US4337313A
Expired / Lapsed
US
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Immobilization of biocatalysts
Assignee: Haarmann and Reimer Corp
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Filing: 1980-12-08
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Granted: 1982-06-29
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Expires: 2000-12-08
Abstract
Abstract Enzymes or enzyme producing microorganisms are immobilized by contacting an aqueous medium containing an enzyme or enzyme producing microorganism with tannin, a long chain, polyamine, cationic flocculating agent and a crosslinking agent and separating the resultant enzymatically active reaction product from the aqueous medium. The reaction product has improved physical strength for use in a column bed reactor. A preferred flocculating agent is a water soluble epihalohydrin/polyamine copolymer.
Inventors
- 👤 Donald F. Hershberger
- 👤 Moshe M. Sternberg
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 11:35
CPC Classifications
C; C12; C12P; C12P19/00; C12P19/24; C12N; C12N11/00; C12N11/02; C12P19/20
Claims Analysis
Total Claims: 19
Claim 1
Independent Claim
116 words
A method for the immobilization of an enzyme producing microorganism or the enzyme produced thereby which comprises forming a reaction product by the steps of: (a) contacting an aqueous medium containing the enzyme or a mass of bacterial cells having biocatalytic activity with tannin; a long chain, polyamine, cationic flocculating agent selected from the group consisting of poly(acrylamide), poly(ethyleneimine), poly(2-hydroxypropyl-1-N-methylammonium chloride), poly(2-hydroxypropyl-1,1-N-dimethylammonium chloride), poly[N-(dimethylaminomethyl)-acrylamide], poly(diallyldimethylammonium chloride), poly(N,N-dimethylaminoethyl methacrylate), poly[N-dimethylaminopropyl]-methacrylamide and a water soluble epihalohydrin/polyamine copolymer and a crosslinking agent selected from the group consisting of a multifunctional aldehyde, a multifunctional organic halide, a multifunctional anhydride, a multifunctional azo compound, a multifunctional isocyanate and a multifunctional isothiocyanate, and (b) separating the reaction product from the aqueous medium.
Claim 2
Independent Claim
116 words
A method for the immobilization of an enzyme producing microorganism or the enzyme produced thereby which comprises forming a reaction product by the steps of: (a) contacting an aqueous medium containing the enzyme or a mass of bacterial cells having biocatalytic activity with tannin; a long chain, polyamine, cationic flocculating agent selected from the group consisting of poly(acrylamide), poly(ethyleneimine), poly(2-hydroxypropyl-1-N-methylammonium chloride), poly(2-hydroxypropyl-1,1-N-dimethylammonium chloride), poly[N-(dimethylaminomethyl)-acrylamide], poly(diallyldimethylammonium chloride), poly(N,N-dimethylaminoethyl methacrylate), poly[N-dimethylaminopropyl]-methacrylamide and a water soluble epihalohydrin/polyamine copolymer and a crosslinking agent selected from the group consisting of a multifunctional aldehyde, a multifunctional organic halide, a multifunctional anhydride, a multifunctional azo compound, a multifunctional isocyanate and a multifunctional isothiocyanate, and (b) separating the reaction product from the aqueous medium.
Claim 3
Dependent Claim
13 words
The method of claim 1 wherein the immobilized material is a free enzyme.
Claim 4
Dependent Claim
20 words
The method of claim 1 wherein the immobilized material is in the form of a whole or ruptured microorganism cell.
Claim 5
Dependent Claim
144 words
The method of claim 1 wherein the flocculating agent is water soluble epihalohydrin/polyamine copolymer obtained by the polymerization of an epihalohydrin with an alkylene polyamine having the formula R 1 R 2 NRNH 2 wherein R is a lower alkylene having from 2 to about 6 carbon atoms, and R 1 and R 2 are each lower alkyl of from 1 to about 6 carbon atoms, the mole ratio of epihalohydrin to polyamine being from about 0.60:1 to about 2.7:1, said polymerization comprising reacting with the alkylene polyamine from about 50 to about 90 percent of the amount of epihalohydrin to be polymerized, allowing the reaction to continue until the reaction medium attains a substantially uniform viscosity, and reacting the remaining portion of the epihalohydrin incrementally to obtain the cationic polymer, the temperature of polymerization being from about 60° C. to about 120° C.
Claim 6
Dependent Claim
11 words
The method of claim 1 wherein the crosslinking agent is glutaraldehyde.
