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Fatty acid and derivatives production

A fatty acid and derivative technology, applied in biosynthesis, DNA/RNA fragments, recombinant DNA technology, etc., can solve problems such as difficult control of composition

Inactive Publication Date: 2016-05-25
EVONIK DEGUSSA GMBH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Another problem associated with the biotechnological route is the fact that mixtures of products are obtained and thus the composition is difficult to control

Method used

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  • Fatty acid and derivatives production
  • Fatty acid and derivatives production
  • Fatty acid and derivatives production

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Construction of plasmids for the production of fatty acids using R. eutropha

[0059] To construct a plasmid for the production of fatty acids from R. eutropha, 3 synthetic expression cassettes consisting of the following components were synthesized:

[0060] R. eutropha groEL Ribosome binding site of gene (SEQ ID NO: 1), gene encoding thioesterase from Calyx chinensis ChFATB2 (SEQ ID NO:2), which has been 5'-truncated at the plastid targeting sequence, Escherichia coli rrnB The terminator (SEQ ID NO:5) of gene, wherein in the translation of Laueria eutropha, is used for ChFATB2The coding region of is codon-optimized (RBSRegroEL-ChFATB2-T; SEQ ID NO: 6).

[0061] R. eutropha groEL The ribosome binding site (SEQ ID NO: 1) of gene, coding comes from coconut ( Cocos nucifera ) thioesterase gene CnFATB3 (SEQ ID NO:3), Escherichia coli rrnB The terminator (SEQ ID NO:5) of gene, wherein in the translation of Laueria eutropha, is used for CnFATB3 The coding ...

Embodiment 2

[0065] Introduction of plasmids for fatty acid production in Laueria eutropha

[0066] The plasmids from above were transfected into competent E. coli S17-1 cells in which conjugative transfer of plasmids from R. eutropha among other strains was possible. For this purpose, Spotmating conjugation using the respective plasmids was performed (e.g. FRIEDRICH et al., 1981), in which E. coli strain S17-1 was the donor and R. eutropha H16 (reclassified as Alcaligenes eutropha , DSMZ428) and R. eutropha PHB-4 (reclassified as Alcaligenes eutropha, DSMZ541) are recipients. Zygotes were obtained in all cases carrying the respective plasmids and the corresponding strains were named as follows:

[0067] R. eutropha H16PbBr-ChFATB2, R. eutropha H16PbBr-CnFATB3, R. eutropha H16PbBr-UcFATB1, R. eutropha PHB-4-PbBrChFATB2, R. eutropha PHB- 4-PbBrCnFATB3, and R. eutropha PHB-4-PbBrUcFATB1.

Embodiment 3

[0069] Quantification of fatty acids

[0070] Quantification of caprylic acid, 3-hydroxycapric acid, capric acid, lauric acid, 3-hydroxymyristic acid, myristic acid, palmitoleic acid, palmitic acid, oleic acid, and stearic acid in fermentation samples by An internal calibration of HPLC-ESI / MS was performed and internal standards were used: caprylic acid, 3-hydroxycapric acid, capric acid, lauric acid, 3-hydroxymyristic acid, myristic acid, palmitoleic acid, stearic acid D3 Lauric Acid (Methyl-D3, 99%), and D3 of Palmitic, Oleic, Stearic Acids (D3-Methyl, 98%).

[0071] Use the following equipment:

[0072] ●HPLC system: Surveyor (ThermoFisherScientific, Waltham, Massachusetts, USA), composed of SurveyorMS pump, SurveyorAutosamplerPlus and SurveyorSurveyorPDA

[0073] ●Mass spectrometer: TSQVantageHESIII-source (ThermoFisher Scientific, Waltham, Massachusetts, USA)

[0074] HPLC column: XBridge BEHC8, 100x2.1 mm, particle size: 2.5 microns, pore size 130 Å (Waters, Milford...

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Abstract

The present invention relates to at least one method of producing at least one fatty acid and / or derivative thereof from a gas comprising H 2 , CO 2 and O 2 the method comprising the steps of: (a) providing a genetically modified hydrogen oxidising bacterium in an aqueous medium; and (b) contacting the aqueous medium with a gas comprising H2, CO2 and O2 in a weight ratio of 20 to 70: 10 to 45: 5 to 35; wherein the fatty acid comprises at least 5 carbon atoms and wherein the hydrogen oxidising bacterium is genetically modified relative to the wild type bacterium to increase the expression of enzyme E 1 that is capable of catalysing the conversion of acetyl CoA to acyl ACP via malonyl coA and to increase the expression of enzyme E 2 that is capable of catalysing the conversion of Acyl ACP to the fatty acid.

Description

field of invention [0001] The present invention relates to the production of fatty acids by biotechnological means. In particular, the invention relates to the biotechnological production of fatty acids with more than 5 carbon atoms from renewable resources. Background technique [0002] Fatty acids and / or triglycerides have various uses in various industries. In the food industry, for example, fatty acids and / or fat triesters can be used in animal feed and for nutritional supplementation. Fatty acids and / or triglycerides can also be used in the cosmetic and pharmaceutical fields. These applications may require free fatty acids or triglycerides. Natural sources of these fatty acids and / or triglycerides are rather limited. [0003] For example, various polyunsaturated fatty acids (PUFAs) and PUFA-containing triglycerides are mainly obtained from microorganisms such as Mortierella ( Mortierella ) and Schizochytrium ( Schizochytrium ), or oleaginous plants such as soybea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/40C12P7/64
CPCY02P20/59C12P7/6409C12N15/52C12P7/64
Inventor T.哈斯M.珀特S.沙费尔
Owner EVONIK DEGUSSA GMBH
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