1. The lovastatin esterase enzyme immobilized on a solid support insoluble in water, characterized in that the enzyme is covalently bound to a solid support activated with an at least difunctional coupling reagent, the combination of solid support and at least difunctional coupling agent being such, that the immobilized lovastatin esterase exhibiting at least 5 times higher the hydrolytic activity towards lovastatin and salts thereof, in the presence of simvastatin andor salts thereof, than towards simvastatin and salts thereof.
2. (canceled)
3. (canceled)
4. The lovastatin esterase enzyme according to claim 1, characterized in that the solid support is a modified polysaccharide comprising di-(C1-6alkyl)amino-C1-6alkylcellulose, especially diethylaminoethylcellulose, and the at least difunctional reagent activating the solid support is cyanuric acid O-sulphonate or cyanuric halide, especially cyanuric chloride.
5. (canceled)
6. The lovastatin esterase enzyme according to claim 1, characterized in that the solid support is a modified silica gel, especially modified with amino-C1-6 alkyl-tri(C1-6 alkoxy)silane, especially an aminopropylsilanized silica gel, and the at least difunctional reagent activating the solid support is cyanuric acid O-sulphonate or cyanuric halide, especially cyanuric chloride.
7. The lovastatin esterase enzyme according to claim 1, characterized in that the solid support is a polygalactoside and the at least difunctional reagent activating the solid support is a compound of the formula
wherein Y represents \u2014SO2\u2014 or \u2014SO2\u2014(CHR)n\u2014SO2\u2014, where n represents an integer of from 1 to 18, and R represents a hydrogen atom or C1-6 alkyl, or Y represents \u2014SO2\u2014Ar\u2014SO2\u2014, where Ar represents a divalent aryl radical formed by displacing two hydrogen atoms directly bound to the aromatic ring carbon atoms, the divalent aryl radical optionally bearing C1-6 alkyl substituents.
8-10. (canceled)
11. The lovastatin esterase enzyme according to claim 1, characterized in that the solid support is a polygalactoside, and the at least difunctional reagent activating the solid support is the compound of the formula
wherein Y represents \u2014SO2\u2014.
12. The lovastatin esterase enzyme according to claim 1, characterized in that the solid support is diethylaminoethylcellulose, and the at least difunctional reagent activating the solid support is cyanuric chloride.
13. The lovastatin esterase enzyme according to claim 1, characterized in that the solid support is an aminopropylsilanized silica gel, and the at least difunctional reagent activating the solid support is cyanuric chloride.
14. The lovastatin esterase enzyme according to claim 1, characterized in that the enzyme is an enzyme produced by Clonostachys compactiuscula ATTC 38009, ATCC 74178.
15. A process for immobilization of the lovastatin esterase enzyme on a solid support insoluble in water, characterized in that using mechanical agitation, a cyanuric halide is contacted with a solid support comprising modified polysaccharide or modified silica gel in a solvent, the activated solid support is separated by filtration, the activated solid support is dried and suspended in an aqueous mixture containing the lovastatin esterase enzyme, until immobilization of the enzyme, the suspended material is separated by filtration, washed with a buffer and dried.
16. (canceled)
17. A process according to claim 15, characterized in that, the modified polysaccharide is a di-(C1-6alkyl)amino-C1-6alkylcellulose, especially diethylaminoethylcellulose.
18. (canceled)
19. A process according to claim 18, characterized in that a modified silica gel is silica gel modified with amino-C1-6alkyl-tri(C1-6alkoxy)silane, especially an aminopropylsilanized silica gel.
20. A process according to claim 15, characterized in that the cyanuric halide used is cyanuric chloride.
21. A process according to claim 15, characterized in that an autoclaved solid support is used.
22-24. (canceled)
25. A process according to claim 15, characterized in that the enzyme-containing aqueous solution used is a protein fraction of the material extracted from Clonostachys compactiuscula ATTC 38009, ATCC 74178.
26. A process for immobilization of the lovastatin esterase enzyme on a solid support insoluble in water, characterized in that the compound of the formula
wherein Y represents \u2014SO2\u2014 or \u2014SO2\u2014(CHR)n\u2014SO2\u2014, where n represents an integer of from 1 to 18, and R represents a hydrogen atom or C1-6 alkyl, or Y represents \u2014SO2\u2014Ar\u2014SO2\u2014, where Ar represents a divalent aryl radical formed by displacing two hydrogen atoms directly bound to the aromatic ring carbon atoms, the divalent aryl radical optionally bearing C1-6 alkyl substituents, is contacted with the solid polygalactose support in a solvent using mechanical agitation, the activated solid support is separated by filtration, the activated solid support is dried and suspended in an aqueous mixture containing the lovastatin esterase enzyme, the suspended material is separated by filtration, washed with a buffer and dried.
27. A process according to claim 26, characterized in that the compound of the formula
is a compound wherein Y represents \u2014SO2\u2014.
28. (canceled)
29. (canceled)
30. A process according to claim 26, characterized in that the enzyme-containing aqueous solution used is a protein fraction of the material extracted from Clonostachys compactiuscula ATTC 38009, ATCC 74178.
31-36. (canceled)
37. A biocatalytic flow reactor with a bed comprising a body of the reactor with an inner space connected to the fluid inlet and connected to the fluid outlet, in which inner space there is a bed containing the lovastatin esterase enzyme immobilized on a solid support insoluble in water, characterized in that the enzyme is covalently bound to the solid support activated with an at least difunctional coupling reagent, the combination of solid support and at least difunctional coupling agent being such, that the immobilized lovastatin esterase exhibits at least 5 times higher the hydrolytic activity towards lovastatin and salts thereof in the presence of simvastatin andor salts thereof, than towards simvastatin and salts thereof.
