BackgroundGlucose repression in yeast leads to the sequential or diauxic utilization of mixed sugars and reduces the co-utilization of glucose and xylose from lignocellulosic biomasses. Study of the glucose sensing pathway helps to construct glucose repression-released yeast strains and enhance the utilization of lignocellulosic biomasses.ResultsHerein, the glucose sensor/receptor repressor (SRR) pathway ofKluyveromyces marxianuswhich mainly consisted of KmSnf3, KmGrr1, KmMth1, and KmRgt1 was studied. The disruption ofKmSNF3led to a release of glucose repression, enhanced xylose consumption and did not result in deficient glucose utilization. Over-expression of glucose transporter gene restored the mild decrease of glucose utilization ability ofKmsnf3strain to a similar level of the wildtype strain but did not restore glucose repression. Therefore, the repression on glucose transporter is parallel to glucose repression to xylose and other alternative carbon utilization.KmGRR1disruption also released glucose repression and kept glucose utilization ability, although its xylose utilization ability was very weak with xylose as sole carbon source. The stable mutant of KmMth1-ΔT enabled the release of glucose repression irrespective that the genetic background wasKmsnf3,Kmmth1, or wildtype. Disruption ofKmSNF1in theKmsnf3strain orKmMTH1-ΔToverexpression inKmsnf1strain kept constitutive glucose repression, indicating thatKmSNF1was necessary to release the glucose repression in both SRR and Mig1-Hxk2 pathway. Finally, overexpression ofKmMTH1-ΔTreleased the glucose repression to xylose utilization inS. cerevisiae.ConclusionThe glucose repression-releasedK. marxianusstrains constructed via a modified glucose SRR pathway did not lead to a deficiency in the utilization ability of sugar. The obtained thermotolerant, glucose repression-released, and xylose utilization-enhanced strains are good platforms for the construction of efficient lignocellulosic biomass utilization yeast strains.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12934-023-02138-7.