Claims for Patent: 4,321,327

Summary for Patent: 4,321,327

Title:	Preparation of spherical shaped mycelial pellets
Abstract:	Spherical shaped mycelial pellets having a support core and which are suitable for biocatalytic conversion of organic compounds are produced by dissolving a cellulose derivative in a solvent to form a solution, mixing spores of a mycelial microorganism with the solution, precipitating the cellulose derivative to form porous beads containing the spores, and incubating the beads in a culture medium. Alternatively, the pellets may be produced by dissolving agar in water, mixing the agar solution with spores of a mycelial microorganism, precipitating the agar to form beads containing the spores and incubating the beads in a culture medium.
Inventor(s):	Chen; Li F. (West Lafayette, IN), Gong; Cheng S. (West Lafayette, IN), Tsao; George T. (West Lafayette, IN)
Assignee:	Purdue Research Foundation (West Lafayette, IN)
Application Number:	06/100,816
Patent Claims:	1. A process for the preparation of biologically active mycelial pellets having a physical support core which comprises the steps of: (a) dissolving a hydrolyzable cellulose derivative on an inert organic, water-miscible solvent to form a solution having a density greater than that of a precipitation solution; (b) mixing spores of a mycelial microorganism with said solution and distributing the resulting solution in the form of droplets into a precipitation solution or cool air whereby said cellulose derivative is precipitated in the form of uniformly porous beads, said beads containing said spores; (c) separating the precipitated beads from said precipitation solution if used; (d) incubating the precipitated porous beads in an aqueous culture medium for a period of time sufficient to produce a round pellet characterized by a rigid spherical porous bead core surrounded by a porous integral webbed layer, said layer forming a spherical encasement of structural integrity about the rigid core and being composed of filamentous hyphae of a mycelial microorganism. 2. A process according to claim 1, wherein said cellulose derivative is selected from the group consisting of cellulose acetate, cellulose triacetate, and cellulose nitrate. 3. A process according to claim 1 wherein said microorganism is a fungi selected from the genus Rhizopus or Mucor. 4. A process according to claim 1 wherein the precipitated porous beads are washed with sterile water prior to incubating. 5. A process according to claim 4 wherein the incubation is carried out with agitation. 6. A process according to claim 1 wherein the distributing is accomplished by spraying. 7. A process according to claim 1 wherein said precipitation solution is selected from the group consisting of water, hexane, cyclohexane, octane, benzene and mixtures of water and ethanol or methanol. 8. A process according to claim 7 wherein said precipitation solution is water. 9. A process according to claim 1 wherein said solvent is a mixture of: (a) a member from the group consisting of acetone, a mixture of acetone and methanol or ethanol, methyl acetate, a mixture of methylene dichloride and methanol, methyl ethyl ketone, formamide and dimethyl sulfoxide; and (b) a member from the group consisting of dimethyl sulfoxide, formamide, methyl acetate, cyclohexanone, methylene dicholoride, ethylene dichloride, a mixture of methylene dicholoride and methanol, and a mixture of ethylene dicholoride and methanol. 10. A process according to claim 9 wherein said solvent is dimethyl sulfoxide, formamide or methyl acetate. 11. A process for the preparation of biologically active mycelial microorganism pellets which comprise the steps of: (a) dissolving agar in hot water to form an agar solution; (b) cooling the agar solution to a temperature of greater than 45.degree. C. but below the temperature at which the spores added in step (c) would be killed; (c) mixing the agar solution of step (b) with spores of a mycelial microorganism and dispersing the resulting mixture in the form of droplets into cool air or an aqueous precipitation solution at a temperature below 45.degree. C. whereby said agar is precipitated in the form of uniform beads containing said spores; (d) incubating the agar beads in an aqueous culture medium for a period of time sufficient to produce a round pellet characterized by a rigid round agar core surrounded by a porous integral webbed layer, said layer forming a spherical encasement of structural integrity about the rigid core and being composed of filamentous hyphae of a mycelial microorganism. 12. A process according to claim 11 wherein the mycelial spores are fungi of the genus Rhizopus or Mucor. 13. A process according to claim 11 wherein the agar beads are washed with sterile water prior to incubating. 14. A process according to claim 11 or claim 12 wherein dispersing the said mixture is accomplished by spraying. 15. A process according to claim 11 wherein the cooling in step (b) is carried out at a temperature ranging between about 45.degree. to 80.degree. C. 16. A process according to claim 15 wherein the medium solution is cooled to about 50.degree. C. 17. A process according to claim 11 wherein the aqueous precipitation solution is the aqueous culture medium. 18. A process according to claim 11 wherein the droplets are dispersed by spraying into cool air. 19. A process according to claim 11 wherein the droplets are dispersed by spraying into cool water. 20. A spherical shaped mycelial pellet produced according to the process of claim 1. 21. A pellet according to claim 20 wherein said cellulose derivative is selected from the group consisting of cellulose acetate, cellulose triacetate and cellulose nitrate. 22. A spherical shaped mycelial pellet produced according to the process of claim 11. 23. A pellet according to claim 1 or claim 22 wherein said microorganism is a fungi of the genus Rhizopus, Mucor, Aspergillus, Pencillium or Trichoderma. 24. A pellet according to claim 20 or claim 22 having a core diameter ranging from about 0.1 mm to 1.0 cm and a layer thickness of about 1 mm to 5 mm. 25. A method for the biocatalytic conversion of organic compounds which comprises reacting said compounds in the presence of the mycelial pellet of claim 20 or claim 22. 26. A method according to claim 25 wherein a carbohydrate material is converted to alcohol and said microorganism is a fungi of the genus Rhizopus, Mucor, or mixtures thereof. 27. A method according to claim 25 wherein said conversion is carried out continuously in a column reactor containing said mycelial pellets. 28. A biocatalytic column reactor containing the mycelial pellet of claim 20 or claim 22.

Details for Patent 4,321,327

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Sharp & Dohme Corp.	ZOSTAVAX	zoster vaccine live	For Injection	125123	05/25/2006		2039-09-23