-03-07
2003-07-29
Achutamurthy, Ponnathapu (Department: 1652)
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Hydrolase
C435S069100, C435S252300, C435S320100, C435S471000, C510S392000, C536S023200
Reexamination Certificate
active
06599730
ABSTRACT:
TECHNICAL FIELD
The present invention relates to novel enzyme variants useful in a variety of cleaning compositions, and the genes encoding such enzyme variants.
BACKGROUND
Enzymes make up the largest class of naturally occurring proteins. Each class of enzyme generally catalyzes (accelerates a reaction without being consumed) a different kind of chemical reaction. One class of enzymes known as proteases, are known for their ability to hydrolyze (break down a compound into two or more simpler compounds with the uptake of the H and OH parts of a water molecule on either side of the chemical bond cleaved) other proteins. This ability to hydrolyze proteins has been taken advantage of by incorporating naturally occurring and protein engineered proteases as an additive to laundry detergent preparations. Many stains on clothes are proteinaceous and wide-specificity proteases can substantially improve removal of such stains.
Unfortunately, the efficacy level of these proteins in their natural, bacterial environment, frequently does not translate into the relatively unnatural wash environment. Specifically, protease characteristics such as thermal stability, pH stability, oxidative stability and substrate specificity are not necessarily optimized for utilization outside the natural environment of the enzyme.
The amino acid sequence of the protease determines the characteristics of the protease. A change of the amino acid sequence of the protease may alter the properties of the enzyme to varying degrees, or may even inactivate the enzyme, depending upon the location, nature and/or magnitude of the change in the amino acid sequence. Several approaches have been taken to alter the wild-type amino acid sequence of proteases in an attempt to improve their properties, with the goal of increasing the efficacy of the protease in the wash environment. These approaches include altering the amino acid sequence to enhance thermal stability and to improve oxidation stability under quite diverse conditions.
Despite the variety of approaches described in the art, there is a continuing need for new effective variants of proteases useful for cleaning a variety of surfaces.
OBJECTS OF THE PRESENT INVENTION
It is an object of the present invention to provide subtilisin 309 enzyme variants having improved hydrolysis versus the wild-type of the enzyme.
It is also an object of the present invention to provide cleaning compositions comprising these subtilisin 309 enzyme variants.
SUMMARY
The present invention relates to subtilisin 309 variants having a modified amino acid sequence of wild-type subtilisin 309 amino acid sequence, the wild-type amino acid sequence comprising a first loop region, a second loop region, a third loop region, a fourth loop region, a fifth loop region and a sixth loop region; wherein the modified amino acid sequence comprises different amino acids than that occurring in wild-type subtilisin 309 (i.e., substitution) at specifically identified positions in one or more of the loop regions whereby the subtilisin 309 variant has decreased adsorption to, and increased hydrolysis of, an insoluble substrate as compared to the wild-type subtilisin 309. The present invention also relates to DNA sequences encoding such subtilisin 309 variants. The present invention also relates to compositions comprising such subtilisin 309 variants for cleaning a variety of surfaces.
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Barnett Bobby Lee
Brode, III Philip Frederick
Ghosh Chanchal Kamur
Rubingh Donn Nelton
Achutamurthy Ponnathapu
Lewis Len
Miller Steve W.
Moore William W.
Procter & Gamble Company
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