Novo Nordisk A/s

Helle Fabricius Woldike

Sven Hastrup

Tove Christensen

Carsten Mailand Hjort

Esper Boel

Fred Hagen

Ida Birgitte Huge-Jensen

Thomas Borglum Kjeldsen

Per Franklin Nielsen

Lars Thim

A process for expression of a protein product in Aspergillus oryzae is disclosed. The process comprises transforming Aspergillus oryzae with a vector system comprising DNA sequences encoding functions facilitating gene expression, a suitable marker for selection of transformants, and a DNA sequence encoding the desired protein product. The process enables industrial production of many different polypeptides and proteins in A. oryzae. Examples of such products are chymosin or prochymosin and other rennets, proteases, lipases and amylases. Also disclosed is an effective promoter for expression of a protein in Aspergillus. A preferred promoter is the TAKA-amylase promoter or functional parts thereof. There is also provided a process for the production of a recombinant Humicola lipase. The recombinant Humicola lipase from A. oryzae differs from the native lipase in having a greater glycosylation and in exhibiting an improved thermostability.

Methods for producing Humicola lipases in aspergillus

Trypsin (trypsinogen) may be produced in a filamentous fungus by transforming a filamentous fungus with a vector comprising a DNA sequence encoding protrypsin or a derivative thereof N-terminally fused to a DNA sequence encoding a signal peptide, culturing the transformed filamentous fungus in a suitable culture medium to produce trypsinogen and recovering trypsinogen and/or trypsin from the medium.

Process for producing trypsin (trypsinogen)

A process for expression of a protein product in Aspergillus oryzae is disclosed. The process comprises transforming Aspergillus oryzae with a vector system comprising DNA-sequences encoding functions facilitating gene expression, a suitable marker for selection of transformants, and a DNA-sequence encoding the desired protein product. The process enables industrial production of many different polypeptides and proteins in A. oryzae. Examples of such products are chymosin or prochymosin and other rennets, proteases, lipases and amylases. Also disclosed is an effective promoter for expression of a protein in Aspergillus. A preferred promoter is the TAKA-amylase promoter or functional parts thereof. There is also provided a process for the production of a recombinant Humicola lipase. The recombinant Humicola lipase from A. oryzae differs from the native lipase in having a greater glycosylation and in exhibiting an improved thermostability.

Nucleic acid encoding a recombinant humicola sp. lipase

Humicola lipase produced in aspergillus

The present invention relates to isolated and purified cellulose- or hemicellulose-degrading enzymes, comprising a catalytically active domain, a carbohydrate binding domain and a linking B region which operably links the catalytically active domain and the carbohydrate binding domain, wherein the carbohydrate binding domain has the following core sequence: ##STR1## wherein the enzyme is endogenous to a strain of a fungus other than Trichoderma or Phanerochaete.

Enzyme capable of degrading cellulose or hemicellulose

The present invention relates to cellulose- or hemicellulose-degrading enzymes which is derivable from a fungus other than Trichoderma or Phanerochaete, and which comprises a carbohydrate binding domain homologous to a terminal A region of Trichoderma reesei cellulases, which carbohydrate binding domain comprises the following amino acid sequence. ##STR1## or a subsequence thereof capable of effecting binding of the enzyme to an insoluble cellulosic or hemicellulosic substrate.

A cellulose- or hemicellulose-degrading enzyme which is derivable from a fungus other than Trichoderma or Phanerochaete, and which comprises a carbohydrate binding domain homologous to a terminal A region of Trichoderma reesei cellulases, which carbohydrate binding domain amino acid sequence (.alpha.) or a subsequence thereof capable of effecting binding of the enzyme to an insoluble cellulosic or hemicellulosic substrate.

Enzyme capable of degrading cellullose or hemicellulose

A process for expression of a protein product in Aspergillus is disclosed. The process comprises transforming an Aspergillus strain with a vector system comprising DNA-sequences encoding a promoter including upstream activating sequences derived from an A. niger amylase, a suitable marker for selection of transformants, and a DNA-sequence encoding the desired protein product. The process enables industrial production of many different polypeptides and proteins in Aspergillus, preferably A. niger. Examples of such products are chymosin or prochymosin and other rennets, proteases, lipases and amylases. Also disclosed is an effective promoter for expression of a protein in Aspergillus, preferably Aspergillus niger being derived from a gene encoding an A. niger amylase. The A. niger amylases are the neutral and acid stable .alpha.-amylases and a new amylase not so far described and designated XA amylase. Also disclosed is the novel amylase from A. niger XA amylase.

Growing community of inventors

Lynge, Denmark

Helle Fabricius Woldike