Zeagen, Inc.

Synergen Associates, Inc.

Nexstar Pharmaceuticals, Inc.

University Technology Corporation

Universal Foods Corporation

Michael J Yarus

Donald L Heefner

Linda A Burdzinski

Robert J Seely

Craig A Weaver

Sally A Sullivan

Robert V Hageman

Anastasia Khvorova

Randal A Hassler

Richard D Medwid

Ronald J Evans

Beatrice J Kopp

Annette Boyts

Charles H Squires

Dale C Gyure

Hans H Liao

Kathleen S Sniff

Edward W Foster

Mali Illangasekare

The present invention provides nucleic acid molecules capable of binding phospholipid membranes, and more particularly RNA molecules capable of binding and forming channels in biological membranes. In this regard the present invention provides methods for screening nucleic acid molecules that bind phospholipid membranes. The present invention also provides compositions of nucleic acid molecules capable of binding phospholipid membranes, as well as methods employing these compositions to alter the permeability or detectably label phospholipid membranes.

RNA channels in biological membranes

The instant invention provides a method for identifying and preparing nucleic acid ligands capable of facilitating aminoacyl-RNA synthesis. The nucleic acid ligands are identified and produced by exposure of candidate nucleic sequences to a reactant under conditions conducive to reaction, wherein nucleic acid sequences having facilitative activity are able to facilitate or enhance a reaction between the reactant molecule and nucleic acid sequence. Included in the invention are specific RNA ligands capable of facilitating aminoacyl-RNA synthesis identified by the SELEX method.

Method for the production of ligands capable of facilitating

A culture for the production of xanthophylls comprising Nospongiococcum excentricum ATCC 40335 and mutants thereof. The culture having a dry cell weight xanthophyll content of at least about 0.65% and being capable of growing to a cell density of greater than about 40 grams per liter. The culture also comprising: carbon, phosphate, sulfate, iron, magnesium, and nitrogen.

Carotenoid producing culture using Nespongiococcum excentricum

Mutations are induced in a microorganism selected from the species Saccharomyces cerevisiae or from the species Candida flareri. The resulting mutants are cultured in the presence of a fermentation inhibitor, such as acetaldehyde, ephedrine or PAC-dione, to form colonies having resistance to the inhibitor. Cells from the colonies are isolated and tested for yield of phenyl acetyl carbinol (PAC) in a fermentation with benzaldehyde and pyruvate. Yeast cells from the colonies that produce elevated levels of PAC are selected for use in subsequent fermentations. PAC is useful as an intermediate in the preparation of l-ephedrine and d-pseudoephedrine, two well-known medicinal chemicals.

Process for making L-phenyl acetyl carbinol (PAC), microorganisms for

A method of systematically generating and isolating highly flavinogenic strains of Candida famata. The strains Candida famata GA18Y8-6#2 dgr and Candida famata GA18Y8-6#2#11 have been developed and produce yields of ca. 7.0 to 7.5 grams per liter per 6 days. The method includes a combination of iterative mutagenizing steps and protoplast fusion steps performed on the parent strain and the descendant strains which are selected following each step according to a screening protocol.

Efficient riboflavin production with yeast

Strains of yeast of the species Candida famata are disclosed which can produce 10 grams of riboflavin per liter in 6 days, and in particular, strains identified by ATCC Accession Nos. 20849 and 20850. Riboflavin yields of more than 20 grams per liter in 200 hours have been achieved. Strains of the present invention have increased sensitivity to iron inhibition of flavinogenesis and have enhanced riboflavin production per cell at increased iron concentrations in the fermentation medium. The invention also is directed toward a process for selecting improved microorganisms which are resistant to inhibition of growth by depleted medium. Such selected mircoorganisms are then tested for riboflavin overproduction. The present invention is also directed toward a selection process in which mutated microorganisms are cultured in the presence of tubercidin, a purine analog. Mutant strains resistant to tubercidin are then tested for riboflavin over-production. The present invention also includes a process for producing riboflavin by culturing strains of riboflavin overproducing microorganims in a fermentation medium. Increased riboflavin production can be obtained by elevated iron concentrations in the nutrient medium. The fermentation is conducted and after cell growth levels off, riboflavin is recovered from the fermentation medium.

Method for producing riboflavin with Candida famata

Strains of yeast of the species Candida famata are disclosed which can produce 10 grams of riboflavin per liter in 6 days, and in particular, strains identified by ATCC Accession Nos. 20849 and 20850. Riboflavin yields of more than 20 grams per liter in 200 hours have been achieved. Strains of the present invention have increased sensitivity to iron inhibition of flavinogenesis and have enhanced riboflavin production per cell at increased iron concentrations in the fermentation medium.

Riboflavin producing strains of microorganisms

Mutations are induced in a microorganism selected from the species Saccharomyces cerevisiae or from the species Candia flareri. The resulting mutants are cultured in the presence of a fermentation inhibitor, such as acetaldehyde, ephedrine or PAC-dione, to form colonies having resistance to the inhibitor. Cells from the colonies are isolated and tested for yield of phenyl acetyl carbinol (PAC) in a fermentation with benzaldehyde and pyruvate. Yeast cells from the colonies that produce elevated levels of PAC are selected for use in subsequent fermentations. PAC is useful as an intermediate in the preparation of 1-ephedrine and d-pseudoephedrine, two well-known medicinal chemicals.

Process for preparing microorganisms used for making phenyl acetyl

Method for transforming anaerobic microorganisms. Anaerobic microorganisms are induced to form L-forms. Genetic material capable of inducing the desired phenotype is introduced into the L-forms, after which the L-forms may be caused to regenerate their cell wall. These methods are also useful for obtaining desired biological products. Additionally, shuttle vectors capable of transforming both aerobic and anaerobic microorganisms are set forth.

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Boulder, CO, United States of America

Michael J Yarus