Production and use of inducible enzymes from trichoderma and bacteria for control of plant pests and for industrial processes
First Claim
1. A method of increasing expression of native extracellular inducible microbial enzymes from Trichoderma or Gliocladium species, said method comprising:
- culturing Trichoderma or Gliocladium species in a medium containing material sufficient to induce expression of native extracellular inducible microbial enzymes, low levels of carbohydrates, and/or low levels of reduced nitrogen compounds under conditions effective to increase expression of native fungal extracellular inducible microbial enzymes.
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Abstract
The present invention relates to a method of increasing the expression of native extracellular inducible microbial enzymes from fungal sources using particular culture media and a method of repressing native proteins in fungi and enhancing the production of proteins encoded by transgenes in fungi under the control of an inducible promoter. The present invention also relates to mixtures of extracellular inducible enzymes from a fungal and a bacterial source as well as their use in a method for the degradation of chitinous substrates. Methods of releasing N-acetylglucosamine from chitinous sources and enhancing purity of a heterologous recombinant protein expressed in a culture of fungi are also disclosed.
24 Citations
54 Claims
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1. A method of increasing expression of native extracellular inducible microbial enzymes from Trichoderma or Gliocladium species, said method comprising:
culturing Trichoderma or Gliocladium species in a medium containing material sufficient to induce expression of native extracellular inducible microbial enzymes, low levels of carbohydrates, and/or low levels of reduced nitrogen compounds under conditions effective to increase expression of native fungal extracellular inducible microbial enzymes. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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19. A method, in a culture of fungi, of repressing expression of native proteins and enhancing expression of proteins encoded by transgenes, said method comprising:
culturing fungi, which have been transformed with at least one transgene controlled by a promoter, in a medium containing an inducer of the promoter, high levels of carbohydrates, and/or high levels of reduced nitrogen compounds under conditions effective to repress expression of native proteins and to enhance expression-of proteins encoded by the at least one transgene. - View Dependent Claims (20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30)
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31. A composition comprising:
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a fungal source of a extracellular inducible microbial enzyme and a bacterial source of a extracellular inducible microbial enzyme. - View Dependent Claims (32, 33, 34)
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35. A method of releasing N-acetylglucosamine from a chitinous source comprising:
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treating a chitinous source with a composition comprising;
a fungal source of a extracellular inducible microbial enzyme and a bacterial source of a extracellular inducible microbial enzyme, said treating being carried out under conditions effective to release N-acetylglucosamine from the chitinous source. - View Dependent Claims (36, 37, 38, 39)
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40. A method of repeatedly degrading different chitinous substrates comprising:
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(a) treating a first chitinous substrate with a composition comprising;
a fungal source of a extracellular inducible microbial enzyme and/or a bacterial source of a extracellular inducible microbial enzyme, under conditions effective to produce chitin;
(b) terminating step (a);
(c) treating a second chitinous substrate with the same composition used to carry out step (a) under conditions effective to produce chitin. - View Dependent Claims (41, 42, 43, 44)
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45. A method of increasing expression of native extracellular inducible microbial enzymes from Trichoderma or Gliocladium species, said method comprising:
culturing Trichoderma or Gliocladizim species in a medium containing material sufficient to induce expression of native extracellular inducible microbial enzymes, a carbon source not repressive of expression of fungal extracellular inducible microbial enzymes, and/or low levels of reduced nitrogen compounds under conditions effective to increase expression of native fungal cell extracellular inducible microbial enzymes. - View Dependent Claims (46, 47, 48, 49, 50, 51, 52, 53)
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54. A method of enhancing purity of a heterologous recombinant protein expressed in a culture of fungi, said method comprising:
culturing the recombinant fungi in a medium containing nitrate as the medium'"'"'s sole nitrogen source under conditions effective to induce expression of the heterologous protein while reducing expression of other proteins.
Specification