Isoprene hydrocarbons production upon heterologous transformation of Saccharomyces cerevisiae
Article first published online: 15 MAY 2012
© 2012 The Authors. Journal of Applied Microbiology © 2012 The Society for Applied Microbiology
Journal of Applied Microbiology
Volume 113, Issue 1, pages 52–65, July 2012
How to Cite
Hong, S.-Y., Zurbriggen, A.S. and Melis, A. (2012), Isoprene hydrocarbons production upon heterologous transformation of Saccharomyces cerevisiae. Journal of Applied Microbiology, 113: 52–65. doi: 10.1111/j.1365-2672.2012.05319.x
- Issue published online: 13 JUN 2012
- Article first published online: 15 MAY 2012
- Accepted manuscript online: 21 APR 2012 09:15AM EST
- 2011/2105: received 12 December 2011, revised 1 March 2012 and accepted 3 April 2012
Figure S1 Diagram of the 9.1 kb rDNA repeat unit in S. cerevisiae. The yeast rDNA consists of 100-200 tandemly repeated copies of a 9.1 kb unit on the right arm of chromosome XII. 5S, 5.8S, 18S, and 25S rRNAs are transcribed from the rDNA by RNA polymerase. Each rRNA form is present in a single copy in an 80S yeast ribosome. Abbreviations used: NTS = non-transcribed spacer; ITS = internal transcribed spacer; ETS = external transcribed spacer.
Figure S2 GC-MS analysis of the culture headspace from S. cerevisiae recipient strain and ScT16 transformant. (a) GC and mass spectra of headspace gases generated by the recipient strain S. cerevisiae culture. Upper panel: GC profile of headspace gases from the culture. Lower panel: Mass profile of headspace gases from the same culture. Note the low level (noise level) multiple unspecific lines. (b) GC and mass spectra of headspace vapor generated by S. cerevisiae ScT16 transformant cultures. Upper panel: GC profile of headspace gases from the culture of S. cerevisiae ScT16 transformant. Note the specific peaks not present in the corresponding analysis of the recipient strain. Lower panel: Mass profile of gases from the culture of S. cerevisiae ScT16 transformant. Note the presence of MS lines 53, 67, and 68 from isoprene. Also note the presence of MS line 72 attributed to a variety of products (see Table 1). (c) Mass spectrum lines 53, 67, and 68 from an isoprene standard. (d) 1 ml of gas was withdrawn from the headspace of the sealed flasks and analyzed by GC-FID. Ethanol peaks are shown with a retention time of 5.4 min. Isoprene peaks are shown with a retention time of 6.8 min. Left panel: GC-FID profile of gases from the culture of S. cerevisiae recipient strain. Middle panel: GC-FID profile of gases from the culture of S. cerevisiae ScT16 transformant carrying the SckIspS transgene. (Note: hydroxylated isoprene products overlap with the ethanol peak in this GC analysis.) Right panel: GC-FID profile of an isoprene standard.
Table S1 Primer sequences used in the IspS-containing plasmid construction.
Table S2 Primer sequences used in the yeast colony PCR, Southern hybridization, genomic PCR, and RT-PCR.
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