enzymes and enzyme systems

Enzymes and Enzyme Systems

5128251

I M M O B I L I Z E D L I P O L Y T I C E N Z Y M E F O R E S T E R I F I C A T I O N

A N D I N T E R E S T E R I F I C A T I O N

Hideki Yokomichi, Takeshi Yasumasu, Kazuhir Nakamura, Yoshiharu Kawahara, Ibaraki, Japan assigned to Kao Corporation

An immobilized lipolytic enzyme for esterifica- tion and interesterification of fats and oils is prepared by adsorbing a fatty acid or a derivative thereof onto an insoluble carrier and then adsorbing a lipolytic enzyme onto the carrier in an aqueous medium. The resultant immobilized lipolytic enzyme can be dried to a water content of 2 wt. ~ or less. Preferably, the carrier is a phenol/formaldehyde resin, the lipolytic enzyme is from Rhizopus japonicus and the fatty acid is linoleic acid, lauric acid, stearic acid, ricinoleic acid or isostearic acid. A reaction column can be charged with the dried immobilized lipolytic enzyme and reaction continuously carried out without the use of a solvent. Preferably, starting materials have a water content of 0.01 to 0.2 wt. % and residence time in the column is 2 hours or shorter.

5128263

E N Z Y M A T I C R E S O L U T I O N P R O C E S S H Y D R O L Y Z I N G

T H I O - E S T E R

Jeffrey M Howell, Ramesh Patel, Laszlo J Szarka assigned to E R Squibb & Sons Inc

A novel enzymatic resolution process for pre- paring resolved compounds of the formula See

649

Patent for Chemical Structure I' with improved yields and high optical purity is disclosed. The thioester bond of the compound is hydrolyzed and the hydrolyzed compound is separated from the unhydrolyzed. Compounds of formula I' are useful, for example, as intermediates for the preparation of physiologically active compounds, e.g. captopril and zofenopril.

5128339

P R O T E O L Y T I C E N Z Y M E I N H I B I T I O N M E T H O D

Richard P Dunlap, Neil W Boaz, Albert J Mura, Virendra Kumar, Chakrapani Subramanyam, Ranjit C Desai, Dennis J Hlasta, Manohar T Saindane, Malcolm Bell, John J Court assigned to Sterling Winthrop Inc

4-R4-R5-2-Saccharinylmethyl aryl carbox- ylates, useful in the treatment of degenerative diseases, are prepared by reacting a 4-R4-R5-2- halomethylsaccharin with an arylcarboxylic acid in the presence of an acid-acceptor.

5118404

E N Z Y M E E L E C T R O D E A N D A M E T H O D O F D E T E R M I N I N G

C O N C E N T R A T I O N O F A N A N A L Y T E I N A S A M P L E

S O L U T I O N

Atsushi Saito, Tokyo, Japan assigned to NEC Corporation

An enzyme electrode usable for determining

650 PATENT ABSTRACTS

high concentration of analyte in a sample solution is provided, by providing an enzyme- immobilized membrane in the electrode with pH buffer capacity. Durability of the electrode can be improved by using albumin crosslinked by glutaraldehyde as a permeation-restricted membrane in the electrode. Method of using the electrode can be simplified by introducing a stirring step, measuring outputs before and after the stirring step and utilizing difference of measured outputs.

5118623

B L E A C H S T A B L E E N Z Y M E S

Georg Boguslawski, John W Shultz assigned to Solvay Enzymes lnc

The methods of the invention can be used to identify sites of cleavage of an enzyme in the presence of hypochlorite. It has been found that such cleavage occurs at tryptophan. Chemical modifications or genetic manipulation to change or delete tryptophan can be done to produce a more stable enzyme which retains activity in the presence of hypochlorite. The invention is particularly applicable to alkaline proteases which are useful in detergent compositions.

