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US 7,166,450 B2Granted
Hyaluronan synthase gene and uses thereof
AssigneeThe Board of Regents of the University of Oklahoma
InventorsPaul H. Weigel, Kshama Kumari, Paul DeAngelis
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Abstract
The present invention relates to a method of producing hyaluronic acid that includes introducing a purified nucleic acid segment having a coding region encoding enzymatically active hyaluronan synthase into a Bacillus strain, and culturing the Bacillus strain under conditions appropriate for the production of hyaluronic acid.
Claims38
A method of producing hyaluronic acid, comprising the steps of:
- introducing a purified nucleic acid segment having a coding region encoding enzymatically active hyaluronan synthase into a Bacillus strain, wherein the coding region encodes an enzymatically active hyaluronan synthase that includes at least two amino acid motifs selected from the group consisting of:
- (i) GKR(E/H)(V/C/A)(M/L/Q)(Y/A)(T/W)(A/G)F(X)3-7 (A/S/D)(W/V/A)(D/N)(V/A/Y)(F/V/I)(L/Q/V)(T/V/L)(V/C/I) DSDTX(L/I)(D/E/Y)(P/E/K)X(A/S/C)XXE (SEQ ID NO:11);
- (ii) (L/M)(S/T/V)(D/S/A)XRY(W/Y/D)XAF(G/N/C) (V/I)ER(A/S)(C/A)QSX(F/T)X(N/C/T)(V/I)(L/Q/S)(C/V) (I/V/C)(S/G)GPL(G/S)XY(K/R) (SEQ ID NO:12);
- (iii) (D/E)X(W/Y)(X)2QXF(L/M)G(X)2(C/V)(S/T) XGDDR(H/R/C)LTN (SEQ ID NO:13);
- and
- (iv) (E/D)(T/S/V)P(X)5(W/Y/F)(L/I)XQQ(T/N) RW(S/T/N)KS(Y/W/F)(F/C)RE (SEQ ID NO:14),
- with X being selected from the group consisting of any amino acid;
- and
- wherein the at least two amino acid motifs are found in the relative order of (i), (ii), (iii), and (iv) from an amino terminus to a carboxyl terminus of the enzymatically active hyaluronan synthase;
- and
- culturing the Bacillus strain under conditions appropriate for the production of hyaluronic acid.
8 more elements
A method of producing hyaluronic acid, comprising the steps of:
- introducing a purified nucleic acid segment having a coding region encoding enzymatically active hyaluronan synthase into a Bacillus strain, wherein the coding region encodes an enzymatically active hyaluronan synthase that includes the amino acid motifs:
- (i) GKR(E/H)(V/C/A)(M/L/Q)(Y/A)(T/W)(A/G)F(X)3-7 (A/S/D)(W/V/A)(D/N)(V/A/Y)(F/V/I)(L/Q/V)(T/V/L)(V/C/I) DSDTX(L/I)(D/E/Y)(P/E/K)X(A/S/C)XXE (SEQ ID NO:
- 11);
- (ii) (L/M)(S/T/V)(D/S/A)XRY(W/Y/D)XAF(G/N/C) (V/I)ER(A/S)(C/A)QSX(F/T)X(N/C/T)(V/I)(L/Q/S)(C/V) (I/V/C)(S/G)GPL(G/S)XY(K/R) (SEQ ID NO:12);
- (iii) (D/E)X(W/Y)(X)2QXF(L/M)G(X)2(C/V)(S/T) XGDDR(H/R/C)LTN (SEQ ID NO:13);
- and
- (iv) (E/D)(T/S/V)P(X)5(W/Y/F)(L/I)XQQ(T/N) RW(S/T/N)KS(Y/W/F)(F/C)RE (SEQ ID NO:14),
- with X being selected from the group consisting of any amino acid;
- and
- wherein the amino acid motifs are found in the relative order of (i), (ii), (iii), and (iv) from an amino terminus to a carboxyl terminus of the enzymatically active hyaluronan synthase;
- and
- culturing the Bacillus strain under conditions appropriate for the production of hyaluronic acid.
9 more elements
A method of producing hyaluronic acid, comprising the steps of:
- introducing a purified nucleic acid segment having a coding region encoding enzymatically active hyaluronan synthase into a Bacillus strain, wherein the coding region encodes an enzymatically active hyaluronan synthase that includes at least two amino acid motifs selected from the group consisting of:
- (i) GKR(X)6F(X)11-15DSDT(X)8E (SEQ ID NO:15);
- (ii) RY(X)2AF(X)2ER(X)2QS(X)9GPL(X)2Y (SEQ ID NO:16);
- (iii) QXFXG(X)5GDDRXLTN (SEQ ID NO:17);
- and
- (iv) P(X)8QQXRWXKS(X)2RE (SEQ ID NO:18),
- with X being selected from the group consisting of any amino acid;
- and
- wherein the at least two amino acid motifs are found in the relative order of (i), (ii), (iii), and (iv) from an amino terminus to a carboxyl terminus of the enzymatically active hyaluronan synthase;
- and
- culturing the Bacillus strain under conditions appropriate for the production of hyaluronic acid.
8 more elements
A method of producing hyaluronic acid, comprising the steps of:
- introducing a purified nucleic acid segment having a coding region encoding enzymatically active hyaluronan synthase into a Bacillus strain, wherein the coding region encodes an enzymatically active hyaluronan synthase that includes the amino acid motifs:
- (i) GKR(X)6F(X)11-15DSDT(X)8E (SEQ ID NO:15);
- (ii) RY(X)2AF(X)2ER(X)2QS(X)9GPL(X)2Y (SEQ ID NO:16);
- (iii) QXFXG(X)5GDDRXLTN (SEQ ID NO:17);
- and
- (iv) P(X)8QQXRWXKS(X)2RE (SEQ ID NO:18),
- with X being selected from the group consisting of any amino acid;
- and
- wherein the amino acid motifs are found in the relative order of (i), (ii), (iii), and (iv) from an amino terminus to a carboxyl terminus of the enzymatically active hyaluronan synthase;
- and
- culturing the Bacillus strain under conditions appropriate for the production of hyaluronic acid.
8 more elements
Dependencies are inferred from the claim text.