38-44. (canceled)
45. A biocatalytic flow reactor according to claim 37, characterized in that the solid support is a polygalactoside, and the at least difunctional reagent activating the solid support is a compound of the formula
wherein Y represents \u2014SO2\u2014.
46. A biocatalytic flow reactor according to claim 37, characterized in that the solid support is diethylaminoethylcellulose, and the at least difunctional reagent activating the solid support is cyanuric chloride.
47. A biocatalytic flow reactor according to claim 37, characterized in that the enzyme is an enzyme produced by Clonostachys compactiuscula ATTC 38009, ATCC 74178.
48. A process for preparation andor purification of simvastatin comprising treating the solution of the simvastatin salt containing residual content of the lovastatin salt with the lovastatin esterase enzyme until hydrolysing lovastatin to form the triol, separating the triol, and isolating simvastatin substantially free from lovastatin, where the solution of the simvastatin salt containing residual content of the lovastatin salt is brought into a contact with the lovastatin esterase enzyme immobilized on a solid support insoluble in water, characterized in that the enzyme is covalently bound to the solid support activated with an at least difunctional coupling reagent, the combination of solid support and at least difunctional coupling agent being such, that the immobilized lovastatin esterase exhibits at least 5 times higher the hydrolytic activity towards lovastatin and salts thereof in the presence of simvastatin andor salts thereof, than towards simvastin and salts thereof.
49-58. (canceled)
59. A process for preparation andor purification of simvastatin according to claim 48, characterized in that the solid support is a polygalactoside, and the at least difunctional reagent activating the solid support is the compound of the formula
wherein Y represents \u2014SO2\u2014.
60. A process for preparation andor purification of simvastatin according to claim 48, characterized in that the solid support is diethylaminoethylcellulose, and the at least difunctional reagent activating the solid support is cyanuric chloride.
61. A process for preparation andor purification of simvastatin according to claim 48, characterized in that the solid support is an aminopropylsilanized silica gel, and the at least difunctional reagent activating the solid support is cyanuric chloride.
62. (canceled)
63. (canceled)
64. A process for preparation andor purification of simvastatin according to claim 48, characterized in that the enzyme is an enzyme produced by Clonostachys compactiuscula ATTC 38009, ATCC 74178.
65. The lovastatin esterase enzyme according to claim 11, characterized in that the enzyme is an enzyme produced by Clonostachys compactiuscula ATTC 38009, ATCC 74178.
66. The lovastatin esterase enzyme according to claim 12, characterized in that the enzyme is an enzyme produced by Clonostachys compactiuscula ATTC 38009, ATCC 74178.
67. The lovastatin esterase enzyme according to claim 13, characterized in that the enzyme is an enzyme produced by Clonostachys compactiuscula ATTC 38009, ATCC 74178.
68. A process according to claim 17, characterized in that an autoclaved solid support is used.
69. A process according to claim 19, characterized in that an autoclaved solid support is used.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. A pallet pulling device for moving a pallet to position for engagement by the tines of a forklift truck, the pallet having normally horizontal planks and a stringer secured to and extending vertically about the planks, the pallet pulling device comprising:
a. a puller bar having an upper face, a lower face, and side edge surfaces, and,
b. a crossbar secured transversely to the puller bar to define a generally T-shaped pulling device, the crossbar having an upper face, a lower face and opposite side edge surfaces, the upper face of the crossbar being in facial engagement with the lower face of the puller bar,
c. the pulling device being positively engageable with a pallet with the lower face of the puller bar engaging one of said planks, the crossbar extending under a lower edge of a finger, a side edge of the puller bar engaging a side face of the stringer, and a side edge of the crossbar engaging a side edge of a plank.
2. A pallet pulling device according to claim 1, wherein said crossbar is attached to the puller bar at such a position that the crossbar has a first relatively short bar portion extending in one direction from the puller bar and a second relatively long bar portion extending oppositely from the puller bar.
3. A pallet pulling device according to claim 2, wherein said second relatively long crossbar portion is about twice the length of the first relatively short bar.
4. A pallet pulling device according to claim 1, wherein the puller bar and the crossbar are of generally rectangular cross-section, and the crossbar and puller bar are oriented generally horizontally when the pulling device engages the pallet.
5. A method of pulling a pallet comprising the steps of:
a. fashioning a flat puller bar;
b. welding a crossbar to a bottom tip of the flat puller bar, wherein the crossbar is welded at an inside weld, an outside weld, and a top weld, whereby a pallet pulling device is formed;
c. placing the pallet pulling device so that the crossbar fits under a pallet stringer of the pallet, wherein the flat puller bar is parallel to the pallet stringer, wherein the crossbar pulls against the rear edge of a plank of the pallet
d. applying pulling force to the flat puller bar to pull the pallet.
6. The method of claim 5, wherein the crossbar comprises a short bar portion and a long bar portion, wherein the long bar portion is approximately twice the length of the short bar portion.
7. The method of claim 5 further comprising the step of forming a hole on the flat puller bar opposite the top weld.
8. The method of claim 5 wherein the flat puller bar is perpendicular to the crossbar.
9. The method of claim 5 wherein the flat puller bar has a rectangular cross-section.
10. The method of claim 5 wherein the crossbar has a rectangular cross-section.
11. The method of claim 10 wherein the flat puller bar has a rectangular cross-section.
12. The method of claim 10 wherein the crossbar extends under the edge of a stringer of the pallet to prevent disengagement of the pulling device from the pallet when pulling force is applied, wherein a hole is located on the flat puller bar; an outside weld opposes an inside weld, wherein both welds are adjacent to a top weld that is in opposition to the location of the hole.