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

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• 作者：Jeong-Eun Han ; Min-Ju Seo ; Kyung-Chul Shin…
• 刊名：Applied Microbiology and Biotechnology
• 出版年：2016
• 出版时间：September 2016
• 年：2016
• 卷：100
• 期：18
• 页码：7933-7944
• 全文大小：591 KB
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Biotechnology
  Microbiology
  Microbial Genetics and Genomics
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-0614
• 卷排序：100

文摘

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−1α-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−1α-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.KeywordsAspergillus nidulansLinoleate 10R-dioxygenasePpoCHempseed oil hydrolyzate10R-hydroxy-8E,12Z-octadecadienoic acid10R-hydroxy-8E,12Z,15Z-octadecatrienoic acid