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Applied Microbiology and Biotechnology

, Volume 103, Issue 1, pp 395–410 | Cite as

Functional analysis of Mig1 and Rag5 as expressional regulators in thermotolerant yeast Kluyveromyces marxianus

  • Mochamad Nurcholis
  • Sukanya Nitiyon
  • Suprayogi
  • Nadchanok Rodrussamee
  • Noppon Lertwattanasakul
  • Savitree Limtong
  • Tomoyuki Kosaka
  • Mamoru YamadaEmail author
Applied microbial and cell physiology
  • 254 Downloads

Abstract

To analyze the glucose repression mechanism in the thermotolerant yeast Kluyveromyces marxianus, disrupted mutants of genes for Mig1 and Rag5 as orthologs of Mig1 and Hxk2, respectively, in Saccharomyces cerevisiae were constructed, and their characteristics were compared with those of the corresponding mutants of S. cerevisiae. MIG1 mutants of both yeasts exhibited more resistance than the corresponding parental strains to 2-deoxyglucose (2-DOG). Histidine was found to be essential for the growth of Kmmig1, but not that of Kmrag5, suggesting that MIG1 is required for histidine biosynthesis in K. marxianus. Moreover, Kmrag5 and Schxk2 were more resistant than the corresponding MIG1 mutant to 2-DOG, and only the latter increased the utilization speed of sucrose in the presence of glucose. Kmrag5 exhibited very low activities for gluco-hexokinase and hexokinase and, unlike Schxk2, showed very slow growth and a low level of ethanol production in a glucose medium. Furthermore, Kmrag5, but not Kmmig1, exhibited high inulinase activity in a glucose medium and exhibited greatly delayed utilization of accumulated fructose in the medium containing both glucose and sucrose. Transcription analysis revealed that the expression levels of INU1 for inulinase and GLK1 for glucokinase in Kmrag5 were higher than those in the parental strain; the expression level of INU1 in Kmmig1 was higher, but the expression levels of RAG1 for a low-affinity glucose transporter in Kmmig1 and Kmrag5 were lower. These findings suggest that except for regulation of histidine biosynthesis, Mig1 and Rag5 of K. marxianus play similar roles in the regulation of gene expression and share some functions with Mig1 and Hxk2, respectively, in S. cerevisiae.

Keywords

Kluyveromyces marxianus Glucose repression MIG1 RAG5 Histidine biosynthesis 

Notes

Acknowledgments

We thank K. Matsushita and T. Yakushi for their helpful discussion.

Authors’ contributions

MN obtained and carried out characterization of MIG1 and RAG5 disrupted mutants and performed enzyme assay, gene expression, and writing the manuscript. SN was involved in construction of MIG1 and RAG5 disrupted mutants. S was involved in obtaining mutant no. 23. SL isolated K. marxianus DMKU 3-1042. NR, NL, and TK were participated in discussion of the study. MY was contributed in the experimental design and discussion for writing the manuscript. All authors read and approved the final manuscript.

Funding information

The Govermment of Indonesia gave financial support (to M. N) through BPPLN-DIKTI Scholarship, Ministry of Research, Technology, and Higher Education. This work was supported by The Core to Core Program A. Advanced Research Networks, which was granted by the Japan Society for the Promotion of Science, the National Research Council of Thailand, Ministry of Science and Technology in Vietnam, National Univ. of Laos, Univ. of Brawijaya, and Beuth Univ. of Applied Science Berlin and was also supported by the Program for Promotion of Basic Research Activities for Innovative Biosciences, NEDO, Special coordination and by Japan Science and Technology Agency, Ministry of Research, Technology and Higher Education of the Republic of Indonesia, Agricultural Research Development Agency of Thailand and Ministry of Science and Technology of Laos as part of the e-ASIA Joint Research Program (e-ASIA JRP).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical statement

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2018_9462_MOESM1_ESM.pdf (540 kb)
ESM 1 (PDF 539 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Mochamad Nurcholis
    • 1
  • Sukanya Nitiyon
    • 1
  • Suprayogi
    • 2
  • Nadchanok Rodrussamee
    • 3
  • Noppon Lertwattanasakul
    • 4
  • Savitree Limtong
    • 4
  • Tomoyuki Kosaka
    • 5
    • 6
    • 7
  • Mamoru Yamada
    • 1
    • 5
    • 6
    • 7
    Email author
  1. 1.Graduate School of MedicineYamaguchi UniversityUbeJapan
  2. 2.Department of Agroindustrial Technology, Faculty of Agricultural TechnologyBrawijaya UniversityMalangIndonesia
  3. 3.Department of Biology, Faculty of ScienceChiang Mai UniversityChiang MaiThailand
  4. 4.Department of Microbiology, Faculty of ScienceKasetsart UniversityBangkokThailand
  5. 5.Department of Biological Chemistry, Faculty of AgricultureYamaguchi UniversityYamaguchiJapan
  6. 6.Graduate School of Science and Technology for InnovationYamaguchi UniversityYamaguchiJapan
  7. 7.Research Center for Thermotolerant Microbial ResourcesYamaguchi UniversityYamaguchiJapan

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