Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Isomerase
Reexamination Certificate
2000-01-24
2002-04-16
Prouty, Rebecca E. (Department: 1652)
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Isomerase
C435S069100, C435S183000, C435S200000, C435S252300, C435S320100, C536S023200, C800S295000
Reexamination Certificate
active
06372477
ABSTRACT:
The present invention relates to an enzyme. In addition, the present invention relates to a nucleotide sequence coding for the enzyme. The present invention also relates to one or more uses of the enzyme.
It is known that it is desirable to direct expression of a nucleotide sequence of interest (“NOI”) in certain tissues of an organism, such as a filamentous fungus (e.g.
Aspergillus niger
) or even a plant crop. The resultant protein or enzyme may then be used in industry. Alternatively, the resultant protein or enzyme may be useful for the orgasm itself. For example it may be desirable to produce crop protein products with an optimised amino acid composition and so increase the nutritive value of a crop. For example, the crop may be made more useful as a feed. In the alternative, it may be desirable to isolate the resultant protein or enzyme and then use the protein or enzyme to prepare, for example, food compositions. In this regard, the resultant protein or enzyme can be a component of the food composition or it can be used to prepare food compositions including altering the characteristics or appearance of food compositions. It may even be desirable to use the organism such as a filamentous fungus or a crop plant, to express non-plant nucleotide sequences, such as for the same purposes.
The present invention seeks to provide an enzyme that is useful for industry and also a nucleotide sequence coding for same.
According to a first aspect of the present invention there is provided a UDP-galactose epimerase enzyme obtainable from guar.
According to a second aspect of the present invention there is provided a UDP-galactose epimerase enzyme obtainable from guar, wherein the enzyme comprises at least the sequence shown as SEQ ID No. 3 or SEQ ID No. 4, or a variant homologue or fragment thereof.
According to a third aspect of the present invention there is provided a UDP-galactose epimerase enzyme obtainable from guar, wherein the enzyme is encoded by a nucleotide sequence that comprises at least the sequence shown as SEQ ID No. 1 or SEQ ID No. 2, or a variant, homologue or fragment thereof.
According to a fourth aspect of the present invention there is provided a nucleotide sequence that comprises at least the sequence shown as SEQ ID No. 1 or SEQ ID No. 2, or a variant, homologue or fragment thereof, or a sequence complementary thereto.
According to a fifth aspect of the present invention there is provided a UDP-galactose epimerase enzyme which is immunologically reactive with an antibody raised against a purified UDP-galactose epimerase enzyme according to the present invention.
According to a sixth aspect of the present invention there is provided a process of preparing an enzyme according to the present invention comprising expressing a nucleotide sequence according to the present invention.
According to a seventh aspect of the present invention there is provided the use of an enzyme according to the present invention or an enzyme prepared by a process according to the present invention to prepare a foodstuff (such as a feed).
According to an eighth aspect of the present invention there is provided a nucleotide sequence according to the present invention operatively linked to a promoter.
According to a ninth aspect of the present invention there is provided a foodstuff comprising or prepared from the enzyme according to the present invention or an enzyme prepared by a process according to the present invention.
Other aspects of the present invention include: a construct comprising or capable of expressing the present invention; a vector comprising or capable of expressing the present invention; a plasmid comprising or capable of expressing the present invention: a tissue comprising or capable of expressing the present invention; an organ comprising or capable of expressing the present invention; a transgenic organism comprising or capable of expressing the present invention.
Preferably the enzyme comprises the sequence shown as SEQ. I.D. No. 3 or SEQ. ID No. 4, or a variant, homologue or fragment thereof, and the nucleotide sequence encoding same comprises the sequence shown as SEQ. I.D. No. 1 or SEQ ID No. 2 or a variant, homologue or fragment thereof.
Other aspects of the present invention include methods of expressing or allowing the expression of or transforming any one of the nucleotide sequence, the construct, the plasmid, the vector, the cell, the tissue, the organ or the organism, as well as the products thereof.
Further aspects of the present invention include uses of the enzyme for preparing or treating foodstuffs, including animal feed.
Some of the key advantages of the present invention are that it provides an enzyme having UDP-galactose epimerase activity. UDP-galactose epimerase is an important enzyme as inter alia it catalyses the conversion of UDP-D-glucose to UDP-D-galactose. In addition, the enzyme of the present invention may be prepared in certain or specific cells or tissues, such as in just a specific cell or tissue, of an organism, such as a plant or, by way of possible further example, a microorganism. The present invention also provides a nucleotide sequence coding for the enzyme UDP-galactose epimerase that may be expressed preferably in specific cells or tissues, such an those of an organism, such as a plant or, by way of possible further example, a micro-organism.
Also, the present invention provides constructs, vectors, plasmids, cells, tissues, organs and organisms comprising the nucleotide sequence according to the present invention and methods of expressing the same, preferably in specific cells or tissues, such as expression in just a specific cell or tissue, of an organism such as a plant or, by way of possible further example, a microorganism.
Preferably, the enzyme of the present invention is used in the preparation of a foodstuff. Typical foodstuffs may include dairy products, meat products, poultry products, fish products and bakery products.
The terms “variant”, “homologue” or “fragment” in relation to the amino acid sequence for the preferred UDP-galactose epimerase enzyme of the present invention include any substitution of, variation of, modification of, replacement of, deletion of or addition of one (or more) amino acid from or to the sequence providing the resultant enzyme has UDP-galactose epimerase activity, preferably having at least the same activity of the enzyme comprising the sequence shown as SEQ ID No. 3 or SEQ ID No. 4. In particular, the term “homologue” covers homology with respect to structure and/or function. With respect to sequence homology, preferably there is at least 75%, more preferably at least 85%, more preferably at least 90% homology to an enzyme comprising the sequence shown as SEQ ID No. 3 or SEQ ID No. 4. More preferably there is at least 95%, more preferably at least 98%, homology to an enzyme comprising the sequence shown as SEQ ID No. 3 or SEQ ID No. 4.
The terms “variant”, “homologue” or “fragment” in relation to the nucleotide sequence coding for the UDP-galactose epimerase enzyme of the present invention include any substitution of, variation of, modification of, replacement of, deletion of or addition of one (or more), nucleic acid from or to the sequence providing the resultant nucleotide sequence codes for or is capable of coding for an enzyme having UDP-galactose epimerase activity, preferably having at least the same activity of the enzyme comprising the sequence shown as SEQ ID No. 1 or SEQ ID No. 2. In particular, the term “homologue” covers homology with respect to structure and/or function providing the resultant nucleotide sequence codes for or is capable of coding for an enzyme having UDP-galactose epimerase activity. With respect to sequence homology, preferably there is at least 75%, more preferably at least 85%, more preferably at least 90% homology to a sequence comprising the sequence shown as SEQ ID No. 1 or SEQ ID No. 2. More preferably there is at, least 95%, more preferably at least 98%, homology to a sequence that comprises the sequence shown as SEQ ID No. 1 or SEQ ID No. 2.
T
Brunstedt Janne
J&thgr;rsboe Morten
Petersen Steen Guldager
Danisco A/S
Prouty Rebecca E.
Rao Manjunath
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