Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling by array of Saccharomyces cerevisiae strain VL3 with or without the the co-inoculation of a second yeast species, Pichia kluyveri (PKKR1).

ABSTRACT: Replicate cultres of Saccharomyces cerevisiae strain VL3 (a commercial wine strain) were inititaed with and without the the co-inoculation of a second yeast species, Pichia kluyveri (PKKR1). In addition replicate cultures of Pichia kluyveri were initiated. The media for all cultures was sterile Sauvignon blanc grape juice. Triplicates of each treatment were destructively harvested at days 2, 9 and 16 of fermentation and RNA extracted. RNA from S. cerevisiae could not be preferentially extracted from co-ferments. P. kluyveri genome has not been sequenced and so homology to S. cerevisiae probes on the array could not be accurately estimated. We extracted RNA from the P. kluyveri single ferments and hybridised them to the S. cerevisiae arrays to control for this. These species diverged over 100 MYA, and we did not observe abundant homology by way of cross hybridisation. The mean probe intensity for P. kluyveri cDNA was 17-fold lower than for S. cerevisiae, but some probes reported reasonable signal. The intensity distribution of these probes fit a log-normal distribution (P<0.01). 97% of the distribution fell below one-thirtieth of the S. cerevisiae maximum intensity: the remaining 3%, comprising 429 probes, potentially have reasonable homology to P. kluyveri cDNA and we removed these from all analyses. Please see associated additional file 'removePKKR1.r' R code to achieve this.

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Matthew Goddard

PROVIDER: E-MTAB-1235 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Over the past three decades, due to the universal application of Pichia yeast in the fermentation industry as well as the establishment of Pichia pastoris fermentation process for more than 30 years, the technology of the whole process has become very mature and has now reached a stagnated period of growth. However, studies and research conducted on the genomics of the classic fermentation process and the uncovering of the biological phenomena in the fermentation process from the point of view of high-throughput gene or protein is still in its early stages, and there is still insufficient data within this field. First, in collaboration with Agilent Company, we designed and prepared an expression microarray that could be used for the detection of Pichia pastoris transcriptomics. The transcriptomic changes in the five key technology steps (time points), during the fermentation process of Pichia pastoris would then be detected with the aid of an expression microarray. The five key steps of technology described above formed two important biological processes, namely, the limiting carbon source replacement and secondly, the fermentative production of exogenous proteins. The biological phenomena involved in these two processes were displayed and analyzed at the transcriptional level. In addition to this, with regard to the most important function in the fermentation process of Pichia pastoris, oxid-reduction, the metabolic drift process was analyzed and the important genes that might dominate the changes in the metabolic flux were discovered creatively by using the function tree method in this paper. This study was undertaken from the point of view of the transcriptome and the biological phenomena in the fermemntation process of Pichia pastoris. Both of which, were thoroughly explained during this study. The hope is for many more researchers to optimize the strain fermentation process, to produce proteins at the genetic level, as well as providing and obtaining new perspectives and detailed scientific data for the continued development within this field. The transcriptomic changes in the five key technology steps (time points), during the fermentation process of Pichia pastoris would then be detected with the aid of an expression microarray.

Dataset Information

Transcription profiling by array of Saccharomyces cerevisiae strain VL3 with or without the the co-inoculation of a second yeast species, Pichia kluyveri (PKKR1).

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