The Open Biotechnology Journal

ISSN: 1874-0707 ― Volume 14, 2020

Characteristics of kanMX4-inserted Mutants that Exhibit 2-Deoxyglucose Resistance in Thermotolerant Yeast Kluyveromyces marxianus

Suprayogi1, M. Nurcholis1, M. Murata1, N. Lertwattanasakul2, T. Kosaka4, N. Rodrussamee1, 3, S. Limtong2, M. Yamada1, 4, *
1 Applied Molecular Bioscience, Graduate School of Medicine, Yamaguchi University, Ube, 755-8505, Japan
2 Department of Microbiology, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand
3 Faculty of Science, Chiangmai University, Chiangmai 50200, Thailand
4 Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi, 753-8515, Japan


Kluyveromyces marxianus has the attractive potential of utilization capability of various sugars in addition to thermal tolerance and protein productivity. The yeast, however, has an intrinsic system for catabolite repression, by which cells down-regulate the metabolism of alternative sugars when glucose coexists. To acquire glucose-repression-free mutants, we isolated and characterized 2-deoxyglucose-resistant mutants from kanMX4-inserted mutants. The insertion site was determined by TAIL-PCR followed by nucleotide sequencing, indicating that the kanMX4 cassette was intragenically or intergenically inserted. Further analysis of the sugar utilization ability allowed to classify the intragenically inserted mutants including the mig1 mutant into two categories. One group showed enhanced utilization of xylose in the presense of glucose, presumably due to a defect in the glucose-repression mechanism, and the other group showed delayed utilization of glucose, probably by reduction of the uptake or initial catabolism of glucose. Considering the possible functions of the disrupted genes in these mutants, it is assumed that K. marxianus has undiscovered mechanisms for glucose repression and complex regulation for the uptake or initial catabolism of glucose.

Keywords: Glucose repression, 2-deoxyglucose-resistant mutants, Kluyveromyces marxianus, kanMX4 insertion, xylose fermentation.

Article Information

Identifiers and Pagination:

Year: 2016
Volume: 10
First Page: 208
Last Page: 222
Publisher Id: TOBIOTJ-10-208
DOI: 10.2174/18740707016100100208

Article History:

Received Date: 17/10/2015
Revision Received Date: 31/12/2015
Acceptance Date: 3/2/2016
Electronic publication date: 06/05/2016
Collection year: 2015

© Suprayogi et al.; Licensee Bentham Open.

open-access license: This is an open access article licensed under the terms of the Creative Commons Attribution-Non-Commercial 4.0 International Public License (CC BY-NC 4.0) (, which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.

* Address correspondence to this author at the Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi, 753-8515, Japan; E-mail:

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