Gist-brocades N.v.

Quest International B.v.

Applied Nanosystems B.v.

Unilever Patent Holdings B.v.

Van Den Bergh Foods Co., Division of Conopco, Inc.

Rijksuniverstteit Te Groningen

Gerard Venema

Jan Kok

Adrianus M Ledeboer

Sierd Bron

Girbe Buist

Hilde E Smith

Cornelis Johannes Leenhouts

Jan H Van Ee

Ben P Peeters

Wilhelmus J Quax

Jan Maat

Jan Maarten Van Dijl

Oene Robert Veltman

Lambertus Van Den Burg

Arjan Nauta

Jan W Sanders

Josephus Mauritius Van Der Vossen

The invention relates to surface display of proteins on microorganisms via the targeting and anchoring of heterologous proteins to the outer surface of cells such as yeast, fungi, mammalian, plant cells, and bacteria. The invention provides a proteinaceous substance comprising a reactive group and at least one attaching peptide including a stretch of amino acids having a sequence corresponding to at least a part of the consensus amino acid sequence listed in FIG.and further includes a method for attaching a proteinaceous substance to the cell wall of a microorganism comprising the use of the attaching peptide.

Attaching substances to microorganisms

The invention relates to surface display of proteins on micro-organisms via the targeting and anchoring of heterologous proteins to the outer surface of cells such as yeast, fungi, mammalian and plant cells, and bacteria. The invention provides a proteinaceous substance comprising a reactive group and at least one attaching peptide which comprises a stretch of amino acids having a sequence corresponding to at least a part of the consensus amino acid sequence listed in FIG.and comprises a method for attaching a proteinaceous substance to the cell wall of a micro-organism comprising the use of said attaching peptide.

Attaching substances to micro-organisms

The present invention provides genes encoding variants of metallo-endopeptidases that have been engineered to be resistant to prolonged boiling while having maintained their enzymatic performance at much lower temperatures. In addition, thermal stability of the metallo-endopeptidases is highly dependent on calcium at concentrations in the mM range. The invention further provides active metallo-endopeptidases variants whose stability depending on calcium concentration can be changed so as to provide metallo-endopeptidases that are calcium dependent or independent. The invention also provides genes that encode boiling-resistant metallo-endopetidases whose stability depending on calcium concentration can be changed. The invention also provides vectors and cells comprising these genes and proteases produced through these genes, vectors and/or cells. In particular variants with the above described properties are provided of thermolysin-like proteases such as produced by (TLP-ste) and (thermolysine). Boiling-resistant and calcium independent or dependent metallo-endopeptidases can be applied in several industrial processes, for instance in the preparation of the artificial sweetener aspartame, but also in the leather industry, in breweries and in the production of protein hydrolysates.

Metallo-endopeptidases

Complex inducible promoter system derivable from a phage of a lactic

Salt-inducible promoter derivable from a lactic acid bacterium, and its

Process for the lysis of a culture of lactic acid bacteria by means of a

The claimed invention is drawn to a recombinant plasmid which can replicate in Bacillus subtilis, Escherichia coli, and lactic acid Streptococcus bacteria comprising the replication of origin from Streptococcus cremoris plasmid pWV01 as its origin of replication, in addition to coding marker genes and genes of interest which code for improved fermenting properties.

Plasmids replicatable in Bacillus subtilis, E. coli and lactic acid

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

Gerard Venema