4778757 method for the determination of substrates or enzyme activities

100 PATENT ABSTRACTS

organisms in an amount such that when a suitable microorganism is cultivated in such a medium, one or more desaturase enzymes are in- hibited so that higher levels of saturated or less saturated lipids may be produced which may be used in fat compositions as cocoa butter equiva- lents.

4778757

M E T H O D F O R T H E D E T E R M I N A T I O N O F

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

Shinichi Teshima, Noboru Mitsuhida, Yoshitak Nakagiri, Tsuruga, Japan assigned to Toyo Boseki Kabushiki Kaisha

There is provided an improved quantitative analysis for the determination of a substrate or enzyme. In the analysis for the enzyme, it is reac- ted with a known substrate and in the analysis for the substrate, it is reacted with a known en- zyme and then the resultant substance is reacted with an oxidase to produce hydrogen peroxide which is then reacted with a peroxidase, phenol derivatie and coupler to form a colored material which is then subjected to colorimetic analysis. The improvement resides in conducting the fore- going process by adding to the test sample a first reagent containing the oxidase, peroxidase and phenol derivative represented by the formula: See Patent for Chemical Structure (I) wherein R 1 is **a a lower alkyl group having one to five car- bon atoms or **b the group defined in **a above having a hydroxyl or sulfonic acid group, R2 is a hydrogen or halogen atom, a lower alkyl group having one to five carbon atoms, a lower acyl group having one to five carbon atoms, a lower alkylether group having one to five carbon atoms, or a lower alkoxycarbonyl group having one to five carbon atoms, said groups being unsubstituted or substituted with a hydroxyl or sulfonic acid group, and n is 0 to 4. Then, there is added to the above mixture a second reagent containing the coupler and the enzyme.

A preferred method wherein 5-phenyl-4- pentenyl-hydroperoxide (PPHP) is reduced to 5- phenyl-4-pentenyl-alcohol (PPA) by plant and animal peroxidases in the presence of reducing substrates is described. The method also uses related homologs containing 3 to 8 carbon atoms. PHP and PPA are rapidly isolated with solid phase extraction, separated by isocrated reverse phase high performance liquid chromatography, and quantitated with a fixed- wavelength ultraviolet detector. The procedure described in suitable for detecting peroxide reducing enzymes, determining the kinetic pro- perties of heme- and non-heme-containing peroxidases, and evaluating oxidizable com- pounds as reducing substrates for peroxidases. The method identifies compounds which are reducing substrates and also ranks them for rela- tive activity. The method can be used to identify active antithrombotic, antimetastatic, or anti- inflammatory drugs as substrates as well as detect and characterize mammalian peroxidases.

4780410

S A N D W I C H E N Z Y M E I M M U N O A S S A Y F O R P I V K A - I I

W I T H M O N O C L O N A L A N T I - P I V K A - I I A N T I B O D Y

Ichiro Matsuda, Kunihiko Motohara, Kumamoto, Japan assigned to Eisai Co Ltd

A method for use in the assay of PIVKA-II by the enzyme immunoassay using the two antibody-sandwiching method, characterized by using a monoclonal anti-PIVKA-II antibody as solidified antibody for said assay.

4780412

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

E S T A B L I S H E D N O N - C A N C E R O U S C E L L L I N E S

4780281

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

R E D U C I N G S U B S T R A T E A C T I V I T Y

Lawrence J Marnett, Paul E Weller assigned to Wayne State University

Anthony Atkinson, Asgar Electricwala, John B Griffiths, Amy Latter, Patrick A Riley, Peter M Sutton, Salisbury, United Kingdom assigned to Public Health Lab Svc Bd University Coil

Fibrinolytic enzymes are produced and then isolated from non-cancerous established cell lines, designated as BEB or GPK. These fibrinolytic enzymes differ from those produced by malignant cell lines.