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Dual purposes of linoleate isomerase in aspects of dehydrogenation and isomerism

A linoleic acid isomerase, linoleic acid technology, applied in the directions of isomerase, botanical equipment and methods, biochemical equipment and methods, etc.

Inactive Publication Date: 2011-11-23
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the t10c12-CLA component, there are no reports of heterologous synthesis of functional genes in animal cells or transgenic animals

Method used

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  • Dual purposes of linoleate isomerase in aspects of dehydrogenation and isomerism
  • Dual purposes of linoleate isomerase in aspects of dehydrogenation and isomerism
  • Dual purposes of linoleate isomerase in aspects of dehydrogenation and isomerism

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1. Stable expression of exogenous pai gene in NIH / 3T3 cells

[0035] 1. Construction of eukaryotic gene expression structure

[0036] According to the published original gene sequence of P. acnes linoleate isomerase pai (GenBank accession number AX062088), the original DNA sequence was codon-optimized through genetic engineering methods; at the same time, the start codon ATG was added Known as Kozak sequence oligonucleotides, such as GCCACC, GNN is added after ATG to meet Kozak rules. In this embodiment, a glycine codon GGA is added after the start codon ATG; the coding sequence of histidine tag is added before the stop codon TAA. The optimized nucleotide sequence is the DNA sequence obtained by inserting 6 histidine codons between the 1275 and 1276 positions of sequence 1 in the sequence listing, and adding GCCACC before ATG (the first position of sequence 1). The 5th and 6th positions of sequence 1 are G and A respectively. Among the inserted 6 histidine codons,...

Embodiment 2

[0044] Example 2: Fatty acid determination and CLA biosynthesis guided by exogenous pai gene

[0045] 1 untransfected control cell NIH / 3T3, 1 GPF control cell obtained in Example 1, and 9 positive Pai cell lines obtained in Example 1 at 2×10 per ml. 5 The density of each cell is inoculated into a Φ100-mm culture dish and cultured separately.

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Abstract

The invention discloses dual purposes of linoleate isomerase in aspects of dehydrogenation and isomerism. A bifunctional enzyme PAI is used as delta-12 fattyaciddehydrogenase and linoleate isomerase or delta-12 fattyaciddehydrogenase; and the bifunctional enzyme PAI is protein a) or protein b), wherein the protein a) has amino acid sequences shown as a sequence 2 in a sequence table; and the protein b) has an amino acid residue sequence which is derived from the sequence 2 by substituting and / or losing and / or adding one or more amino acid residues, and has the activity of the delta-12 fattyaciddehydrogenase and linoleate isomerase or the delta-12 fattyaciddehydrogenase. The invention provides a basis and a method for further researching pai gene functions and performing related applicability research on the biosynthesis of t10c12-CLA (conjugated linoleic acid) by using the bifunctional enzyme PAI.

Description

Technical field [0001] The present invention relates to the dual use of linoleic acid isomerase in dehydrogenation and isomerization. Background technique [0002] Conjugated linoleic acid, English name Conjugated linoleic acids, abbreviated as CLA, refers to linoleic acid (C 18 H 32 O 2 ) Is a collective term for isomers composed of a group of positions and geometric isomers. The characteristic of conjugated linoleic acid is that the molecule contains a pair of conjugated carbon-carbon double bonds, one is cis and the other is trans. There are at least 28 isomers of conjugated linoleic acid. Among them about the cis-9, trans-11 (cis-9, trans-11; c9t11-CLA) and cis-10, trans-12 (trans-10, cis-12; t10c12-CLA) isomers There are relatively many studies. c9t11-CLA has anti-cancer properties, can reduce the incidence of cardiovascular diseases, improve immune function, resist diabetes, etc.; t10c12-CLA is mainly involved in controlling the synthesis of body fat and can reduce the in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/90C12N15/61C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C12N7/01C12P7/64
Inventor 苟克勉李世丽
Owner CHINA AGRI UNIV
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