title: Production of 10R-hydroxy unsaturated fatty acids from hempseed oil hydrolyzate by recombinant Escherichia coli cells expressing PpoC from Aspergillus nidulans.
abstract: The first and second preferred substrates of recombinant Escherichia coli cells expressing 10R-dioxygenase (PpoC) from Aspergillus nidulans and the purified enzyme were linoleic acid and α-linolenic acid, respectively. PpoC in cells showed higher thermal and reaction stabilities compared to purified PpoC. Thus, 10R-hydroxy unsaturated fatty acids were produced from linoleic acid, α-linolenic acid, and hempseed oil hydrolyzate containing linoleic acid and α-linolenic acid as substrates by whole recombinant cells expressing PpoC. The optimal reaction conditions for the production of 10R-hydroxy-8E,12Z-octadecadienoic acid (10R-HODE) were pH 8.0, 30 °C, 250 rpm, 5 % (v/v) dimethyl sulfoxide, 5 g l(-1) linoleic acid, and 60 g l(-1) cells in 100-ml baffled flask. Under these conditions, whole recombinant cells expressing PpoC produced 2.7 g l(-1) 10R-HODE from 5 g l(-1) linoleic acid for 40 min, with a conversion yield of 54 % (w/w) and a productivity of 4.0 g l(-1) h(-1); produced 2.2 g l(-1) 10R-hydroxy-8E,12Z,15Z-octadecatrienoic acid (10R-HOTrE) from 3 g l(-1) α-linolenic acid for 30 min, with a conversion yield of 72 % (w/w) and a productivity of 4.3 g l(-1) h(-1); and produced 1.8 g l(-1) 10R-HODE and 0.5 g l(-1) 10R-HOTrE from 5 g l(-1) hempseed oil hydrolyzate containing 2.5 g l(-1) linoleic acid and 1.0 g l(-1) α-linolenic acid for 30 min, with a conversion yield of 74 and 51 % (w/w), respectively, and a productivity of 3.6 and 1.0 g l(-1) h(-1), respectively. To the best of our knowledge, this is the first report on the biotechnological production of 10R-hydroxy unsaturated fatty acids.
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University, Seoul, 05029, Republic of Korea.
authors.forename: Jeong-Eun
authors.initials: JE
authors.lastname: Han
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University, Seoul, 05029, Republic of Korea.
authors.forename: Min-Ju
authors.initials: MJ
authors.lastname: Seo
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University, Seoul, 05029, Republic of Korea.
authors.forename: Kyung-Chul
authors.initials: KC
authors.lastname: Shin
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University, Seoul, 05029, Republic of Korea. deokkun@konkuk.ac.kr.
authors.forename: Deok-Kun
authors.initials: DK
authors.lastname: Oh
mesh_terms: D001233:Aspergillus nidulans / Q000201:enzymology* / Q000235:genetics; D001711:Biotransformation; D049308:Dioxygenases / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005231:Fatty Acids, Unsaturated / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D019787:Linoleic Acid / Q000378:metabolism; D010938:Plant Oils / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D013696:Temperature
chemical_list: D005231:Fatty Acids, Unsaturated; D010938:Plant Oils; D011994:Recombinant Proteins; D019787:Linoleic Acid; D049308:Dioxygenases
keywords: 10R-hydroxy-8E,12Z,15Z-octadecatrienoic acid; 10R-hydroxy-8E,12Z-octadecadienoic acid; Aspergillus nidulans; Hempseed oil hydrolyzate; Linoleate 10R-dioxygenase; PpoC