Bristol-myers Squibb Compnay

Thomas J Franceschini

Suo Win Liu

Shu-Jen Chiang

Michael Politino

Paul M Skonezny

Li You

William V Burnett

Sean M Tonzi

Zizhuo Xing

Gregory L Stein

Nigel Ian Bowers

Ross Binder

Joanne L Brown

Wendy L Anderson

Jonathan D Basch

Bi-cistronic plasmids used for the expression of formate dehydrogenase (FDH) and modified phenylalanine dehydrogenase (PDHmod) are provided.

Genetically stable plasmid expressing PDH and FDH enzymes

The present invention relates to a process for preparing (2R,3S)-1-halo-2-hydroxy-3-(protected)amino-4-substituted butane comprising contacting a mutagenizedhaving ATCC deposit no. PTA-6648 with (3S)-1-halo-2-oxo-3-(protected)amino-4-substituted butane substrate. The present invention also relates to a process comprising mixing (2R,3S)-1-halo-2-hydroxy-3-(protected)amino-4-substituted butane with at least one base in the presence of at least one solvent to produce a reaction mixture containing (2R,3S)-1,2-epoxy-3-(protected)amino-4-substituted butane. The present invention further relates to a process comprising crystallizing the (2R,3S)-1,2-epoxy-3-(protected)amino-4-substituted butane out of the reaction mixture by concurrently adding water and the reaction mixture together. The present invention is also directed to a mutagenizedhaving ATCC deposit no. PTA-6648.

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Process for preparing (2R,3S)-1,2-epoxy-3-(protected)amino-4-substituted butane and intermediates thereof

The invention provides a glutaryl 7-ACA amidase fromstrain BS-203, which catalyzes the hydrolysis of 7-&#x3b2;-(4-carboxybutanamido)-cephalosporanic acid to 7-aminocephalosporanic acid and glutaric acid. The invention also provides nucleic acid sequences, vectors, and host cells useful in the production of this amidase. The glutaryl 7-ACA amidase can be used for the preparation of 7-aminocephalosporanic acid.

Glutaryl cephalosporin amidase from Pseudomonas diminuta BS-203

The invention provides a glutaryl 7-ACA amidase from strain BS-203, which catalyzes the hydrolysis of 7-&bgr;-(4-carboxybutanamido)-cephalosporanic acid to 7-aminocephalosporanic acid and glutaric acid. The invention also provides nucleic acid sequences, vectors, and host cells useful in the production of this amidase. The glutaryl 7-ACA amidase can be used for the preparation of 7-aminocephalosporanic acid.

High expression vectors for expression of heterologous genes in Escherichia coli. The expression vectors contain, the tac promoter, an intergenic region, a restriction site, and, optionally, groES DNA.

High expression Escherichia coli expression vector

A D-amino acid oxidase promoter isolated from T. variabilis has been incorporated into S. cerevisiae expression vectors to increase the expression of heterologous genes. The heterologous promoter has been placed upstream of selective markers, auxotrophic nutritional and/or dominant selectable markers. Because the promoter is from another yeast specie, it is weakly recognized by and S. cerevisiae. To compensate for the weak transcripts of selectable marker genes transcribed from the weak heterologous promoter, the plasmid containing the gene of the interest is amplified leading to a higher expression of protein of interest.

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Cicero, NY, United States of America

Thomas J Franceschini