Differential RNA-seq, Multi-Network Analysis and Metabolic Regulation Analysis of Kluyveromyces marxianus Reveals a Compartmentalised Response to Xylose

Schabort, Du Toit W. P.; Letebele, Precious K.; Steyn, Laurinda; Kilian, S. G.; Preez, J. C. du

doi:10.1371/journal.pone.0156242

Cited by 31 publications

(40 citation statements)

References 44 publications

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“…Growth rate was significantly slower with xylose (0.25 hr −1 ) and lactose (0.52 hr −1 ) relative to glucose (0.66 hr −1 ), and less ethanol, glycerol, and organic acid byproducts were formed in selective xylose cultures of CBS712Δ URA3 (Figure S3). Increased byproduct formation in glucose relative to xylose has been previously observed, as well as reduced transcription of amino acid synthesis genes in xylose (Schabort, Letebele, Steyn, Kilian, & Du Preez, ) which may contribute to slower growth rates.…”

Section: Resultsmentioning

confidence: 88%

“…The minimal byproduct formation observed in xylose may contribute to the higher levels of TAL produced relative to glucose; however, these byproducts do not account for the significant difference in titers. Transcription analyses for K. marxianus report greater than 100‐fold higher transcription of fatty acid and lipid catabolism genes in xylose relative to glucose as well as increased conversion of ethanol via the TCA cycle (Lertwattanasakul et al, ; Schabort et al, ); these may contribute to higher acetyl‐CoA for conversion to TAL. While the full metabolism of K. marxianus in glucose and xylose is still not fully understood, important factors such as lipid degradation and accumulation, byproduct formation, cofactor balances, sugar transport and growth rate as well as putative phosphoketolase activity (Evans & Ratledge, ) may contribute to the improved growth and titers from xylose.…”

Section: Resultsmentioning

confidence: 99%

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Synthesis of polyketides from low cost substrates by the thermotolerant yeast Kluyveromyces marxianus

McTaggart

Bever

Bassett

et al. 2019

Biotech & Bioengineering

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Kluyveromyces marxianus is a promising nonconventional yeast for biobased chemical production due to its rapid growth rate, high TCA cycle flux, and tolerance to low pH and high temperature. Unlike Saccharomyces cerevisiae, K. marxianus grows on low-cost substrates to cell densities that equal or surpass densities in glucose, which can be beneficial for utilization of lignocellulosic biomass (xylose), biofuel production waste (glycerol), and whey (lactose). We have evaluated K. marxianus for the synthesis of polyketides, using triacetic acid lactone (TAL) as the product. The 2-pyrone synthase (2-PS) was expressed on a CEN/ARS plasmid in three different strains, and the effects of temperature, carbon source, and cultivation strategy on TAL levels were determined.The highest titer was obtained in defined 1% xylose medium at 37°C, with substantial titers at 41 and 43°C. The introduction of a high-stability 2-PS mutant and a promoter substitution increased titer four-fold. 2-PS expression from a multi-copy pKD1-based plasmid improved TAL titers a further five-fold. Combining the best plasmid, promoter, and strain resulted in a TAL titer of 1.24 g/L and a yield of 0.0295 mol TAL/mol carbon for this otherwise unengineered strain in 3 ml tube culture. This is an excellent titer and yield (on xylose) before metabolic engineering or fed-batch culture relative to other hosts (on glucose), and demonstrates the promise of this rapidly growing and thermotolerant yeast species for polyketide production.

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Section: Resultsmentioning

confidence: 88%

Section: Resultsmentioning

confidence: 99%

Synthesis of polyketides from low cost substrates by the thermotolerant yeast Kluyveromyces marxianus

McTaggart

Bever

Bassett

et al. 2019

Biotech & Bioengineering

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show abstract

“…In the case of unconventional thermotolerant yeast K. marxianus, however, there were only several reports of transcriptome analysis on yeast responses from different carbon source including glucose, inulin and xylose, or the tolerance to high temperature or ethanol stress. 18,69,70,71 To the best of our knowledge, genome-wide transcription analysis under the multiple inhibitors stress condition with elevated temperature (42 C) has not been reported for this yeast.…”

Section: Discussionmentioning

confidence: 99%

Transcriptomic analysis of thermotolerant yeastKluyveromyces marxianusin multiple inhibitors tolerance

Wang

Yang

et al. 2018

RSC Adv.

