title: Expression of bacterial levanase in yeast enables simultaneous saccharification and fermentation of grass juice to bioethanol.
abstract: This study demonstrates use of recombinant yeast to simultaneously saccharify and ferment grass juice (GJ) to bioethanol. A modified Bacillus subtilis levanase gene (sacC) in which the native bacterial signal sequence was replaced with a yeast α-factor domain, was synthesised with yeast codon preferences and transformed into Saccharomyces cerevisiae (strain AH22) using the expression vector pMA91. AH22:psacC transformants secreted sacCp as an active, hyper-glycosylated (>180 kDa) protein allowing them to utilise inulin (β[2-1] linked fructose) and levan (β[2-6] linkages) as growth substrates. The control (AH22:pMA91) strain, transformed with empty plasmid DNA was not able to utilise inulin or levan. When cultured on untreated GJ levels of growth and bioethanol production were significantly higher in experiments with AH22:psacC than with AH22:pMA91. Bioethanol yields from AH22:psacC grown on GJ (32.7[±4] mg mL(-1)) compared closely to those recently achieved (Martel et al., 2010) using enzymatically pre-hydrolysed GJ (36.8[±4] mg mL(-1)).
authors.affiliation: Institute of Life Science and School of Medicine, Swansea University, Swansea SA2 8PP, Wales, UK.
authors.forename: C M
authors.initials: CM
authors.lastname: Martel
authors.forename: J E
authors.initials: JE
authors.lastname: Parker
authors.forename: C J
authors.initials: CJ
authors.lastname: Jackson
authors.forename: A G S
authors.initials: AG
authors.lastname: Warrilow
authors.forename: N
authors.initials: N
authors.lastname: Rolley
authors.forename: C
authors.initials: C
authors.lastname: Greig
authors.forename: S M
authors.initials: SM
authors.lastname: Morris
authors.forename: I S
authors.initials: IS
authors.lastname: Donnison
authors.forename: D E
authors.initials: DE
authors.lastname: Kelly
authors.forename: S L
authors.initials: SL
authors.lastname: Kelly
mesh_terms: D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry; D056804:Biofuels / Q000032:analysis*; D001709:Biotechnology; D050260:Carbohydrate Metabolism; D000431:Ethanol / Q000138:chemical synthesis*; D005285:Fermentation; D006026:Glycoside Hydrolases / Q000378:metabolism*; D008969:Molecular Sequence Data; D006109:Poaceae / Q000378:metabolism*; D011995:Recombination, Genetic / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D014170:Transformation, Genetic
chemical_list: D001426:Bacterial Proteins; D056804:Biofuels; D000431:Ethanol; D006026:Glycoside Hydrolases; C021512:levanase