Ethanol production from xylose is highly increased by the Kluyveromyces marxianus mutant 17694-DH1

Kwon, Deok-Ho; Park, Jae-Bum; Hong, Eunsoo; Ha, Suk‐Jin

doi:10.1007/s00449-018-2014-0

Cited by 14 publications

(15 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In previous studies, 2-DG-resistant mutants demonstrated simultaneous glucose and xylose cofermentation capabilities following relief of glucose catabolite repression [12, 13, 23]. A laboratory-directed evolutionary approach was therefore applied to K. marxianus 17694-DH1 to alleviate glucose catabolite repression using 2-DG [22]. Pre-cultured cells were initially inoculated into YP media (10 g/L yeast extract, 20 g/L bacto peptone) containing 15 g/L 2-DG and 40 g/L xylose (2-DG/xylose media).…”

Section: Resultsmentioning

confidence: 99%

“…Kluyveromyces marxianus 17694-DH1, capable of fermenting xylose to ethanol, was used as the parental strain in a directed evolutionary approach [22]. For yeast inoculum preparation, pre-culture was carried out in YP media with 20 g/L glucose (YPD 20 ) or 20 g/L xylose (YPX 20 ).…”

Section: Methodsmentioning

confidence: 99%

“…Previously, the K. marxianus 17694-DH1 mutant was isolated following a random mutagenesis/directed evolutionary approach and shown to be effective for ethanol production from xylose [22]. However, K. marxianus 17694-DH1 displays strong glucose catabolite repression when using glucose and xylose as mixed sugars.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Alleviation of catabolite repression in Kluyveromyces marxianus: the thermotolerant SBK1 mutant simultaneously coferments glucose and xylose

Kim

Kwon

Park

et al. 2019

Biotechnol Biofuels

Self Cite

View full text Add to dashboard Cite

Background Simultaneous cofermentation of glucose and xylose mixtures would be a cost-effective solution for the conversion of cellulosic biomass to high-value products. However, most yeasts ferment glucose and xylose sequentially due to glucose catabolite repression. A well known thermotolerant yeast, Kluyveromyces marxianus , was selected for this work because it possesses cost-effective advantages over Saccharomyces cerevisiae for biofuel production from cellulosic biomass. Results In the present study, we employed a directed evolutionary approach using 2-deoxyglucose to develop a thermotolerant mutant capable of simultaneous cofermentation of glucose and xylose by alleviating catabolite repression. The selected mutant, K. marxianus SBK1, simultaneously cofermented 40 g/L glucose and 28 g/L xylose to produce 23.82 g/L ethanol at 40 °C. This outcome corresponded to a yield of 0.35 g/g and productivity of 0.33 g/L h, representing an 84% and 129% improvement, respectively, over the parental strain. Interestingly, following mutagenesis the overall transcriptome of the glycolysis pathway was highly downregulated in K. marxianus SBK1, except for glucokinase-1 (GLK1) which was 21-fold upregulated. Amino acid sequence of GLK1 from K. marxianus SBK1 revealed three amino acid mutations which led to more than 22-fold lower enzymatic activity compared to the parental strain. Conclusions We herein successfully demonstrated that the cofermentation of a sugar mixture is a promising strategy for the efficient utilization of cellulosic biomass by K. marxianus SBK1. Through introduction of additional biosynthetic pathways, K. marxianus SBK1 could become a chassis-type strain for the production of fuels and chemicals from cellulosic biomass. Electronic supplementary material The online version of this article (10.1186/s13068-019-1431-x) contains supplementary material, which is available to authorized users.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Alleviation of catabolite repression in Kluyveromyces marxianus: the thermotolerant SBK1 mutant simultaneously coferments glucose and xylose

Kim

Kwon

Park

et al. 2019

Biotechnol Biofuels

Self Cite

View full text Add to dashboard Cite

show abstract

“…K. marxianus can even grow at 52 °C due to its notable thermotolerance [9], while S. cerevisiae has optimum temperatures ranging between 30 and 37 °C [10, 11]. Also, besides glucose, K. marxianus is able to utilize a variety of carbon sources, including inulin, xylose, and lactose [12, 13], which cannot be used by S. cerevisiae [9]. Furthermore, ethanol generation prefers an anaerobic environment; thus, the capability to grow under full anaerobiosis is also required for a yeast strain to be used in a fuel ethanol-producing process.…”

Section: Introductionmentioning

confidence: 99%

Kluyveromyces marxianus developing ethanol tolerance during adaptive evolution with significant improvements of multiple pathways

