Metabolic engineering of fatty alcohol production in transgenic hairy roots of Crambe abyssinica

• Published in: Biotechnology and bioengineering 2017-06, Vol.114 (6), p.1275-1282
• Main Authors: Miklaszewska, Magdalena, Banaś, Antoni, Królicka, Aleksandra
• Format: Article
• Language: English
• Subjects: Agrobacterium; Alcohols; Aldehyde Oxidoreductases - genetics; Aldehyde Oxidoreductases - metabolism; Arabidopsis thaliana; Bioengineering; Crambe; Crambe abyssinica; Crambe Plant - physiology; fatty acyl reductase; fatty alcohols; Fatty Alcohols - isolation & purification; Fatty Alcohols - metabolism; Genes; hairy roots; Metabolic Engineering - methods; Plant Roots - physiology; Plants, Genetically Modified - genetics; Plants, Genetically Modified - metabolism; Receiving; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Reductases; Roots; Simmondsia chinensis; Transformations; Transgenic
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.26234

ABSTRACT Biotechnological production of fatty alcohols, important raw materials in the chemical industry, has been receiving considerable attention in recent years. Fatty alcohols are formed by the reduction of fatty acyl‐CoAs or fatty acyl‐ACPs catalyzed by a fatty acyl reductase (FAR). In this study, we introduced genes encoding FARs from Arabidopsis thaliana (AtFAR5) and Simmondsia chinensis (ScFAR) into Crambe abyssinica hairy roots via Agrobacterium rhizogenes‐mediated transformation. The efficiency of the transformation ranged between 30 and 45%. The fatty alcohols were only detected in the transgenic hairy root lines expressing ScFAR gene. In all tested lines stearyl alcohol (18:0‐OH), arachidyl alcohol (20:0‐OH), and behenyl alcohol (22:0‐OH) were produced. The content of 18:0‐OH varied from 1 to 3% of total fatty acids and fatty alcohols, while the amount of either 20:0‐OH and 22:0‐OH did not exceed 2%. The transgenic hairy root lines produced from 0.98 to 2.59 nmol of fatty alcohols per mg of dry weight. Very low activity of ScFAR was detected in the microsomal fractions isolated from the selected hairy root lines. To our knowledge, this is the first report on the fatty alcohol production in the hairy root cultures. Biotechnol. Bioeng. 2017;114: 1275–1282. © 2016 Wiley Periodicals, Inc. The hairy roots of Crambe abyssinica have been metabolically engineered to produce fatty alcohols. The introduction of the fatty acyl‐CoA reductase gene from Simmondsia chinensis via Agrobacterium‐mediated transformation resulted in the establishment of the hairy roots lines synthesizing stearyl alcohol (18:0‐OH), arachidyl alcohol (20:0‐OH), and behenyl alcohol (22:0‐OH).