1.-3. (canceled)
4. An isolated cyclodipeptide synthetase, which:
a) has an ability to produce cyclo(Leu-Leu) cyclodipeptide from two Leu amino acids, and
b) comprises a polypeptide sequence having at least 34% of identity or at least 56% of similarity, with at least one of the polypeptides selected from the group consisting of sequences SEQ ID No 2, SEQ ID No 4, SEQ ID No 6 and SEQ ID No 8.
5.-6. (canceled)
7. An isolated polynucleotide selected from the group consisting of:
a) a polynucleotide encoding a cyclodipeptide synthetase as defined in claim 4;
b) a complementary polynucleotide of the polynucleotide a);
c) a polynucleotide which hybridises to polynucleotide a) or b) under stringent hybridization conditions.
8. The isolated polynucleotide according to claim 7, which is selected from the group consisting of the polynucleotides of sequences SEQ ID No 1, SEQ ID No 3, SEQ ID No 5 and SEQ ID No 7.
9. A recombinant vector comprising a polynucleotide as defined in claim 6.
10. A modified host cell comprising a polynucleotide as defined in claim 6.
11.-12. (canceled)
13. A method for the synthesis of a cyclo(Leu-Leu) cyclodipeptide, which comprises the steps of:
(1) incubating leucine, under suitable conditions, with at least one cyclodipeptide synthetase as defined in claim 4, and
(2) recovering the cyclo(Leu-Leu) cyclodipeptide thus obtained.
14. A method for the synthesis of \u03b1,\u03b2-dehydrogenated cyclo (Leu-Leu) cyclodipeptide, which comprises the steps of:
(1) incubating leucine, under suitable conditions, with a cyclodipeptide synthetase enzyme as defined in claim 4, and a purified CDO, and
(2) recovering the \u03b1,\u03b2-dehydrogenated cyclodipeptide.
15. The method according to claim 13, wherein step (1) is performed in presence of Leu at a concentration between 0.1 mM and 100 mM, said cyclodipeptide synthetase at a concentration between 0.1 mM and 100 \u03bcM, in a buffer at pH of between 6 and 8, and containing a soluble extract of prokaryote cells such as E. coli or Streptomyces cells which does not produce cyclodipeptide synthetase.
16. The method according to claim 14, which further comprises, prior or simultaneously with step (1), a step synthesizing said cyclodipeptide synthetase using a polynucleotide as defined in claim 7.
17. A method for producing cyclo(Leu-Leu) cyclodipeptide which comprises the steps of:
(1) culturing a host cell according to claim 10, in suitable culture conditions for said host cell, and
(2) recovering the cyclo(Leu-Leu) cyclodipeptide from the culture medium.
18. A method for the synthesis of \u03b1,\u03b2-dehydrogenated cyclo(Leu-Leu) cyclodipeptide, which comprises the following steps:
(1) culturing a modified host cell according to claim 10 in appropriate culture conditions for said host cell,
(2) incubating the cyclo(Leu-Leu) cyclodipeptide obtained from step (1\u2032) with a purified CDO, and
(3) recovering the \u03b1,\u03b2-dehydrogenated cyclo(Leu-Leu) cyclodipeptide derivative from the culture medium.
19. \u03b1,\u03b2-dehydrogenated cyclo(Leu-Leu) cyclodipeptides, which are selected from the group consisting of cyclo(\u0394Leu-Leu)\xb1cyclo(\u0394Leu-\u0394Leu), and a mixture thereof.
20. The method according to claim 15, wherein said Leu is in a concentration of 1 mM to 10 mM.
21. The method according to claim 15, wherein said cyclodipeptide synthetase is in a concentration of 1 \u03bcM to 100 \u03bcM.
22. The method according to claim 15, wherein said prokaryote cells are E. coli.
23. The method according to claim 15, wherein said prokaryote cells are Streptomyces cells.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
What is claimed is:
1. An apparatus for improving a golf stroke along a line of intended travel of a golf ball by aligning a user’s eyes at an eye location point above the apparatus within a vertical plane that includes the golf ball and the line of intended travel, comprising:
a. a frame structure, comprising a first support leg and a second support leg spaced from the first support leg, and
b. an eye alignment device supported by the frame structure, comprising:
i. a substantially linear top edge portion that is visible when viewed from the eye location point, and
ii. a tail portion spaced from the top edge portion and located within the vertical plane,
wherein the tail portion is substantially obscured from view by the top edge portion when the user’s eyes are located within the vertical plane.
