Abstract
This chapter gives an overview of the “minimum genome factory” (MGF) of the fission yeast Schizosaccharomyces pombe (S. pombe). The S. pombe genome is one of the smallest found in free-living eukaryotes. We engineered a reduction in the number of S. pombe genes using a large-scale gene deletion method called the LATOUR method. This method enabled us to identify the minimum gene set required for growth under laboratory conditions. The genome-reduced strain has four deleted regions: 168.4 kb of the left arm of chromosome I; 155.4 kb of the right arm of chromosome I; 211.7 kb of the left arm of chromosome II; and 121.6 kb of the right arm of chromosome II. These changes represent a loss of 223 genes of an estimated 5,100. The 657.3-kb deletion strain was less efficient at taking up glucose and some amino acids from the growth media than the parental strain. This strain also showed increased gene expression of the mating pheromone M-factor precursor and NADP-specific glutamate dehydrogenase. There was also a 2.7-fold increase in the concentration of cellular ATP, whereas levels of heterologously produced proteins, such as the green fluorescent protein and the secreted human growth hormone, increased by 1.7 fold and 1.8 fold, respectively.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Ara K, Ozaki K, Nakamura K et al (2007) Bacillus minimum genome factory: effective utilization of microbial genome information. Biotechnol Appl Biochem 46(pt 3):169–178
Baba T, Ara T, Hasegawa M et al (2006) Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection. Mol Syst Biol 2:20060008. doi:2010.1038/msb4100050
Decottignies A, Sanchez-Perez I, Nurse P (2003) Schizosaccharomyces pombe essential genes: a pilot study. Genome Res 13(3):399–406
Fantes P, Nurse P (1977) Control of cell size at division in fission yeast by a growth-modulated size control over nuclear division. Exp Cell Res 107(2):377–386
Fujio T (2007) Minimum genome factory: innovation in bioprocesses through genome science. Biotechnol Appl Biochem 46(pt 3):145–146
Giga-Hama Y, Tohda H, Takegawa K et al (2007) Schizosaccharomyces pombe minimum genome factory. Biotechnol Appl Biochem 46(Pt 3):147–155
Hirashima K, Iwaki T, Takegawa K et al (2006) A simple and effective chromosome modification method for large-scale deletion of genome sequences and identification of essential genes in fission yeast. Nucleic Acids Res 34(2):e11. doi:10.1093/nar/gnj1011
Idiris A, Tohda H, Bi KW et al (2006) Enhanced productivity of protease-sensitive heterologous proteins by disruption of multiple protease genes in the fission yeast Schizosaccharomyces pombe. Appl Microbiol Biotechnol 73(2):404–420
Idiris A, Tohda H, Sasaki M et al (2009) Enhanced protein secretion from multiprotease-deficient fission yeast by modification of its vacuolar protein sorting pathway. Appl Microbiol Biotechnol 85(3):667–677
Idiris A, Tohda H, Kumagai H et al (2010) Engineering of protein secretion in yeast: strategies and impact on protein production. Appl Microbiol Biotechnol 86(2):403–417
Kim DU, Hayles J, Kim D et al (2010) Analysis of a genome-wide set of gene deletions in the fission yeast Schizosaccharomyces pombe. Nat Biotechnol 28(6):617–623
Kobayashi K, Ehrlich SD, Albertini A et al (2003) Essential Bacillus subtilis genes. Proc Natl Acad Sci USA 100(8):4678–4683
Manabe K, Kageyama Y, Morimoto T et al (2011) Combined effect of improved cell yield and increased specific productivity enhances recombinant enzyme production in genome-reduced Bacillus subtilis strain MGB874. Appl Environ Microbiol 77(23):8370–8381
Matsuo T, Otsubo Y, Urano J et al (2007) Loss of the TOR kinase Tor2 mimics nitrogen starvation and activates the sexual development pathway in fission yeast. Mol Cell Biol 27(8):3154–3164
Medema MH, van Raaphorst R, Takano E et al (2012) Computational tools for the synthetic design of biochemical pathways. Nat Rev Microbiol 10(3):191–202
Mizoguchi H, Mori H, Fujio T (2007) Escherichia coli minimum genome factory. Biotechnol Appl Biochem 46(Pt 3):157–167
Mizoguchi H, Sawano Y, Kato J et al (2008) Superpositioning of deletions promotes growth of Escherichia coli with a reduced genome. DNA Res 15(5):277–284
Morimoto T, Kadoya R, Endo K et al (2008) Enhanced recombinant protein productivity by genome reduction in Bacillus subtilis. DNA Res 15(2):73–81
Moya A, Gil R, Latorre A et al (2009) Toward minimal bacterial cells: evolution vs. design. FEMS Microbiol Rev 33(1):225–235
Murakami K, Tao E, Ito Y et al (2007) Large scale deletions in the Saccharomyces cerevisiae genome create strains with altered regulation of carbon metabolism. Appl Microbiol Biotechnol 75(3):589–597
Posfai G, Plunkett G 3rd, Feher T et al (2006) Emergent properties of reduced-genome Escherichia coli. Science 312(5776):1044–1046
Sasaki M, Kumagai H, Takegawa K et al (2013) Characterization of genome-reduced fission yeast strains. Nucleic Acids Res 41(10):5382–5399
Wood V, Gwilliam R, Rajandream MA et al (2002) The genome sequence of Schizosaccharomyces pombe. Nature (Lond) 415(6874):871–880
Zhao Y, Lieberman HB (1995) Schizosaccharomyces pombe: a model for molecular studies of eukaryotic genes. DNA Cell Biol 14(5):359–371
Acknowledgments
This study was partly supported by the Ministry of Economy, Trade and Industry (Project for the Development of a Technological Infrastructure for Industrial Bioprocesses on Research and Development of New Industrial Science and Technology Frontiers supported by New Energy and Industrial Technology Development Organization). We are sincerely grateful to the late Yuko Giga-Hama.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Japan
About this chapter
Cite this chapter
Kumagai, H., Sasaki, M., Idiris, A., Tohda, H. (2014). Minimum Genome Factories in Schizosaccharomyces pombe . In: Anazawa, H., Shimizu, S. (eds) Microbial Production. Springer, Tokyo. https://doi.org/10.1007/978-4-431-54607-8_2
Download citation
DOI: https://doi.org/10.1007/978-4-431-54607-8_2
Published:
Publisher Name: Springer, Tokyo
Print ISBN: 978-4-431-54606-1
Online ISBN: 978-4-431-54607-8
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)