Danisco US Inc.

Gist-brocades N.v.

Genencor International, Inc.

Danisco US Inc., Genencor Division

Sierd Bron

Wilhelmus J Quax

Eugenio Ferrari

Jan Maarten Van Dijl

Joerg Muller

Jan D H Jongbloed

Gerard Venema

Hilde E Smith

Jan H Van Ee

Ben P Peeters

Harold Tjalsma

Described herein are methods to enhance protein secretion in a host cell. In preferred embodiment, the host cell is a gram-positive microorganism such as a. In another preferred embodiment, the host cell is a gram-negative microorganism. Preferably the gram-negative microorganism is anor a member of the genus. Protein secretion may be enhanced by the overexpression of protein components of the Tat pathway. Alternatively, secretion of foreign proteins can be selectively enhanced by forming a chimeric polypeptide comprising a tat signal sequence and the protein of interest. In a preferred embodiment, the tat signal sequence is selected from phoD or LipA.

[object Object]

The present invention provides methods and compositions for the efficient and enhanced secretion of a protein of interest from a host cell. In specific, proteins are secreted through the Sec-dependent pathway, involving the spoIIIJ and/or yqjG gene product(s). In some embodiments, expression of the spoIIIJ and/or yqjG gene product(s) is modulated by a promoter operably linked to the gene.

Oxa1p enhanced protein secretion

The subject invention describes the cloning and overexpression of leader peptidase genes. A method for isolating a leader peptidase gene is disclosed. Overexpression of the signal peptidase in a suitable host species leads to an enhanced rate of protein processing.

Processing of proteins

Novel vectors are provided for identifying secretory signal sequences from DNA fragments of unicellular microorganisms. The plasmids comprise a multiple cloning site with restriction sites in reading frame with a structural gene which permits rapid screening of the secreted expression product. Optionally, the vectors may include a promoter region upstream from the multiple cloning site. The invention is exemplified with Bacillus. Specific secretory signal sequences have been isolated with those vectors, allowing for efficient secretion into the supernatant, and not just to the periplasmic space to provide proteins in economically high yields. Secretory sequences are provided superior to other previously known sequences.

Secretory signal selection vectors for extracellular protein synthesis

Novel vectors are provided for identifying secretory signal sequences from DNA fragments of unicellular microorganisms. The plasmids comprise a multiple cloning site with restriction sites in reading frame with a structural gene which permits rapid screening of the screted expression product. Optionally, the vectors may include a promoter region upstream from the multiple cloning site. The invention is exemplified with Bacillus. Specific secretory signal sequence have been isolated with those vectors, allowing for efficient secretion into the supernatant, and not just to the periplasmic space to provide proteins in economically high yields. Secretory sequences are provided superior to other previously known sequences.

Growing community of inventors

Haren, Netherlands

Sierd Bron