2. The apparatus of claim 1, wherein the eye alignment device further comprises a plurality of substantially linear indicators, each indicator being aligned within the vertical plane that includes the golf ball and the line of intended travel.
3. The apparatus of claim 2, wherein the eye alignment device comprises two spaced and substantially linear indicators.
4. The apparatus of claim 1, wherein the top edge portion comprises a first substantially linear indicator and wherein the tail portion comprises a second substantially linear indicator.
5. The apparatus of claim 4, wherein the first indicator comprises a first elastic portion and wherein the second indicator comprises a second elastic portion.
6. The apparatus of claim 1, wherein the first support leg comprises a first arm and the second support leg comprises a second arm, and wherein the eye alignment device comprises a continuous elastic loop passed around the respective first and second arms to form an upper course and a lower course of the elastic loop, so that the top edge portion comprises the upper course and the tail portion comprises the lower course of the elastic loop.
7. An apparatus for improving a golf stroke, for example a putting stroke, by aligning a heel portion of a putter along a desired stroke plane that includes a horizontal line that is parallel to a line of intended travel of a golf ball along the ground, comprising:
a. a frame structure, comprising a first support leg and a second support leg spaced from the first support leg, and
b. a putter alignment member defining
i. a first end affixed to the first support leg,
ii. a spaced second end affixed to the second support leg, and
iii. a substantially planar contact face,
wherein the contact face is oriented so that a horizontal line extending along the contact face is parallel to the line of intended travel, and so that when the heel portion of the putter head is moved back and forth in the putting stroke with the heel portion in contact with the contact face, the heel portion moves within a plane that is parallel to the contact face.
8. The apparatus of claim 7, wherein the contact face defines a substantially vertical plane.
9. The apparatus of claim 7, wherein the contact face defines a plane that is inclined with respect to the ground, so that so that an angle measured between the contact face and the ground along a vertical plane perpendicular to the line of intended travel of the golf ball is in a range of from about 90 degrees to about 180 degrees.
10. The apparatus of claim 9, wherein the contact face defines an angle measured between the contact face and the ground along a vertical plane perpendicular to the line of intended travel of the golf ball is in a range of from about 95 degrees to about 120 degrees.
11. The apparatus of claim 9, wherein the contact face defines an angle measured between the contact face and the ground along a vertical plane perpendicular to the line of intended travel of the golf ball of about 107 degrees.
12. The apparatus of claim 7, further comprising an adjustable coupling being adapted for selectively connecting the putter alignment member to the frame structure so that so that an angle measured between the contact face and the ground along a vertical plane perpendicular to the line of intended travel of the golf ball is in a range of from about 90 degrees to about 180 degrees.
13. The apparatus of claim 12, wherein the putter alignment member further defines a bottom edge adjacent the ground, a top edge spaced from the bottom edge, a first side edge between the top edge and the bottom edge, and a second side edge between the top edge and the bottom edge and spaced from the first side edge.
14. The apparatus of claim 13, wherein the adjustable coupling comprises
a. a first lower stud projecting from the first side edge of the putter alignment member adjacent the bottom edge,
b. a second lower stud projecting from the second side edge adjacent the bottom edge,
c. a first upper stud projecting from the first side edge and spaced above the first lower stud, and
d. a second upper stud projecting from the second side edge and spaced above the second lower stud,
wherein the first support leg defines,
e. a first hole sized and shaped to receive the first lower stud therein, and
f. a first slot sized and shaped to receive the first upper stud therein and spaced from the first hole,
wherein the second support leg defines,
g. a second hole sized and shaped to receive the second lower stud therein, and
h. a second slot sized and shaped to receive the second upper stud therein and spaced from the second hole,
wherein the respective studs and slots allow for rotational adjustment of the contact face about an axis defined by a line extending between the first lower stud and the second lower stud.
15. The apparatus of claim 14, wherein the adjustable coupling further comprises a plurality of retainers, one respective retainer engaging each of the first lower stud, the second lower stud, the first upper stud and the second upper stud, for selectively securing the putter alignment member in engagement with the frame structure.
16. An apparatus for improving a golf stroke by allowing a golfer to align their feet with an intended line of travel of a golf ball along the ground, comprising:
a. a frame structure, comprising a first support leg and a second support leg spaced from the first support leg, and
b. a stance alignment device supportable by the frame structure, comprising a substantially linear foot reference indicator extending between the first support leg and the second support leg, the foot reference indicator being substantially parallel to the line of intended travel of the golf ball.
17. The apparatus of claim 16, further comprising at least one connector for connecting the stance alignment device to the frame structure.
