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Dioxygenases catalyzing asymetric cleavage of beta-carotene

A carotene and dioxygenase technology, applied in the direction of oxidoreductase, enzyme, enzyme, etc., can solve the problem of increased susceptibility

Inactive Publication Date: 2003-06-18
SYNGENTA PARTICIPATIONS AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Additionally, while vitamin A deficiency is not the ultimate determinant of death, it is associated with potentially fatal afflictions such as diarrhea, respiratory disease, and increased susceptibility to childhood illnesses such as measles

Method used

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  • Dioxygenases catalyzing asymetric cleavage of beta-carotene
  • Dioxygenases catalyzing asymetric cleavage of beta-carotene
  • Dioxygenases catalyzing asymetric cleavage of beta-carotene

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Embodiment

[0172] Example plasmid construct Construction of Escherichia coli strain accumulating β-carotene

[0173] A plasmid carrying the β-carotene biosynthesis gene from Erwinia herbicola was constructed using the vector pFDY297. pFDY297 is a derivative of pACYC177 (486-3130 bp) into which bp 1-485 of pBluescriptSK was introduced. To clone the β-carotene biosynthesis gene from Erwinia herbophilus, appropriate endonuclease restriction sites were introduced at both ends of the PCR product. The crtE gene was first inserted into pFDY297. Primers used: 5'-GCGTCGACCGCGGTCTACGGTTAACTG-3' (SEQ ID NO: 3) and 5'-GGGGTACCCTTGAACCCAAAAGGGCGG-3' (SEQ ID NO: 4) and Expand PCR System (Boehringer, Mannheim, Germany), CrtE was amplified by PCR from pBL376 (Hundle BS et al., Mol. Gen. Genet., 245:406-416, 1994), which encodes the entire carotenoid biosynthetic gene cluster from Erwinia herbivora. The PCR product was digested with KpnI and SalI and ligated into the appropri...

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Abstract

The present invention provides means and methods of transforming bacteria, fungi including yeast, animal and plant cells, seeds, tissues and whole plants in order to yield transformants capable of expressing a beta -carotene dioxygenase and accumulating vitamin A aldehyde. The present invention further provides means and methods to biotechnically produce retinoids using cells, tissues, organs or whole organisms which natively or after transformation accumulate beta -carotene or which take up beta -carotene from the medium. The present invention also provides DNA molecules encoding beta -carotene dixoygenases derived from different sources and taxonomic groups of living organisms designed to be suitable for carrying out the method of the invention, and plasmids or vector systems comprising said molecules. Furthermore, the present invention provides transgenic bacteria, fungi including yeast, animal and plant cells, seeds, tissues and whole plants that display an improved nutritional quality or physiological condition and contain such DNA molecules and / or that have been generated by use of the methods of the present invention.

Description

[0001] The present invention relates to the field of transformation of bacterial, yeast, fungal, insect, animal, and plant cells, seeds, tissues, and whole organisms. More specifically, the present invention relates to the integration of recombinant nucleic acid sequences encoding one or more specific enzymes of the carotenoid / retinoid biosynthetic pathway into suitable host cells or organisms, which after transformation exhibit the desired phenotype and can be used, for example, in commercial production. In addition, the present invention provides the realization of oxidative cleavage C by biotechnology 40 Means and methods for producing different characteristic metabolites of the carotenoid / retinoid pathway. Background of the invention [0002] Vitamin A (retinol) and its derivatives (retinal, retinoic acid) (the term "retinoid" is used throughout the specification) represent a group of chemical compounds involved in a broad range of fundamental physiol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H5/00A01K67/027A01K67/033A61K31/7088A61K38/00A61K39/395A61K48/00A61P3/02A61P27/02A61P35/00A61P35/02C07K16/40C12N1/15C12N1/19C12N1/21C12N5/10C12N9/00C12N9/02C12N9/04C12N15/09C12N15/82C12P23/00C12R1/01C12R1/19C12R1/645C12R1/685
CPCC12N15/8243C12P23/00C12N15/825C12N9/0069A61P27/02A61P3/02A61P35/00A61P35/02
Inventor J·冯林提格K·沃格特
Owner SYNGENTA PARTICIPATIONS AG