5120651

R E S T R I C T I O N E N Z Y M E A G E I A N D P R O C E S S F O R P R O D U C I N G

S A M E

novel compositions for enzyme complementa- tion assays for qualitative and quantitative determination of a suspected analyte in a sample. The use of enzyme-acceptor and enzyme-donor polypeptides prepared by recombinant DNA techniques, DNA synthesis or chemical poly- peptide synthesis techniques which are capable of interacting to form an active enzyme complex having catalytic activity characteristic of beta- galactosidase is described. Both homogeneous and heterogeneous assays utilizing these polypeptides are described.

5120837

D N A E N C O D I N G P H E A F E E D B A C K I N H I B I T I O N

R E S I S T A N T E N Z Y M E A N A L O G U E S

lan Fotheringham, Jennifer Nelms assigned to The NutraSweet Company

Disclosed are DNA sequences bonding deletion, substitution and/or addition analogs of the E. coli enzyme, chorismate mutase/prephenate dehydratase (CMPD). Preferred expression products include (des-Gin307, des-Ala308, des- Gly309, des-Ala310)CMPD; (Leu306)CMPD; (des-Thr304, Lys305, des-GIn306)CMPD; and (Cys309)CMPD display enzymatic activity of the wild type enzyme but are more resistant to inhibition in the presence of phenylalanine.

5122452

Yuzo Yamada, Hirofumi Mizuno, Kazuhid Yamasato, Fujieda, Japan assigned to Nisshin Seito Kabushiki Kaisha

A restriction enzyme capable of recognizing and cleaving a DNA sequence at a position indicated by the arrows: See Patent for Tabular Presenta- tion PS See Patent for Tabular Presentation

5120653

V E C T O R C O M P R I S I N G D N A S E Q U E N C E C O D I N G F O R

E N Z Y M E - D O N O R P O L Y P E P T I D E

Daniel R Henderson assigned to Microgenics Corporation

This invention relates to improved methods and

E N Z Y M E I M M U N O A S S A Y W I T H A M A C R O P O R O U S

H Y D R O P H O B I C S Y N T H E T I C P O L Y M E R C L O T H C O N T A I N I N G

A N I M M O B I L I Z E D A N T I B O D Y O R A N T I G E N

Hiroshi Yamazaki, Burton W Blais, Nepean, Canada assigned to Carleton University

An immunoassay device containing an immobilized antibody or antigen is provided by directly absorbing and absorbing an unmodified antibody or antigen on and within a woven or non-woven macroporous hydrophobic synthetic polymer cloth formed of a synthetic polymer selected from the group consisting of plypropy- lene, polyester, nylon and polyethylene. The cloth has a thickness of more than about 200 mum and contains pores in the form of spaces

PATENT ABSTRACTS 651

between fibers exceeding about 20 mum in dia- meter, and has a Frazier Air Permeability in CFM/ft2 at 0.5" H20 of about 215 for a cloth of thickness of about 40 mils. The cloth has a large surface area for binding to an antibody or antigen and can accommodate a large volume of liquid per surface area and has minimum flow resistance. The cloth containing an immobilized antibody or antigen may be used to carry out an enzyme immunoassay by contacting the cloth with a sample containing an antigen or antibody, incubating the cloth with an enzyme-antibody conjugate and then reacting enzyme boundto the cloth with a chromogenic substrate-indicator to produce a visible color. Other immunoassay em- bodiments may also be carried out and a control cloth can be used such that a difference in color from that obtained with the control cloth deter- mines the amount of antigen or antibody present in a sample.

5122602

C H R O M O G E N I C M E R O C Y A N I N E E N Z Y M E S U B S T R A T E S

Paul F Corey, M Teresa Yip assigned to Miles Inc

Chromogenic merocyanine enzyme substrate compounds of the general formula: See Patent for Chemical Structure where Y is an enzymatically-cleavable group such as a radical of a sugar, carhoxylic acid, amino acid, peptide, phosphoric acid, or sulfuric acid; A and B represent residues that complete 5- or 6- membered ring systems; R1 is substituted or unsubstituted alkyl; R2 and R3, independently, are hydrogen or lower alkyl; m, n, and p, which can be different, are integers from 0 through 3 provided that m + n + p must be at least 2; and X is an appropriate counterion (anion).

