Agricultural & Food Research Council

Institute of Food Research

Plant Bioscience Limited

Biotechnology & Biological Sciences Research Council

Michael John Gasson

Arjan Narbad

Michael John Charles Rhodes

Helen Mair Dodd

Nicholas John Walton

A method of producing vanillin comprising the steps of: (1) providing trans-ferulic acid or a salt thereof; and (2) providing trans-ferulate: CoASH ligase activity (enzyme activity I), trans-feruloyl ScoA hydratase activity (enzyme activity II), and 4-hydroxy-3-methoxyphenyl-&bgr;-hydroxy-propionyl SCoA (HMPHP SCoA) cleavage activity (enzyme activity III). Conveniently the enzymes are provided by Fe3 or a mutant or derivative thereof. Polypeptides with enzymes activities II and III and polynucleotides encoding the polypeptides. Use of the polypeptides or the polynucleotides in a method for producing vanillin is also provided.

Production of vanillin

The invention provides a method for making a cell which does not contain a natural nisA gene but expresses a variant nisA gene, said method comprising the steps of providing a cell that contains a natural, chromosomal nisA gene and substituting the natural nisA gene or part thereof with a variant nisA gene at the chromosomal location of the natural nisA gene or part thereof. The invention further provided a process for producing variant nisin and cells for use in the same.

Production of variant nisin

A method of producing vanillin comprising the steps of: (1) providing trans-ferulic acid or salt thereof; and (2) providing trans-ferulate; CoASH ligase activity (enzyme activity I) trans-feruloyl SCoA hydratase activity (enzyme activity II), and 4-hydroxy-3-methoxyphenyl-&bgr;-hydroxy-propionyl SCoA (HMPHP SCoA) cleavage activity (enzyme activity III). Conveniently the enzymes are provided by,Fe3 or a mutant or derivative thereof. Polypeptides with enzymes activities II and III and polynucleotides encoding said polypeptides. Use of said polypeptides or said polynucleotides in a method for producing vanillin.

Nisins

Bacteriophages of food-contaminating or pathogenic bacteria or the lysins thereof are used to kill such bacteria. Examples include lysins from bacteriophages of Listeria monocytogenes and Clostridium tyrobutyricum. Tests for bacterial contamination can be made specific for specific bacteria by using the appropriate bacteriophage or lysin thereof and determining whether cells are lysed thereby.

Viral products

The lysin from a Lactococcus (preferably prolate-headed) bacteriophage is used to lyse bacterial starter cultures during cheese-making. Such bacteriophage include .phi.vML3. In addition, the .phi.vML3 lysin has been characterized and a coding sequence for it has been cloned.

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Norfolk, United Kingdom

Michael John Gasson