title: Heterologous expression of xylanase enzymes in lipogenic yeast Yarrowia lipolytica.
abstract: To develop a direct microbial sugar conversion platform for the production of lipids, drop-in fuels and chemicals from cellulosic biomass substrate, we chose Yarrowia lipolytica as a viable demonstration strain. Y. lipolytica is known to accumulate lipids intracellularly and is capable of metabolizing sugars to produce lipids; however, it lacks the lignocellulose-degrading enzymes needed to break down biomass directly. While research is continuing on the development of a Y. lipolytica strain able to degrade cellulose, in this study, we present successful expression of several xylanases in Y. lipolytica. The XynII and XlnD expressing Yarrowia strains exhibited an ability to grow on xylan mineral plates. This was shown by Congo Red staining of halo zones on xylan mineral plates. Enzymatic activity tests further demonstrated active expression of XynII and XlnD in Y. lipolytica. Furthermore, synergistic action in converting xylan to xylose was observed when XlnD acted in concert with XynII. The successful expression of these xylanases in Yarrowia further advances us toward our goal to develop a direct microbial conversion process using this organism.
authors.affiliation: Biosciences Center, National Renewable Energy Laboratory, Golden, Colorado, United States of America.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.affiliation: Biosciences Center, National Renewable Energy Laboratory, Golden, Colorado, United States of America.
authors.forename: Hui
authors.initials: H
authors.lastname: Wei
authors.affiliation: Biosciences Center, National Renewable Energy Laboratory, Golden, Colorado, United States of America.
authors.forename: Markus
authors.initials: M
authors.lastname: Alahuhta
authors.affiliation: National Bioenergy Center, National Renewable Energy Laboratory, Golden, Colorado, United States of America.
authors.forename: Xiaowen
authors.initials: X
authors.lastname: Chen
authors.affiliation: National Bioenergy Center, National Renewable Energy Laboratory, Golden, Colorado, United States of America.
authors.forename: Deborah
authors.initials: D
authors.lastname: Hyman
authors.affiliation: Biosciences Center, National Renewable Energy Laboratory, Golden, Colorado, United States of America.
authors.forename: David K
authors.initials: DK
authors.lastname: Johnson
authors.affiliation: National Bioenergy Center, National Renewable Energy Laboratory, Golden, Colorado, United States of America.
authors.forename: Min
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Biosciences Center, National Renewable Energy Laboratory, Golden, Colorado, United States of America.
authors.forename: Michael E
authors.initials: ME
authors.lastname: Himmel
mesh_terms: D018533:Biomass; D002482:Cellulose / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D043364:Endo-1,4-beta Xylanases / Q000096:biosynthesis*; D020869:Gene Expression Profiling; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D007218:Industrial Microbiology; D008031:Lignin / Q000737:chemistry; D013058:Mass Spectrometry; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis; D014990:Xylans / Q000737:chemistry; D014994:Xylose / Q000737:chemistry; D025062:Yarrowia / Q000201:enzymology*; D003313:Zea mays / Q000737:chemistry
chemical_list: D011994:Recombinant Proteins; D014990:Xylans; C036909:lignocellulose; D002482:Cellulose; D008031:Lignin; D014994:Xylose; D043364:Endo-1,4-beta Xylanases