18. The apparatus of claim 16, wherein the stance alignment device further comprises at least one marker for indicating a selected position for the golfer’s feet during the golf stroke.
19. The apparatus of claim 18, wherein the stance alignment device further comprises a first adjustable marker for indicating a selected position for the golfer’s right foot, and a second adjustable marker for indicating a selected position for the golfer’s left foot during the golf stroke.
20. The apparatus of claim 16 for further improving the golf stroke by aligning a heel portion of a putter along a desired stroke plane that includes the line of intended travel of the golf ball, further comprising:
a. a putter alignment member supported by the frame structure, defining a substantially planar contact face oriented so that a horizontal line along the contact face is parallel to the line of intended travel, so that when the heel portion of the putter head is moved back and forth in a putting motion with the heel portion in contact with the contact face, the heel portion moves within a plane that is parallel to the contact face.
21. The apparatus of claim 16, wherein the first support leg comprises a first base member and the second support leg comprises a second base member, the stance alignment device being affixed to the respective first base member and second base member.
22. The apparatus of claim 21, wherein the stance alignment device comprises an elastic member having a first end and a spaced second end, the first end engaging the first base member and the second end engaging the second base member.
23. The apparatus of claim 22, wherein the stance alignment device further comprises:
a. a first connector for connecting the first end of the elastic member to the first base member, and
b. a second connector for connecting the second end of the elastic member to the second base member.
24. The apparatus of claim 23, wherein the first base member comprises a portion defining at least one aperture therethrough, the at least one aperture being sized and shaped to receive the first connector therein, and wherein the second base member comprises a portion defining at least one aperture therethrough, the at least one aperture being sized and shaped to receive the second connector therein.
25. An apparatus for improving a golf stroke by imparting a force to a putter, the putter having a putter head and a connection location, tending to maintain the putter head along a line of intended travel of a golf ball when the putter head is moved back and forth in a putting motion, comprising:
a. a frame structure, comprising a first support leg having a first base member, and a second support leg spaced from the first support leg and having a second base member, and
b. a stroke resistance device supported by the frame structure, comprising
i. a first resistance component, having a fixed end and an variable end spaced from the fixed end, the fixed end being connected to the first base member at a first fixed point and the variable end being connected to the putter at the connection location, and
ii. a second resistance component, having a fixed end and a variable end spaced from the fixed end, the fixed end being connected to the second base member at a second fixed point and the variable end being connected to the putter at the connection location,
wherein, when the putter head is moved back and forth in the putting motion generally between the first fixed point and the second fixed point, at least one of the first or second resistance components imparts a tensile force to the putter head toward the respective first or second fixed points.
26. The apparatus of claim 25, wherein the first resistance component comprises a first elastic member, and wherein the second resistance component comprises a second elastic member.
27. The apparatus of claim 26, wherein the first resistance component further comprises a first connector for connecting the first elastic member to the first base member, and wherein the second resistance component further comprises a second connector for connecting the second elastic member to the second base member.
28. The apparatus of claim 27, wherein the first base member defines a plurality of apertures therethrough, each aperture sized and shaped to engage the first connector, and wherein the second base member defines a plurality of apertures therethrough, each aperture sized and shaped to engage the second connector.
29. An apparatus for improving a golf stroke along a line of intended travel of a golf ball by positioning a putter having a putter shaft connected to a putter head with a heel portion and a sole surface, so that the sole surface is substantially flat upon the ground, comprising:
a. a frame structure, comprising a first support leg and a second support leg spaced from the first support leg, and
b. a putter alignment member supported by the frame structure and defining a substantially planar contact face oriented so that a horizontal line along the contact face is substantially parallel to the line of intended travel,
c. a substantially linear shaft alignment device supported by the frame structure and oriented substantially parallel to the line of intended travel,
wherein the shaft alignment device is attached to the frame structure at a selected location so that when the heel portion of the putter head is in contact with the contact face, and the putter shaft is in contact with the shaft alignment device, the sole surface of the putter head is substantially flat upon the ground.
30. The apparatus of claim 29, wherein the first support leg comprises a first upright member, and wherein the second support leg comprises a second upright member, the respective first and second upright members engaging the shaft alignment device.
31. The apparatus of claim 30, wherein the first upright member defines a plurality of openings therethrough, and wherein the second base member defines a plurality of openings therethrough, each opening defined by the respective first and second upright members being sized and shaped to engage the shaft alignment device.
32. The apparatus of claim 31, wherein the shaft alignment device comprises a substantially cylindrical and elongated rod having a first end engaging the first upright member and a second end engaging the second upright member that is spaced from the first end.