Abstract: A method of increasing recovery of active enzyme produced in a plant is provided where the enzyme requires a transitional metal cofactor for activation. The metal cofactor is supplied to the enzyme during plant development, during extraction, or after extraction. Recovery of active enzyme is also provided by incubating the extracted enzyme at a non-enzyme degrading temperature. Addition of a negative ion salt further improves active enzyme recovery. Optimum salt concentrations for recovery of laccase from plants using copper solutions is provided.

Abstract: Increased expression of heterologous proteins in a plant is achieved by introducing the nucleotide sequence encoding the heterologous protein into a plant that has high oil seed content compared to low oil seed and/or is introduced into a plant having reduced alcohol soluble proteins in the endosperm. The nucleotide sequence may be introduced by direct transformation into the plant, or by direct transformation into another plant, and crossing with the high oil plant or plant have reduced levels of alcohol soluble protein in the endosperm. Further, the nucleotide sequences may be introduced into one or both of the high oil plant and the plant having reduced levels of alcohol soluble proteins, and the plants may be crossed to result in a progeny having even further increased expression levels of the heterologous protein.

Abstract: A method of increasing recovery of active enzyme produced in a plant is provided where the enzyme requires a transitional metal cofactor for activation. The metal cofactor is supplied to the enzyme during plant development, during extraction, or after extraction. Recovery of active enzyme is also provided by incubating the extracted enzyme at a non-enzyme degrading temperature. Addition of a negative ion salt further improves active enzyme recovery. Optimum salt concentrations for recovery of laccase from plants using copper solutions is provided.

Abstract: Production of proteases in plants is set forth, whereby heterologous DNA encoding the protease is introduced into the plant and expression of the protein achieved. By such methods, expression is achieved in plants wherein the plant cell is not damaged, the protein can be recovered without contamination by other proteases, and can be expressed at levels such that commercial production of the enzyme is obtained. Expression levels can be at 0.1% of total soluble protein of the plant, or higher.

Abstract: A method for commercial production of aprotinin entails heterologous expression of the protein in plants, preferably at a level such that aprotinin accounts for at least 0.1% of the total extracted protein. An aprotinin-expressing plant also has potential of increased insecticidal resistance by virtue of producing the protein. A genetic map of the integration locus allows identification of plants derived from the transgenic plant. This approach also reveals genetic loci on a plant chromosome that support high levels of gene expression and can be used as site of integration for expression of other genes of interest.

Abstract: A method for commercial production of GUS entails heterologous expression of the protein in plants, in native conformation, at an expression level such that avidin represents at least 0.1% of total extracted protein. A genetic map of the integration locus allows for the identification of the GUS-expressing plant. Genetic loci on a plant chromosome are revealed that support high levels of GUS expression and that can be used as a site of integration for high level expression of other genes of interest.

Abstract: A method for commercial production of avidin entails heterologous expression of the protein in plants, in native conformation, at an expression level such that avidin represents at least 0.1% of total extracted protein. A genetic map of the integration locus allows for the identification of the avidin-expressing plant. Genetic loci on a plant chromosome are revealed that support high levels of avidin expression and that can be used as a site of integration for high level expression of other genes of interest.