Omnigene, Inc.

Roche Vitamins Inc.

Biotechnica International, Inc.

Hoffmann-La Roche Inc.

Dsm Nutritional Products Inc.

Alan Sloma

Janice G Pero

Gerald A Rufo, Jr

John B Perkins

Cathy F Rudolph

Thomas John Erdenberger

Theron Hermann

Randolph T Hatch

Barbara J Sullivan

Nancy M Hannett

Rosalind C Lee

M A Stephens

Vectors and recombinant bacteria for overproducing riboflavin, in which nucleic acid overproducing riboflavin biosynthetic proteins is introduced in the chromosome of the host organism, e.g. at multiple sites and in multiple copies per site. A rib operon having at least five genes is used to make such recombinant bacteria.

Bacterial strains which overproduce riboflavin and methods of use thereof

Nutrient medium for bacterial strains which overproduce riboflavin

Bacterial strains which overproduce riboflavin

A Bacillus cell contains a mutation in the epr gene resulting in inhibition of the production by the cell of the proteolytically active epr gene product; the cell may further contain mutations in the genes encoding proteolytically active residual protease I (RP-I) and proteolytically active residual protease II (RP-II) (mpr).

Protease deletion

A Bacillus cell contains a mutation in the epr gene resulting in inhibition of the production by the cell of the proteolytically active epr gene product; the cell may further contain mutations in the genes encoding proteolytically active residual protease I (RP-I) and proteolytically active residual protease II (RP-II).

A Bacillus cell contains a mutation in the epr gene resulting in inhibition of the production by the cell of the proteolytically active epr gene product; the cell may further contain mutations in the genes encoding proteolytically active residual protease I (bpr) and proteolytically active residual protease II (RP-II).

A Bacillus cell containing a mutation in the residual protease III (rp-III) gene resulting in the inhibition of the production by the cell of proteolytically active RP-III.

Residual protease-III

A vector for high-level expression of a heterologous gene in Bacillus, including a DNA sequence which encodes all or part of the structural gene and contains the promoter and ribosome binding site of a Bacillus coagulans amylase gene; within or immediately downstream of the structural gene is a site for insertion of heterologous DNA; when inserted, the heterologous DNA is in the same translational reading frame as the amylase gene; the heterologous DNA thus is under control of the B. coagulans amylase gene regulatory elements and expressed at high levels. A signal-encoding sequence may be inserted independent of, or along with, the heterologous DNA leading to authentic secretion of the heterologous protein.

Expression of heterologous DNA using the bacillus coagulans amylase gene

A bacterial cell transformed with a gene encoding a desired product and with an enhancing DNA sequence capable of enhancing the production of the desired product in the bacterial cell, the enhancing DNA sequence being further characterized in that it is capable of enhancing the production of an endogenous protease in a Gram-positive bacterial cell.

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Watertown, MA, United States of America

Alan Sloma