Hyaluronan synthase gene and uses thereof
AssigneeThe Board of Regents of the University of Oklahoma
InventorsPaul Weigel, Kshama Kumari, Paul DeAngelis
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Abstract
The present invention relates to a recombinant host cell that includes an expression construct having a hyaluronan synthase gene that encodes an enzymatically active hyaluronan synthase (HAS), and to the use of these recombinant cells that produce hyaluronan synthase and its hyaluronic acid product.
Claims48
A recombinant host cell comprising at least one expression construct comprising a hyaluronan synthase gene, wherein the hyaluronan synthase gene encodes an enzymatically active hyaluronan synthase, wherein the enzymatically active hyaluronan synthase 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
- (SEQ ID NO:11)
- (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;
- (ii)
- (L/M)(S/T/V)(D/S/A)XRY(W/Y/D)XAF(G/N/C)
- (SEQ ID NO:
- 12)
- (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);
- (iii)
- (D/E)X(W/Y)(X)2QXF(L/M)G(X)2(C/V)(S/T)
- (SEQ ID NO:13)
- XGDDR(H/R/C)LTN;
- and
- (iv)
- (E/D)(T/S/V)P(X)5(W/Y/F)(L/I)XQQ(T/N)
- (SEQ ID NO:
- 14)
- RW(S/T/N)KS(Y/W/F)(F/C)RE,
- 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 recombinantly produced, enzymatically active hyaluronan synthase.
A recombinant host cell comprising at least one expression construct comprising a hyaluronan synthase gene, wherein the hyaluronan synthase gene encodes an enzymatically active hyaluronan synthase, wherein the enzymatically active hyaluronan synthase includes at least three 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
- (SEQ ID NO:11)
- (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;
- (ii)
- (L/M)(S/T/V)(D/S/A)XRY(W/Y/D)XAF(G/N/C)
- (SEQ ID NO:
- 12)
- (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);
- (iii)
- (D/E)X(W/Y)(X)2QXF(L/M)G(X)2(C/V)(S/T)
- (SEQ ID NO:13)
- XGDDR(H/R/C)LTN;
- and
- (iv)
- (E/D)(T/S/V)P(X)5(W/Y/F)(L/I)XQQ(T/N)
- (SEQ ID NO:
- 14)
- RW(S/T/N)KS(Y/W/F)(F/C)RE,
- with X being selected from the group consisting of any amino acid;
- and
- wherein the at least three 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 recombinantly produced, enzymatically active hyaluronan synthase.
A recombinant host cell comprising at least one expression construct comprising a hyaluronan synthase gene, wherein the hyaluronan synthase gene encodes an enzymatically active hyaluronan synthase, wherein the enzymatically active hyaluronan synthase 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
- (SEQ ID NO:11)
- (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;
- (ii)
- (L/M)(S/T/V)(D/S/A)XRY(W/Y/D)XAF(G/N/C)
- (SEQ ID NO:
- 12)
- (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);
- (iii)
- (D/E)X(W/Y)(X)2QXF(L/M)G(X)2(C/V)(S/T)
- (SEQ ID NO:13)
- XGDDR(H/R/C)LTN;
- and
- (iv)
- (E/D)(T/S/V)P(X)5(W/Y/F)(L/I)XQQ(T/N)
- (SEQ ID NO:
- 14)
- RW(S/T/N)KS(Y/W/F)(F/C)RE,
- 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 recombinantly produced, enzymatically active hyaluronan synthase.
A recombinant host cell comprising at least one expression construct comprising a hyaluronan synthase gene, wherein the hyaluronan synthase gene encodes an enzymatically active hyaluronan synthase, wherein the enzymatically active hyaluronan synthase 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 recombinantly produced, enzymatically active hyaluronan synthase.
A recombinant host cell comprising at least one expression construct comprising a hyaluronan synthase gene, wherein the hyaluronan synthase gene encodes an enzymatically active hyaluronan synthase, wherein the enzymatically active hyaluronan synthase includes at least three 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 three 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 recombinantly produced, enzymatically active hyaluronan synthase.
A recombinant host cell comprising at least one expression construct comprising a hyaluronan synthase gene, wherein the hyaluronan synthase gene encodes an enzymatically active hyaluronan synthase, wherein the enzymatically active hyaluronan synthase 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 recombinantly produced, enzymatically active hyaluronan synthase.
Dependencies are inferred from the claim text.