L-Serine → Arachidonic Acid

HRID 1629664ノート

反応式

HRID 1629664 の反応式
N[C@H:2]([C:7]([OH:9])=[O:8])[CH2:3][CH:4]([CH3:6])C.O=[CH:11][C@@H:12]([C@H:14]([C@@H:16]([C@@H:18]([CH2:20]O)O)O)O)O.S(O)(O)(=O)=O.N1[C:35](N)=[C:34]2[C:30](N=CN2)=NC=1.N[C@H:38]([C:46](O)=O)[CH2:39][CH2:40][CH2:41]NC(=N)N.N[C@H:50](C(O)=O)CC(O)=O.N[C@H](C(O)=O)CCC(O)=O.N[C@H](C(O)=O)CC1N=CNC=1.N[C@H](C(O)=O)CCCCN.N[C@H](C(O)=O)CCSC.N[C@H](C(O)=O)CC1C=CC=CC=1.N[C@H](C(O)=O)CO.N[C@H](C(O)=O)CC1C=CC(O)=CC=1.N[C@H](C(O)=O)C(C)C.N[C@H](C(O)=O)[C@@H](C)O.N[C@H](C(O)=O)CC1C2C(=CC=CC=2)NC=1>>[C:7]([OH:9])(=[O:8])[CH2:2][CH2:3][CH2:4]/[CH:6]=[CH:11]\[CH2:12]/[CH:14]=[CH:16]\[CH2:18]/[CH:20]=[CH:30]\[CH2:34]/[CH:35]=[CH:46]\[CH2:38][CH2:39][CH2:40][CH2:41][CH3:50]

参加物

16

反応物 15

HCID 5951L-SerineHCID 5960L-Aspartic AcidHCID 5962L-LysineHCID 6057L-TyrosineHCID 6106L-LeucineHCID 6137L-MethionineHCID 6140L-PhenylalanineHCID 6287L-ValineHCID 6288L-ThreonineHCID 6305L-TryptophanHCID 33032L-Glutamic AcidHCID 107526(2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanalHCID 118738Adenine SulfateHCID 178418600HCID 178418600、化合物HCID 178419078HCID 178419078、化合物

生成物 1

HCID 444899Arachidonic Acid

実験手順

A DNA fragment of about 0.95 kbp obtained by digesting plasmid pCR-MAGLELO with restriction endonucleases XbaI and SacI was ligated to a DNA fragment of about 7.7 kbp obtained by digesting the vector pESC-LEU (STRATAGENE) with restriction endonucleases XbaI and SacI to generate plasmid pESC-L-GLELO. A DNA fragment of about 1.3 kbp obtained by digesting plasmid pCR-MAΔ5DS with restriction endonuclease XbaI followed by blunt-ending and further digesting it with restriction endonuclease HindIII was ligated to a DNA fragment of about 8.7 kbp obtained by digesting plasmid pESC-L-GLELO with restriction endonuclease ApaI followed by blunt-ending and further digesting it with restriction endonuclease HindIII to generate plasmid pESC-L-GLELO:Δ5. A fragment of about 3.2 kb obtained by digesting this with restriction endonuclease PvuII was inserted into the SmaI site of pUC-LEU2 to generate plasmid pUC-LEU-GLELO:Δ5. S. cerevisiae strain YPH499 (STRATAGENE) was co-transformed with plasmid pUC-URA-Δ12:Δ6 and plasmid pUC-LEU-GLELO:Δ5. Transformed strains were selected by viability on SC-Leu, Ura agar medium (2% agar) containing, per liter, 6.7 g Yeast nitrogen base w/o amino acids (DIFCO), 20 g glucose and 1.3 g amino acid powder (a mixture of 1.25 g adenine sulfate, 0.6 g arginine, 3 g aspartic acid, 3 g glutamic acid, 0.6 g histidine, 0.9 g lysine, 0.6 g methionine, 1.5 g phenylalanine, 11.25 g serine, 0.9 g tyrosine, 4.5 g valine, 6 g threonine and 1.2 g tryptophan). Random one of the selected strains was designated as ARA3-1 strain. This strain can produce arachidonic acid by expressing the Δ12 fatty acid desaturase gene, Δ6 fatty acid desaturase gene, GLELO fatty acid elongase gene, and Δ5 fatty acid desaturase gene from the GAL1/10 promoter upon cultivation in a galactose-containing medium.

後処理

  1. customobtained
  2. customA fragment of about 3.2 kb obtained
  3. additioncontaining, per liter, 6.7 g Yeast nitrogen base

出典と根拠

ORDレコード
ord-094bedc3da33440298a77e80cb720572
出典データセット
uspto-grants-2014_07
特許番号
US08790906B2

反応ノート

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