5122462

P R O C E S S F O R T H E E N Z Y M A T I C P R E P A R A T I O N O F O P T I C A L L Y -

A C T I V E C Y A N O H Y D R I N S

Peter Miethe, Maria-Regin Kula, lngeborg M Stuertz, Christia Wandrey, Udo Kragl, Halle, Federal Republic Of Germany assigned to For- schungszentrum Juelich GmbH

The preparation of optically-active cyano- hydrins from the corresponding oxo compounds is disclosed. The reaction of the oxo compounds with hydrocyanic acid is carried out in an organic solvent in the presence of (R)- or (S)- oxynitrilase (4.1.2.10) and (4.1.2.11), respectively, being solubilized in a lyotropic liquid crystal. Compounds which upon hydrolysis produce increased pH values are excluded as surface active agents. Preferably, surface active a- gents, the organic solvent and an aqueous buffer solution with a pH of 3 to 6, are mixed together to obtain a liquid crystal/organic solvent two- phase system. The liquid crystal is preferably fixed on a porous support, in particular a glass support. The reaction is carried out in a flow- through reactor which contains the liquid crystal in the abovementioned form or in thin layers ad- jacent to narrow flow channels, the borders of which are liquid permeable and through which the substrate-containing solvent is passed.

5130237

S U B S T R A T E C O N V E R S I O N W I T H AN E N Z Y M E I M M O B I L I Z E D O N

AN U L T R A F I L T R A T I O N M E M B R A N E

Ronald L Thomas, Daniel L McKamy assigned to Clemson University

A process is disclosed for chemically converting a substrate into its reaction products and im- mediately thereafter physically separating the reaction products in a continuous operation. The process is carried out with a bioreactor having an ultrafiltration membrane containing an immobilized chemical agent which is preferably an enzyme. The bioreactor is prepared by securing an ultrafiltration membrane to an in- side wall of a porous tubular support and chemically bonding an enzyme to an inner surface of the membrane. The enzyme is preferably bonded to the membrane by chelation and the membrane may be a polymeric membrane or a metal oxide membrane. To convert a substrate, a substrate- containing feed stream is preferably flowed tangentially along the inner surface of the membrane containing the immobilized enzyme. Suf- ficiently small reaction products filter through pores of the membrane and larger reaction products are retained by the membrane. In a preferred embodiment, contacting of fresh fruit juice with pectinase immobilized on the membrane results in pectinase treatment or the juice and immediate extraction and clarification of the


juice. In another embodiment, the enzyme immobilized is glucoamylase and corn dextrins are converted to reducing sugars.

5132219

E N Z Y M E S F R O M R H O D O C O C C U S

R H O D O C H R O U S S T R A I N A T C C N O . 53968 , B A C I L L U S

S P H A E R I C U S S T R A I N A T C C N O . 53969 A N D M I X T U R E S T H E R E O F

F O R C L E A V A G E O F O R G A N I C C - S B O N D S O F C A R B O N A C E O U S

M A T E R I A L

5135869

S E L E C T I V E E N Z Y M A T I C D E G R A D A T I O N O F B E T A -

L A C T O G L O B U L I N C O N T A I N E D IN C O W ' S M I L K - S E R U M

P R O T E I N

Tetsuo Kaneko, Tadashi Kojima, Tamotsu Kuwata, Yoshiro Yamamoto, Saitama, Japan assigned to Meiji Milk Products Company Limited

This invention relates to a method of selectively degrading beta-lactoglobulin contained in cow's milk-serum protein by using a specific enzyme capable of selectively degrading beta- lactoglobulin.

John J Kilbane assigned to Institute of Gas Technology

An extract of membrane fragments, an enzyme, and a composition of enzymes associated with cell membranes of Rhodococcus rhodochrous strain ATCC No. 53968 and Bacillus sphaericus strain ATCC No. 53969 is prepared which have the ability to selectively react with organic sulfur of sulfur-containing organic carbonaceous material by cleavage or organic C-S bonds.

