Danisco US Inc.

Genencor International, Inc.

Genencor, Inc.

Other

Eugenio Ferrari

Anita Van Kimmenade

Carol Marie Fioresi

Walter Weyler

M Harunur Rashid

Carole Harbison

David Aaron Estell

Caroline M Peres

Fernando Valle

Cristina Bongiorni

Dennis J Henner

James A Wells

Scott Douglas Power

Marguerite A Cervin

Mark L Stahl

Sierd Bron

Harunur Rashid

The present invention provides cells that have been genetically manipulated to have an altered capacity to produce expressed proteins. In particular, the present invention relates to Gram-positive microorganisms, such asspecies having enhanced expression of a protein of interest, wherein one or more chromosomal genes have been inactivated, and preferably wherein one or more chromosomal genes have been deleted from thechromosome. In some further embodiments, one or more indigenous chromosomal regions have been deleted from a corresponding wild-typehost chromosome.

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The present invention provides methods and compositions for the production of mature proteases in bacterial host cells. The compositions include modified polynucleotides that encode modified proteases, which have at least one mutation in the pro region; the modified serine proteases encoded by the modified polynucleotides; expression cassettes, DNA constructs, and vectors comprising the modified polynucleotides that encode the modified proteases; and the bacterial host cells transformed with the vectors of the invention. The methods include methods for enhancing the production of mature proteases in bacterial host cells e.g.sp. host cells. The produced proteases find use in the industrial production of enzymes, suitable for use in various industries, including but not limited to the cleaning, animal feed and textile processing industry.

Proteases with modified pro regions

Certain aspects of this disclosure relate to a method of amplifying a genomic locus. In certain embodiments, the method may comprise: a) contacting a population of bacterial host cells with an inhibitor of an essential enzyme, where the bacterial host cells comprise a genomic locus of the structure: A-P-M-A, where Aand Aare direct repeats, P comprises a coding sequence for a polypeptide, and M comprises a coding sequence for the essential enzyme; and b) selecting for cells that are resistant to the inhibitor; wherein cells that are resistant to the inhibitor have multiple copies of the amplification unit.

Method for amplifying locus in bacterial cell

This invention relates to modified polynucleotides encoding modified proteases, and methods for altering the production of proteases in microorganisms. In particular, the present invention relates to methods for altering the expression of proteases in microorganisms, such asspecies. The invention discloses modified polynucleotides, vectors, modified polypeptides, and processes for enhancing the production of proteases.

Modified proteases

Modified protease propeptides

The present invention provides methods of altering the production of desired polypeptides in a host cell. In particular, the present invention provides polynucleotides encoding truncated SecG proteins capable of facilitating the secretion of desired proteases by a bacterial host cell, such asspecies, as well as expression vectors and a host cell containing the polynucleotides.

Modified secretion system to increase expression of polypeptides in bacteria

The present invention relates to cells that have been genetically manipulated to have an altered capacity to express and/or produce proteins of interest. In particular, the present invention relates to modified host cells of Gram-positive microorganisms, such asspecies that are capable of overexpressing ymaH. The invention encompasses polynucleotide constructs and expression vectors containing polynucleotide sequences that encode YmaH, and the modified host cells comprising them. In particular, the present invention relates to compositions and methods of overexpressing YmaH for enhancing the expression and production of proteins of interest (e.g., proteases) inspecies.

The present invention provides host cells that have been genetically manipulated to have an enhanced capacity to produce proteins of interest. In particular, the invention relates to modifiedsp. Host cells that have at least one inactivated phr gene. The enhanced production of proteins of interest by the modifiedsp. Host cells is further increased in modifiedsp. Host cells that overexpress YmaH. Methods for producing proteins of interest in the modified host cells are also provided.

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San Bruno, CA, United States of America

Eugenio Ferrari