Establishment of Kluyveromyces marxianus as a Microbial Cell Factory for Lignocellulosic Processes: Production of High Value Furan Derivatives

Abstract

The establishment of lignocellulosic biorefineries is dependent on microorganisms being able to cope with the stressful conditions resulting from the release of inhibitory compounds during biomass processing. The yeast Kluyveromyces marxianus has been explored as an alternative microbial factory due to its thermotolerance and ability to natively metabolize xylose. The lignocellulose-derived inhibitors furfural and 5-hydroxymethylfurfural (HMF) are considered promising building-block platforms that can be converted into a wide variety of high-value derivatives. Here, several K. marxianus strains, isolated from cocoa fermentation, were evaluated for xylose consumption and tolerance towards acetic acid, furfural, and HMF. The potential of this yeast to reduce furfural and HMF at high inhibitory loads was disclosed and characterized. Our results associated HMF reduction with NADPH while furfural-reducing activity was higher with NADH. In addition, furans’ inhibitory effect was higher when combined with xylose consumption. The furan derivatives produced by K. marxianus in different conditions were identified. Furthermore, one selected isolate was efficiently used as a whole-cell biocatalyst to convert furfural and HMF into their derivatives, furfuryl alcohol and 2,5-bis(hydroxymethyl)furan (BHMF), with high yields and productivities. These results validate K. marxianus as a promising microbial platform in lignocellulosic biorefineries.

Transcriptional and translational profiling in yeast reveals the use of diverse genome decoding mechanisms to generate functionally distinct proteoforms

“…More recently, its capacity for rapid growth, thermotolerance and other relevant traits has garnered much attention in the biotechnology sector (Fonseca et al ., 2008; Lane and Morrissey, 2010; Morrissey et al ., 2015; Varela et al ., 2017; Karim, Gerliani and Aïder, 2020). This has led to substantial progress in the development of gene engineering and synthetic biology tools (Nambu-Nishida et al ., 2017; Cernak et al ., 2018; Rajkumar et al ., 2019; Rajkumar and Morrissey, 2022), meaning that it is now relatively straightforward to reprogramme strains as cell factories for industrial biotechnology applications (Liu and Nielsen, 2019; Rajkumar and Morrissey, 2020; Baptista, Cunha and Domingues, 2021; Patra et al ., 2021). Other developments at the physiological level are improving the potential for large-scale fermentation of K. marxianus under industrial conditions (Dekker et al ., 2021).…”

Dissecting an ancient stress resistance trait syndrome in the compost yeastKluyveromyces marxianus

“…And though K. marxianus can be isolated from rotting fruit 40,41 , it may not have specialized narrowly to such substrates: a neighboring clade that includes Saccharomyces cerevisiae produces more ethanol than do any of the Kluyveromyces species 6 , suggesting a weaker drive in the latter toward this putative mechanism to kill off microbial competitors in high-sugar environments. Perhaps most compelling among microbial ecology findings is the prevalence of K. marxianus in decomposing plant material beside fruit, including leaf litter and pine needles 42 , cow dung 19 and industrially processed cocoa beans 43 , agave 44 , and sugar cane 16,45 . If the ancestral niche for K. marxianus included plant substrates of this kind, the yeast could have specialized in rapid metabolism of free sugars, leaving the breakdown of lignocellulosic plant cell wall components to other fungal saprotrophs.…”

Whole Cell Biocatalysis of 5-Hydroxymethylfurfural for Sustainable Biorefineries

“…The bacterium Burkholderia contaminans NJPI-15 reached the highest BHMF titer of 656 mM in a fed-batch with 94% yield [29]. More recently, the highest BHMF yield of 99.65% was reported using a Kluyveromyces marxianus yeast strain isolated from cocoa fermentation, which reached an BHMF titer of 55.3 mM in a batch process [30].…”