5137818

I M M O B I L I Z E D B I O C A T A L Y S T S

5134072

Abraha Harder, Ben R DeHaan, Plaat Johannes B Van Der, Marsha Cummings, Berkel en Rodenrijs, Netherlands assigned to Gist- Brocades N V

P R E P A R A T I O N O F A W A T E R - S O L U B L E M O D I F I E D E N Z Y M E

BY C O V A L E N T L Y B O N D I N G A N E N Z Y M E T O A P O L Y U R E T H A N E

P R E - P O L Y M E R ] B I S U L F I T E A D D U C T

Ulrich Eicken, Wilhelm Tischer, Korschen- broich, Federal Republic Of Germany assigned to Boehringer Mannheim GmbH

A water-soluble polyurethane-modified enzyme is prepared by reaction of an enzyme in aqueous solution with an aqueous solution of a water- soluble polyurethane pre-polymer/bisulfite adduct at a pH greater than 7 to covalently bond the enzyme to the adduct. Preferably, a ratio of enzyme to pre-polymer/bisulfite adduct of from 1:10 to 10:1 is used.

Immobilized water-insoluble biocatalysts in particulate form comprise living cells, particularly yeast, dispersed in a cross-linked gelling agent. An enzyme, particularly amyloglucosidase, may be co-immobilized in the particles. These particles are prepared by suspen- ding the living cells in an aqueous solution of a gelling agent, dispersing this suspension in a water immiscible organic liquid to form a suspension in the liquid of aqueous particles comprising the living cells and gelling agent, gelling the gel and cross-linking the gelling agent. It is found that when living cells such as microbial cells and especially yeast are immobilized in this way, that surprisingly, not only is their viability retained, but the ability of yeast cells to produce ethanol under continuous fermentation condi- tions is significantly improved. Specific strains of Saccharomyces cerevisiae, suitable for ira-

• mobilization in this way, are described.

PATENT ABSTRACTS 653

5139942

M E T H O D F O R P R O D U C I N G T H E N D E I R E S T R I C T I O N

E N D O N U C L E A S E A N D M E T H Y L A S E

Jack S Benner assigned to New England Biolabs Inc

The present invention is directed to a method for cloning and producing the Nde I restriction endonuclease by 1) introducing the restriction endonuclease gene from Neisseria denitrificans into a host whereby the restriction gene is ex- pressed; 2) fermenting the host which contains the vector encoding and expressing the Nde I restriction endonuclease, and 3) purifying the Nde I restriction endonuclease from the fermen- ted host which contains the vector encoding and expressing the Nde I restriction endonuclease activity.

Disclosed are methods and compositions for the identification, characterization and inhibition of farnesyl protein transferases, enzymes involved in the farnesylation of various cellular proteins, including cancer related ras proteins such as p21 ras. One farnesyl protein transferase which is disclosed herein exhibits a molecular weight of between about 70,000 and about 100,000 upon gel exclusion chromatography. The enzyme appears to comprise one or two subunits of ap- proximately 50 kDa each. Methods are disclosed for assay and purification of the enzyme, as well as procedures for using the purified enzyme in screening protocols for the identification of possible anticancer agents which inhibit the enzyme and thereby prevent expression of proteins such as p21ras. Also disclosed is a families of compounds which act either as false substrates for the enzyme or as pure inhibitors and can therefore be employed for inhibition of the enzyme. The most potent inhibitors are ones in which phenylalanine occurs at the third position of a tetrapeptide whose amino terminus is cysteine.