Claim 7
Dependent Claim
10 words
The method of claim 1 wherein quebracho tannin is employed.
Claim 8
Dependent Claim
10 words
The method of claim 1 wherein tannic acid is employed.
Claim 9
Dependent Claim
25 words
The method of claim 1 wherein the cationic flocculating agent and crosslinking agent are prereacted to form an adduct before their contacting with the biocatalyst.
Claim 10
Dependent Claim
36 words
The method of claim 1 wherein the enzyme producing microorganism is Streptomyces olivaceous, S. griseus, S. olivochromogenes, S. phaeochromogenes, Bacillus licheniformis, B. amyloliquefaciens, B. coagulans, B. awamori, B. subtilis, Aspergillus niger, A. oryzae or Protaminobacter rubrum.
Claim 11
Independent Claim
71 words
A method for the immobilization of Streptomyces olivaceous which comprises the consecutive steps of: (a) providing an aqueous medium containing whole cells of the microorganism; (b) adding quebracho tannin to the aqueous medium; (c) adding an adduct of glutaraldehyde and an epihalohydrin/polyamine copolymer to the aqueous medium to form a reaction product; (d) removing the reaction product from the aqueous medium; and (e) drying the reaction product at an elevated temperature.
Claim 12
Independent Claim
71 words
A method for the immobilization of Streptomyces olivaceous which comprises the consecutive steps of: (a) providing an aqueous medium containing whole cells of the microorganism; (b) adding quebracho tannin to the aqueous medium; (c) adding an adduct of glutaraldehyde and an epihalohydrin/polyamine copolymer to the aqueous medium to form a reaction product; (d) removing the reaction product from the aqueous medium; and (e) drying the reaction product at an elevated temperature.
Claim 13
Independent Claim
66 words
A method for the immobilization of Bacillus licheniformis which comprises the consecutive steps of: (a) providing an aqueous medium containing the microorganism cells; (b) adding glutaraldehyde and an epihalohydrin/polyamine copolymer to the aqueous medium; (c) adding quebracho tannin to the aqueous medium to form a reaction product; (d) removing the reaction product from the aqueous medium; and (e) drying the reaction product at an elevated temperature.
Claim 14
Independent Claim
66 words
A method for the immobilization of Bacillus licheniformis which comprises the consecutive steps of: (a) providing an aqueous medium containing the microorganism cells; (b) adding glutaraldehyde and an epihalohydrin/polyamine copolymer to the aqueous medium; (c) adding quebracho tannin to the aqueous medium to form a reaction product; (d) removing the reaction product from the aqueous medium; and (e) drying the reaction product at an elevated temperature.
Claim 15
Independent Claim
63 words
A method for the immobilization of Protaminobacter rubrum which comprises the steps of: (a) providing an aqueous medium containing the microorganism cells; (b) adding tannin to the aqueous medium; (c) adding an adduct of glutaraldehyde and an epihalohydrin/polyamine copolymer to the aqueous medium to form a reaction product; (d) removing the reaction product from the aqueous medium; and (e) drying the reaction product.
Claim 16
Independent Claim
63 words
A method for the immobilization of Protaminobacter rubrum which comprises the steps of: (a) providing an aqueous medium containing the microorganism cells; (b) adding tannin to the aqueous medium; (c) adding an adduct of glutaraldehyde and an epihalohydrin/polyamine copolymer to the aqueous medium to form a reaction product; (d) removing the reaction product from the aqueous medium; and (e) drying the reaction product.
Claim 17
Dependent Claim
19 words
The method of claim 12 wherein the reaction product is contacted with sucrose to thereby convert it to palatinose.
Claim 18
Dependent Claim
22 words
The method of claim 12 wherein the tannin is quebracho tannin and it is added to the aqueous medium before the adduct.
Claim 19
Dependent Claim
14 words
An immobilized enzyme or enzyme producing microorganism produced by the method of claim 1.
References Cited (Backward Citations)
US3779869A
Cited
US3736231A
Cited
US4060456A
Cited
US3989596A
Cited
US3980521A
Cited
US4090919A
Cited
US4224411A
Cited
US4212943A
Cited
US4288552A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1985-06-28 | 1985-12-28 | 1986-06-29 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1989-06-28 | 1989-12-28 | 1990-06-29 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1993-06-28 | 1993-12-28 | 1994-06-29 | $7700.00 | Window Closed / Lapsed |