초록
<P>The oleaginous yeast <I>Trichosporon oleaginosus</I> ATCC 20509 can accumulate up to 70% (w/DCW) triglycerides when cultivated on chemically diverse agricultural or food waste streams. In contrast to other lipogenic yeasts <I>T</I>. <I>oleaginosus</I> is able to efficiently convert constituents of hemicellulose and chitin hydrolysates into lipids. This study focused on establishing the genetic accessibility of <I>T. oleaginosus</I> aimed at manipulating lipid biosynthesis in order to generate high value lipids from waste streams. We demonstrate the first transformation protocol for <I>T. oleaginosus</I> based on <I>Agrobacterium tumefaciens</I>. Strong heterologous gene expression of a codon optimized YFP reporter protein was achieved using the constitutive promotor from the endogenous glyceraldehyde-3-phosphate dehydrogenase gene. Subsequently, we evaluated the ability of <I>T. oleaginosus</I> to generate non-natural fatty acid profiles by heterologous expression of several fatty acid modifying enzymes. <I>De novo</I> lipid generation of these recombinant strains was evaluated on diverse carbon sources. Compared to the wild type, recombinant yeast strains showed an increase of α-linolenic acid production from 2.8% to 21% with respect to the total cellular fatty acid content (TFA). Further, we designed yeast strains able to generate the non-native, polyunsaturated very long chain fatty acids eicosatrienoic (16% TFA) and eicosadienoic acid (9% TFA), respectively. Alternatively, <I>T. oleaginosus</I> was engineered to produce the non-native (<I>E</I>-10, <I>Z</I>-12) conjugated linoleic acid, which was generated up to 2.6% TFA. This work demonstrates, that <I>T. oleaginosus</I> ATCC 20509 can be used as versatile biotechnology platform to transform industrial waste streams into designed, high value fatty acids.</P><BR><BR><P>Graphic Abstract</P><P>Establishing genetic engineering tools for sustainable production of tailor made fatty acids in the non-conventional, oleaginous yeast <I>Trichosporon oleaginosus</I>.<BR><IMG SRC='http://pubs.rsc.org/services/images/RSCpubs.ePlatform.Service.FreeContent.ImageService.svc/ImageService/image/GA?id=c5gc01767j'><BR></P>