scholarly journals Protein acetylation regulates xylose metabolism during adaptation of Saccharomyces cerevisiae

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Yong-Shui Tan ◽  
Li Wang ◽  
Ying-Ying Wang ◽  
Qi-En He ◽  
Zhi-Hua Liu ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Xylose Reductase ◽  
Xylose Isomerase ◽  
Xylose Utilization ◽  
Xylose Metabolism ◽  
Xylose Consumption ◽  
Synthetic Complete ◽  
Before And After ◽  
Genes Encoding ◽  
Fuels And Chemicals

Abstract Background As the second most abundant polysaccharide in nature, hemicellulose can be degraded to xylose as the feedstock for bioconversion to fuels and chemicals. To enhance xylose conversion, the engineered Saccharomyces cerevisiae with xylose metabolic pathway is usually adapted with xylose as the carbon source in the laboratory. However, the mechanism under the adaptation phenomena of the engineered strain is still unclear. Results In this study, xylose-utilizing S. cerevisiae was constructed and used for the adaptation study. It was found that xylose consumption rate increased 1.24-fold in the second incubation of the yYST12 strain in synthetic complete-xylose medium compared with the first incubation. The study figured out that it was observed at the single-cell level that the stagnation time for xylose utilization was reduced after adaptation with xylose medium in the microfluidic device. Such transient memory of xylose metabolism after adaptation with xylose medium, named “xylose consumption memory”, was observed in the strains with both xylose isomerase pathway and xylose reductase and xylitol dehydrogenase pathways. In further, the proteomic acetylation of the strains before and after adaptation was investigated, and it was revealed that H4K5 was one of the most differential acetylation sites related to xylose consumption memory of engineered S. cerevisiae. We tested 8 genes encoding acetylase or deacetylase, and it was found that the knockout of the GCN5 and HPA2 encoding acetylases enhanced the xylose consumption memory. Conclusions The behavior of xylose consumption memory in engineered S. cerevisiae can be successfully induced with xylose in the adaptation. H4K5Ac and two genes of GCN5 and HPA2 are related to xylose consumption memory of engineered S. cerevisiae during adaptation. This study provides valuable insights into the xylose adaptation of engineered S. cerevisiae.

scholarly journals Protein acetylation regulates xylose metabolism during adaptation of Saccharomyces cerevisiae

2021 ◽  
Author(s):  
Yong-Shui Tan ◽  
Li Wang ◽  
Ying-Ying Wang ◽  
Qi-En He ◽  
Zhen Zhu ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Fold Increase ◽  
The State ◽  
Confirmatory Test ◽  
Xylose Utilization ◽  
Protein Acetylation ◽  
Xylose Metabolism ◽  
Promising Alternative ◽  
Xylose Consumption ◽  
Synthetic Complete

Abstract Background Lignocellulosic biomass upgrading has become a promising alternative route to produce transportation fuels in response to energy security and environmental concerns. As the second most abundant polysaccharide in nature, hemicellulose mainly containing xylose is an important carbon source that can be used for the bioconversion to fuels and chemicals. However, the adaptation phenomena could appear and influence the bioconversion performance of xylose when Saccharomyces cerevisiae strain was transferred from the glucose to the xylose environment. Therefore, it is crucial to elucidate the mechanism of this adaptation phenomena, which can guide the strategy exploration to improve the efficiency of xylose utilization. Results In this study, xylose-utilizing strains had been constructed to effectively consume xylose. It is found that the second incubation of yYST218 strain in synthetic complete-xylose medium resulted in a 1.24-fold increase in xylose consumption ability as compared with the first incubation in synthetic complete-xylose medium. The results clearly showed that growing S. cerevisiae again in synthetic complete-xylose medium can significantly reduce the stagnation time and thus achieved a faster growth rate, by comparing the growth status of the strain in synthetic complete-xylose medium for the first and second time at the single-cell level through Microfluidic technology. Although these xylose-utilizing strains possessed different xylose metabolism pathways, they exhibited the “transient memory” phenomenon of xylose metabolism after changing the culture environment to synthetic complete-xylose medium, which named ‘xylose consumption memory (XCM)’ of S. cerevisiae in this study. According to the identification of protein acetylation, partial least squares analysis and the confirmatory test had verified that H4K5Ac affected the state of “XCM” in S. cerevisiae. Knockout of the acetylase-encoding genes GCN5 and HPA2 enhanced the “XCM” of the strain. Protein acetylation analysis suggested that xylose induced perturbation in S. cerevisiae stimulated the rapid adaptation of strains to xylose environment by regulating the level of acetylation. Conclusions All these results indicated protein acetylation modification is an important aspect that protein acetylation regulated the state of “XCM” in S. cerevisiae and thus determine the environmental adaptation of S. cerevisiae. Systematically exploiting the regulation approach of protein acetylation in S. cerevisiae could provide valuable insights into the adaptation phenomena of microorganisms in complex industrial environments.

scholarly journals Directed Evolution of Xylose Isomerase for Improved Xylose Catabolism and Fermentation in the Yeast Saccharomyces cerevisiae

2012 ◽  
Vol 78 (16) ◽  
pp. 5708-5716 ◽  
Author(s):  
Sun-Mi Lee ◽  
Taylor Jellison ◽  
Hal S. Alper
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ethanol Production ◽  
Directed Evolution ◽  
Ethanol Yield ◽  
Xylose Isomerase ◽  
Mutant Enzyme ◽  
Xylose Utilization ◽  
Xylose Consumption ◽  
Content Type ◽  
Consumption Rates

