Production of 10R-hydroxy unsaturated fatty acids from hempseed oil hydrolyzate by recombinant Escherichia coli cells expressing PpoC from Aspergillus nidulans

• Published in: Applied microbiology and biotechnology Vol. 100; no. 18; pp. 7933 - 7944
• Main Authors: Han, Jeong-Eun, Seo, Min-Ju, Shin, Kyung-Chul, Oh, Deok-Kun
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.09.2016; Springer; Springer Nature B.V
• Subjects: Analysis; Aspergillus; Aspergillus nidulans; Aspergillus nidulans - enzymology; Aspergillus nidulans - genetics; Biomedical and Life Sciences; Biotechnological Products and Process Engineering; Biotechnology; Biotransformation; Chemical properties; Dioxygenases - genetics; Dioxygenases - metabolism; E coli; Enzymes; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Essences and essential oils; Fatty acids; Fatty Acids, Unsaturated - metabolism; Fungi; Genetic recombination; Hydrogen-Ion Concentration; Life Sciences; Linoleic Acid - metabolism; Microbial Genetics and Genomics; Microbiological research; Microbiological synthesis; Microbiology; Microorganisms; Physiological aspects; Plant Oils - metabolism; Productivity; Protein expression; Proteins; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Solvents; Studies; Temperature; Unsaturated fatty acids; octadecatrienoic acid; 12; PpoC; octadecadienoic acid; Linoleate 10; 15; dioxygenase; hydroxy-8; Hempseed oil hydrolyzate; 10; Linoleate 10R-dioxygenase; 10R-hydroxy-8E,12Z,15Z-octadecatrienoic acid; Aspergillus nidulans; 10R-hydroxy-8E,12Z-octadecadienoic acid
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-016-7508-6

The first and second preferred substrates of recombinant Escherichia coli cells expressing 10 R -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, 10 R -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 10 R -hydroxy-8 E ,12 Z -octadecadienoic acid (10 R -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 10 R -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 10 R -hydroxy-8 E ,12 Z ,15 Z -octadecatrienoic acid (10 R -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 10 R -HODE and 0.5 g l −1 10 R -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 10 R -hydroxy unsaturated fatty acids.