5139944

C O L L A G E N - S P E C I F I C E N Z Y M E W I T H P L A T E L E T A G G R E G A T I O N

I N H I B I T I O N P R O P E R T I E S

Roy Sawyer, Meir Rigbi, Haim Levy, Fuad Iraqi, Swansea, United Kingdom assigned to Biophram (UK) Limited; Yissum Research Development Company of the Hebrew Univer- sity of Jerusal

The collagenase is like tissue collagenases in that it cleaves collagen at a single site. The collagenase which has a molecular weight of about 50 k daltons, specifically inhibits collagen- induced platelet aggregation, with substantially no effect on platelet count or size, and may be used alone or together with another platelet aggregation inhibitor. The latter may also be a leech-derived biochemical, such as an apyrase, or an inhibitor of the release of platelet aggregation factor by leucocytes.

5141851

I S O L A T E D F A R N E S Y L P R O T E I N T R A N S F E R A S E E N Z Y M E

Michael Brown, Joseph L Goldstein, Yuval Reiss assigned to Board of Regents The Univer- sity of Texas System

5143828

M E T H O D F O R S Y N T H E S I Z I N G A N E N Z Y M E - C A T A L Y Z E D

P O L Y M E R I Z E D M O N O L A Y E R

Joseph A Akkara, David L Kaplan, Lynne A Samuelson, Braja K Mandal, Sukant K Tripathy, Ferdinando F Bruno, Kenneth Marx assigned to The United States of America as represented by the Secretary of the Army; University of Massachusetts Lowe

A method for synthesizing enzyme-catalyzed polymers using the Langmuir-Blodgett techni- que. In one embodiment, the process comprises spreading one or more enzyme-polymerizable monomers on a water-miscible solvent. The monomers are sufficiently surface active that they align themselves on the air-solvent interface. Next, pressure is applied to the interface to form a monolayer made up of the monomers. An enzyme is then introduced into the solvent, causing polymerization of the monomers in the monolayer. The polymeric monolayers pro- duced by the present method are easier to process and have reduced cross-linking and branching as compared to similar polymers pro- duced in bulk by enzyme-catalyzed reactions.


5143837

E N Z Y M E C O M P L E X H A V I N G C O L L A G E N O L Y T I C A C T I V I T Y

I S O L A T E D F R O M C R A B S

Vyacheslav Sova, Alexander Strongyn, Olga Klimova, Vadim Stadnikov, Petropavlovsk, Union Of Soviet Socialist Republics assigned to Seatec

An enzyme complex having collenagenolytic activity, comprising a mixture of collegenolytic proteases from crabs is disclosed. The proteases of the complex have the ability to cleave bovine lens capsule collagen type IV, have molecular weights from about 23,000 to 36,000 daltons, and possess a collagenolytic activity of more than about 3.300 Mandl units/nag at a pH of 7.5. The complex is isolated in aqueous solutions without the use of organic solvents from the hepatopancreas of crabs. The crabs from which the complex can be isolated are of the para- lithodes and chinocetes species of crabs.

5143840

S U B S T A N T I A L L Y P U R I F I E D P R O T E O L Y T I C E N Z Y M E F R O M

B A C I L L U S S P E C . U S N I 4828 M E T H O D O F M A K I N G A N D

U S I N G IT

Hansjoerg Rettenmaier, Andreas Kreimeyer, Johannes Perner, Paul Diessel, Gruenstadt,

Federal Republic Of Germany assigned to BASF Aktiengesellschaft

A substantially purified proteolytic enzyme isolated from a culture of Bacillus spec. DSM 4828 which has a molecular weight of 27,000 daltons, an optimum pH of 12, an optimal tempera- ture of 60 degrees C. and which can be used in a detergent composition.

5143847

ENZYME-FIXED BIOREACTOR

Mitsuo Kawase, Yasuko Yoshida, Hitosh Yonekawa, Chita, Japan assigned to NGK In- sulators Ltd

An enzyme-fixed bioreactor, including a reaction column, enzyme-fixed catalyst particles uniformly and densely filled in the reaction column, the catalyst particles being composed of carrier sepiolite particles consisting essentially of sepiolite and an enzyme carried on the surface of the carrier sepiolite particles. The bioreactor has stable heat resistant and chemical properties, high productivity, and is economically superior to prior art, without fear of destruction of the catalyst, short path, and clogging in the reaction column.

enzymes and enzyme systems

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