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During pretreatment of lignocellulosic biomass, toxic compounds were released and inhibited the growth and fermentation of microorganisms. Here the global transcriptional response of K. marxianus to multiple inhibitors including acetic acid, phenols, furfural and HMF, at 42 C, was studied, via RNA-seq technology.Genes involved in the glycolysis pathway, fatty acid metabolism, ergosterol metabolism and vitamin B6 and B1 metabolic process were enriched in the down-regulated gene set, while genes involved in TCA cycle, respiratory chain, ROS detoxification and transporter coding genes were enriched in the up-regulated gene set in response to the multiple inhibitors stress. Further real time-PCR results with three single inhibitor stress conditions showed that different transporters responded quite differently to different inhibitor stress. Coenzyme assay results showed that the level of NAD + was increased and both NADH/ NAD + and NADPH/NADP + ratio decreased. Furthermore, genes involved with transcription factors related to carbohydrate metabolism, sulfur amino acids metabolism, lipid metabolism or those directly involved in the transcriptional process were significantly regulated. Though belonging to different GO categories or KEGG pathway, many differentially expressed genes were enriched in maintaining the redox balance, NAD(P) + /NAD(P)H homeostasis or NAD + synthesis, energy production, and iron transportation or metabolism. These results suggest that engineering these aspects represents a possible strategy to develop more robust strains for industrial fermentation from cellulosic biomass.

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“…Genomic and transcriptomic studies have started to shed light on K. marxianus, and a growing number of genome sequences of K. marxianus strains are now available. Those include KCTC 17555 [34], DMB1 [95], CCT 7735 [96], NBRC1777 [97], DMKU 3-1042 [35], B0399 [98], UFS-Y2791 [99], and other nine strains: L01, L02, L03, L04, L05, CBS397, NBRC0272, NBRC0288, and NBRC0617 [100].…”

Section: Complete Genome Sequence Of Thermotolerant Yeast K Marxianumentioning

confidence: 99%

Potential of Thermotolerant Ethanologenic Yeasts Isolated from ASEAN Countries and Their Application in High- Temperature Fermentation

Kosaka¹,

Lertwattanasakul²,

NadchanokRodrussamee³

et al. 2019

Fuel Ethanol Production From Sugarcane

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Thermotolerant ethanologenic yeasts receive attention as alternative bio-ethanol producers to traditionally used yeast, Saccharomyces cerevisiae. Their utilization is expected to provide several benefits for bio-ethanol production due to their characteristics and robustness. They have been isolated from a wide variety of environments in a number of ASEAN countries: Thailand, Vietnam, Laos, and Indonesia. One of these yeasts, Kluyveromyces marxianus has been investigated regarding characteristics. Some strains efficiently utilize xylose, which is a main component of the 2nd generation biomass. In addition, the genetic basis of K. marxianus has been revealed by genomic sequencing and is exploited for further improvement of the strains by thermal adaptation or gene engineering techniques. Moreover, the glucose repression of K. marxianus and its mechanisms has been investigated. Results suggest that K. marxianus is an alternative to S. cerevisiae in next-generation bio-ethanol production industry. Indeed, we have succeeded to apply K. marxianus for bio-ethanol production in a newly developed process, which combines high-temperature fermentation with simultaneous fermentation and distillation under low pressure. This chapter aims to provide valuable information on thermotolerant ethanologenic yeasts and their application, which may direct the economic bioproduction of ethanol and other useful materials in the future.Recently, thermotolerant microorganisms were found among mesophiles with optimum growth temperatures that are 5-10°C higher than those of the typical mesophilic strains Fuel Ethanol Production from Sugarcane 122 Potential of Thermotolerant Ethanologenic Yeasts Isolated from ASEAN Countries… http://dx.doi.org/10.5772/intechopen.79144 123Recently, there have been several reports on ethanol production at high temperatures using P. kudriavzevii (formerly known as I. orientalis). Several P. kudriavzevii strains were reported to grow and produce high levels of ethanol at high temperatures. The strain DMKU 3-ET15 was isolated from traditional fermented pork sausage in Thailand by an enrichment technique in a medium supplemented with 4% ethanol at 40°C. The strain produced 78.6 g/L ethanol from 180 g/L glucose at 40°C [20]. The strain KVMP10 that was isolated from soil located beneath apple trees for ethanol production from orange peel achieved 54 g/L ethanol at 42°C [48]. Strain RZ8-1 that was recently isolated from various samples collected from plant orchards in Thailand produced 33.8 g/L ethanol from 160 g/L glucose at 40°C [49].

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Differential RNA-seq, Multi-Network Analysis and Metabolic Regulation Analysis of Kluyveromyces marxianus Reveals a Compartmentalised Response to Xylose

Cited by 31 publications

References 44 publications

Synthesis of polyketides from low cost substrates by the thermotolerant yeast Kluyveromyces marxianus

Synthesis of polyketides from low cost substrates by the thermotolerant yeast Kluyveromyces marxianus

Transcriptomic analysis of thermotolerant yeastKluyveromyces marxianusin multiple inhibitors tolerance

Potential of Thermotolerant Ethanologenic Yeasts Isolated from ASEAN Countries and Their Application in High- Temperature Fermentation

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