Wang

Zhan

et al. 2019

Biotechnol Biofuels

View full text Add to dashboard Cite

BackgroundKluyveromyces marxianus, the known fastest-growing eukaryote on the earth, has remarkable thermotolerance and capacity to utilize various agricultural residues to produce low-cost bioethanol, and hence is industrially important to resolve the imminent energy shortage crisis. Currently, the poor ethanol tolerance hinders its operable application in the industry, and it is necessary to improve K. marxianus’ ethanol resistance and unravel the underlying systematical mechanisms. However, this has been seldom reported to date.ResultsWe carried out a wild-type haploid K. marxianus FIM1 in adaptive evolution in 6% (v/v) ethanol. After 100-day evolution, the KM-100d population was obtained; its ethanol tolerance increased up to 10% (v/v). Interestingly, DNA analysis and RNA-seq analysis showed that KM-100d yeasts’ ethanol tolerance improvement was not due to ploidy change or meaningful mutations, but founded on transcriptional reprogramming in a genome-wide range. Even growth in an ethanol-free medium, many genes in KM-100d maintained their up-regulation. Especially, pathways of ethanol consumption, membrane lipid biosynthesis, anti-osmotic pressure, anti-oxidative stress, and protein folding were generally up-regulated in KM-100d to resist ethanol. Notably, enhancement of the secretory pathway may be the new strategy KM-100d developed to anti-osmotic pressure, instead of the traditional glycerol production way in S. cerevisiae. Inferred from the transcriptome data, besides ethanol tolerance, KM-100d may also develop the ability to resist osmotic, oxidative, and thermic stresses, and this was further confirmed by the cell viability test. Furthermore, under such environmental stresses, KM-100d greatly improved ethanol production than the original strain. In addition, we found that K. marxianus may adopt distinct routes to resist different ethanol concentrations. Trehalose biosynthesis was required for low ethanol, while sterol biosynthesis and the whole secretory pathway were activated for high ethanol.ConclusionsThis study reveals that ethanol-driven laboratory evolution could improve K. marxianus’ ethanol tolerance via significant up-regulation of multiple pathways including anti-osmotic, anti-oxidative, and anti-thermic processes, and indeed consequently raised ethanol yield in industrial high-temperature and high-ethanol circumstance. Our findings give genetic clues for further rational optimization of K. marxianus’ ethanol production, and also partly confirm the positively correlated relationship between yeast’s ethanol tolerance and production.Electronic supplementary materialThe online version of this article (10.1186/s13068-019-1393-z) contains supplementary material, which is available to authorized users.

show abstract

“…To date, all studies that addressed gene expression in this yeast focused on transcriptional effects via RNA-Seq experiments. Aspects that have been studied include growth and ethanol production on alternative sugar substrates such as xylose (Schabort et al 2016;Kwon et al 2019) and inulin (Gao et al 2015); ethanol tolerance during adaptive laboratory evolution (Mo et al 2019); response to growth inhibitors derived from lignocellulosic substrates (Wang et al 2018) and the ability to grow at high temperatures (Fu et al 2019). Recently, mainly using transcriptome analysis, we determined that young genes specific to K. marxianus are enriched in the response to stresses such as high temperature, low pH and high osmolarity (Doughty et al 2020).…”

Section: Introductionmentioning

confidence: 99%

Development of a Ribosome Profiling Protocol to Study Translation in the yeast Kluyveromyces marxianus

Fenton

Kiniry

Yordanova

et al. 2022

Preprint

View full text Add to dashboard Cite

Kluyveromyces marxianus is an interesting and important yeast because of particular traits like thermotolerance and rapid growth, and applications in food and industrial biotechnology. Knowing how K. marxianus responds and adapts to changing environments is important to achieve a full understanding of the its biology and to develop bioprocesses. For this, a full suite of omics tools to measure and compare global patterns of gene expression and protein synthesis is needed. Whereas transcriptome analysis by RNA-Seq quantifies mRNA abundance, ribosome profiling allows codon-resolution of translation on a genome-wide scale by deep sequencing of ribosome locations on mRNAs and is emerging as a valuable tool to study translation control of gene expression. We report here the development of a ribosome profiling method for K. marxianus and we make the procedure available as a step by step protocol. To aid in the analysis and sharing of ribosome profiling data, we also added the K. marxianus genome as well as transcriptome and ribosome profiling data to the publicly accessible GWIPS-Viz and Trips-Viz browsers. Users are able to upload custom ribosome profiling and RNA-Seq data to both browsers, therefore allowing easy analysis and sharing of data. As many studies only focus on the use of RNA-Seq to study K. marxianus in different environments, the availability of ribosome profiling is a powerful addition to the K. marxianus toolbox.

show abstract

Ethanol production from xylose is highly increased by the Kluyveromyces marxianus mutant 17694-DH1

Cited by 14 publications

References 29 publications

Alleviation of catabolite repression in Kluyveromyces marxianus: the thermotolerant SBK1 mutant simultaneously coferments glucose and xylose

Alleviation of catabolite repression in Kluyveromyces marxianus: the thermotolerant SBK1 mutant simultaneously coferments glucose and xylose

Kluyveromyces marxianus developing ethanol tolerance during adaptive evolution with significant improvements of multiple pathways

Development of a Ribosome Profiling Protocol to Study Translation in the yeast Kluyveromyces marxianus

Contact Info

Product

Resources

About