Linoleic acid biosynthesis and characterization of the. Delta. sup 12 desaturase in insects
Thesis/Dissertation
·
OSTI ID:5160771
De novo biosynthesis of linoleic acid was demonstrated in vivo in 8 of 32 insect species examined, including both holometabolous and hemimetabolous species. The incorporation of (1-{sup 14}C) acetate into linoleic acid was demonstrated by radio-gas-liquid chromatography (radio-GLC), and in selected species by radio-high-performance liquid chromatography, silver nitrate thin-layer chromatography, radio-GLC and GLC linked to mass spectrometry of ozonolysis products. Analysis of the ozonolysis products clearly demonstrated that the entire molecule was labeled and that synthesis of linoleate was de novo from acetate. The in vivo incorporation of (1-{sup 14}C)acetate into lipid was monitored during the final three stadia of both male and female house crickets, Acheta domesticus. Characterization of the {Delta}{sup 12}-desaturase showed that, in the house cricket, this enzyme is microsomal and requires a reduced pyridine dinucleotide as a cofactor, with NADPH the preferred electron donor. The optimal substrate concentration for desaturation is about 40 uM. Addition of the microsomal supernatant, MgCl{sub 2} or ATP did not enhance activity. The form of the substrate for the desaturase, oleic acid, was determined and appears to be a CoA derivative, as is true for most animal desaturases, rather than a complex lipid, as it is in plants.
- Research Organization:
- Nevada Univ., Reno, NV (USA)
- OSTI ID:
- 5160771
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ACETATES
ANIMALS
ARTHROPODS
BIOSYNTHESIS
CARBON 14 COMPOUNDS
CARBOXYLIC ACID SALTS
CARBOXYLIC ACIDS
CHEMICAL COMPOSITION
CHROMATOGRAPHY
COENZYMES
ENZYMES
GAS CHROMATOGRAPHY
INSECTS
INVERTEBRATES
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LINOLEIC ACID
LIQUID COLUMN CHROMATOGRAPHY
MASS SPECTROSCOPY
MONOCARBOXYLIC ACIDS
NADP
NUCLEOTIDES
OLEIC ACID
ORGANIC ACIDS
ORGANIC COMPOUNDS
OXIDOREDUCTASES
OXYGENASES
SEPARATION PROCESSES
SPECTROSCOPY
SUBSTRATES
SYNTHESIS
THIN-LAYER CHROMATOGRAPHY
TRACER TECHNIQUES
59 BASIC BIOLOGICAL SCIENCES
ACETATES
ANIMALS
ARTHROPODS
BIOSYNTHESIS
CARBON 14 COMPOUNDS
CARBOXYLIC ACID SALTS
CARBOXYLIC ACIDS
CHEMICAL COMPOSITION
CHROMATOGRAPHY
COENZYMES
ENZYMES
GAS CHROMATOGRAPHY
INSECTS
INVERTEBRATES
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LINOLEIC ACID
LIQUID COLUMN CHROMATOGRAPHY
MASS SPECTROSCOPY
MONOCARBOXYLIC ACIDS
NADP
NUCLEOTIDES
OLEIC ACID
ORGANIC ACIDS
ORGANIC COMPOUNDS
OXIDOREDUCTASES
OXYGENASES
SEPARATION PROCESSES
SPECTROSCOPY
SUBSTRATES
SYNTHESIS
THIN-LAYER CHROMATOGRAPHY
TRACER TECHNIQUES