ABSTRACTThe heterologous expression of a highly functional xylose isomerase pathway inSaccharomyces cerevisiaewould have significant advantages for ethanol yield, since the pathway bypasses cofactor requirements found in the traditionally used oxidoreductase pathways. However, nearly all reported xylose isomerase-based pathways inS. cerevisiaesuffer from poor ethanol productivity, low xylose consumption rates, and poor cell growth compared with an oxidoreductase pathway and, additionally, often require adaptive strain evolution. Here, we report on the directed evolution of thePiromycessp. xylose isomerase (encoded byxylA) for use in yeast. After three rounds of mutagenesis and growth-based screening, we isolated a variant containing six mutations (E15D, E114G, E129D, T142S, A177T, and V433I) that exhibited a 77% increase in enzymatic activity. When expressed in a minimally engineered yeast host containing agre3knockout andtal1andXKS1overexpression, the strain expressing this mutant enzyme improved its aerobic growth rate by 61-fold and both ethanol production and xylose consumption rates by nearly 8-fold. Moreover, the mutant enzyme enabled ethanol production by these yeasts under oxygen-limited fermentation conditions, unlike the wild-type enzyme. Under microaerobic conditions, the ethanol production rates of the strain expressing the mutant xylose isomerase were considerably higher than previously reported values for yeast harboring a xylose isomerase pathway and were also comparable to those of the strains harboring an oxidoreductase pathway. Consequently, this study shows the potential to evolve a xylose isomerase pathway for more efficient xylose utilization.

scholarly journals Directed evolution and secretory expression of xylose isomerase for improved utilisation of xylose in Saccharomyces cerevisiae

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Jung-Hoon Bae ◽  
Mi-Jin Kim ◽  
Bong Hyun Sung ◽  
Yong-Su Jin ◽  
Jung-Hoon Sohn
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Directed Evolution ◽  
Lignocellulosic Biomass ◽  
Xylose Fermentation ◽  
Xylose Isomerase ◽  
Carbon Substrate ◽  
Yeast Fermentation ◽  
Xylose Utilization ◽  
Secretory Expression ◽  
Xylose Consumption

Abstract Background Xylose contained in lignocellulosic biomass is an attractive carbon substrate for economically viable conversion to bioethanol. Extensive research has been conducted on xylose fermentation using recombinant Saccharomyces cerevisiae expressing xylose isomerase (XI) and xylose reductase/xylitol dehydrogenase (XR/XDH) pathways along with the introduction of a xylose transporter and amplification of the downstream pathway. However, the low utilization of xylose in the presence of glucose, due to the varying preference for cellular uptake, is a lingering challenge. Studies so far have mainly focused on xylose utilization inside the cells, but there have been little trials on the conversion of xylose to xylulose by cell before uptake. We hypothesized that the extracellular conversion of xylose to xylulose before uptake would facilitate better utilization of xylose even in the presence of glucose. To verify this, XI from Piromyces sp. was engineered and hyper-secreted in S. cerevisiae for the extracellular conversion of xylose to xylulose. Results The optimal pH of XI was lowered from 7.0 to 5.0 by directed evolution to ensure its high activity under the acidic conditions used for yeast fermentation, and hyper-secretion of an engineered XI-76 mutant (E56A and I252M) was accomplished by employing target protein-specific translational fusion partners. The purified XI-76 showed twofold higher activity than that of the wild type at pH 5. The secretory expression of XI-76 in the previously developed xylose utilizing yeast strain, SR8 increased xylose consumption and ethanol production by approximately 7–20% and 15–20% in xylose fermentation and glucose and xylose co-fermentation, respectively. Conclusions Isomerisation of xylose to xylulose before uptake using extracellular XI was found to be effective in xylose fermentation or glucose/xylose co-fermentation. This suggested that glucose competed less with xylulose than with xylose for uptake by the cell. Consequently, the engineered XI secretion system constructed in this study can pave the way for simultaneous utilization of C5/C6 sugars from the sustainable lignocellulosic biomass.

scholarly journals Development and characterization of efficient xylose utilization strains of Zymomonas mobilis

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Jiyun Lou ◽  
Jingwen Wang ◽  
Yongfu Yang ◽  
Qing Yang ◽  
Runxia LI ◽  
...  
Keyword(s):  
Recombinant Strain ◽  
Consumption Rate ◽  
Xylose Isomerase ◽  
Xylose Utilization ◽  
Rna Seq ◽  
Strain Development ◽  
Genome Resequencing ◽  
Xylose Consumption ◽  
Genes Encoding ◽  
Lignocellulosic Biofuels

Abstract Background Efficient use of glucose and xylose is a key for the economic production of lignocellulosic biofuels and biochemicals, and different recombinant strains have been constructed for xylose utilization including those using Zymomonas mobilis as the host. However, the xylose utilization efficiency still needs to be improved. In this work, the strategy of combining metabolic engineering and adaptive laboratory evolution (ALE) was employed to develop recombinant Z. mobilis strains that can utilize xylose efficiently at high concentrations, and NGS-based genome resequencing and RNA-Seq transcriptomics were performed for strains evolved after serial transfers in different media to understand the impact of xylose and differences among strains with different xylose-utilization capabilities at molecular level. Results Heterologous genes encoding xylose isomerase and xylulokinase were evaluated, which were then introduced into xylose-utilizing strain Z. mobilis 8b to enhance its capacity of xylose utilization. The results demonstrated that the effect of three xylose isomerases on xylose utilization was different, and the increase of copy number of xylose metabolism genes can improve xylose utilization. