Dataset Information

Xylitol production from non-detoxified and non-sterile lignocellulosic hydrolysate using low-cost industrial media components.

ABSTRACT: Immobilized Candida tropicalis cells in freeze dried calcium alginate beads were used for production of xylitol from lignocellulosic waste like corn cob hydrolysate without any detoxification and sterilization of media. Media components for xylitol fermentation were screened by statistical methods. Urea, KH2PO4 and initial pH were identified as significant variables by Plackett-Burman (PB) design. Significant medium components were optimized by response surface methodology (RSM). Predicted xylitol yield by RSM model and experimental yield was 0.87 and 0.79 g/g, respectively. Optimized conditions (urea 1.5 g/L, KH2PO4 1.9 g/L, xylose 55 g/L, pH 6.7) enhanced xylitol yield by 32% and xylose consumption by twofold over those of basal media. In addition, the immobilized cells were reused five times at shake flask level with optimized medium without affecting the xylitol productivity and yield. Xylitol production was successfully scaled up to 7.5 L stirred tank reactor using optimized media. Thus, the optimized condition with non-detoxified pentose hydrolysate from corn cob lignocellulosic waste with minimal nutrients without any sterilization opens up the scope for commercialization of the process.

SUBMITTER: Yewale T

PROVIDER: S-EPMC5428108 | biostudies-literature | 2017 May

REPOSITORIES: biostudies-literature

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Xylitol production from non-detoxified and non-sterile lignocellulosic hydrolysate using low-cost industrial media components.

Yewale Tatyaso T Panchwagh Shruti S Sawale Shaileshkumar S Jain Rishi R Dhamole Pradip B PB

3 Biotech 20170427 1

Immobilized Candida tropicalis cells in freeze dried calcium alginate beads were used for production of xylitol from lignocellulosic waste like corn cob hydrolysate without any detoxification and sterilization of media. Media components for xylitol fermentation were screened by statistical methods. Urea, KH<sub>2</sub>PO<sub>4</sub> and initial pH were identified as significant variables by Plackett-Burman (PB) design. Significant medium components were optimized by response surface methodology ...[more]

PMID: 28452022

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Project description:BackgroundIntegrated management of hemicellulosic fraction and its economical transformation to value-added products is the key driver towards sustainable lignocellulosic biorefineries. In this aspect, microbial cell factories are harnessed for the sustainable production of commercially viable biochemicals by valorising C5 and C6 sugars generated from agro-industrial waste. However, in the terrestrial ecosystem, microbial systems can efficiently consume glucose. On the contrary, pentose sugars are less preferred carbon source as most of the microbes lack metabolic pathway for their utilization. The effective utilization of both pentose and hexose sugars is key for economical biorefinery.ResultsBioprospecting the food waste and selective enrichment on xylose-rich medium led to screening and isolation of yeast which was phylogenetically identified as Pichia fermentans. The newly isolated xylose assimilating yeast was explored for xylitol production. The wild type strain robustly grew on xylose and produced xylitol with >?40% conversion yield. Chemical mutagenesis of isolated yeast with ethyl methanesulphonate (EMS) yielded seven mutants. The mutant obtained after 15 min EMS exposure, exhibited best xylose bioconversion efficiency. This mutant under shake flask conditions produced maximum xylitol titer and yield of 34.0 g/L and 0.68 g/g, respectively. However, under the same conditions, the control wild type strain accumulated 27.0 g/L xylitol with a conversion yield of 0.45 g/g. Improved performance of the mutant was attributed to 34.6% activity enhancement in xylose reductase with simultaneous reduction of xylitol dehydrogenase activity by 22.9%. Later, the culture medium was optimized using statistical design and validated at shake flask and bioreactor level. Bioreactor studies affirmed the competence of the mutant for xylitol accumulation. The xylitol titer and yield obtained with pure xylose were 98.9 g/L and 0.67 g/g, respectively. In comparison, xylitol produced using non-detoxified xylose rich pre-hydrolysate from sugarcane bagasse was 79.0 g/L with an overall yield of 0.54 g/g.ConclusionThis study demonstrates the potential of newly isolated P. fermentans in successfully valorising the hemicellulosic fraction for the sustainable xylitol production.

Project description:UnlabelledA major obstacle to sustainable lignocellulosic biofuel production is microbe inhibition by the combinatorial stresses in pretreated plant hydrolysate. Chemical biomass pretreatment releases a suite of toxins that interact with other stressors, including high osmolarity and temperature, which together can have poorly understood synergistic effects on cells. Improving tolerance in industrial strains has been hindered, in part because the mechanisms of tolerance reported in the literature often fail to recapitulate in other strain backgrounds. Here, we explored and then exploited variations in stress tolerance, toxin-induced transcriptomic responses, and fitness effects of gene overexpression in different Saccharomyces cerevisiae (yeast) strains to identify genes and processes linked to tolerance of hydrolysate stressors. Using six different S. cerevisiae strains that together maximized phenotypic and genetic diversity, first we explored transcriptomic differences between resistant and sensitive strains to identify common and strain-specific responses. This comparative analysis implicated primary cellular targets of hydrolysate toxins, secondary effects of defective defense strategies, and mechanisms of tolerance. Dissecting the responses to individual hydrolysate components across strains pointed to synergistic interactions between osmolarity, pH, hydrolysate toxins, and nutrient composition. By characterizing the effects of high-copy gene overexpression in three different strains, we revealed the breadth of the background-specific effects of gene fitness contributions in synthetic hydrolysate. Our approach identified new genes for engineering improved stress tolerance in diverse strains while illuminating the effects of genetic background on molecular mechanisms.ImportanceRecent studies on natural variation within Saccharomyces cerevisiae have uncovered substantial phenotypic diversity. Here, we took advantage of this diversity, using it as a tool to infer the effects of combinatorial stress found in lignocellulosic hydrolysate. By comparing sensitive and tolerant strains, we implicated primary cellular targets of hydrolysate toxins and elucidated the physiological states of cells when exposed to this stress. We also explored the strain-specific effects of gene overexpression to further identify strain-specific responses to hydrolysate stresses and to identify genes that improve hydrolysate tolerance independent of strain background. This study underscores the importance of studying multiple strains to understand the effects of hydrolysate stress and provides a method to find genes that improve tolerance across strain backgrounds.

Dataset Information

Xylitol production from non-detoxified and non-sterile lignocellulosic hydrolysate using low-cost industrial media components.

Publications

Xylitol production from non-detoxified and non-sterile lignocellulosic hydrolysate using low-cost industrial media components.

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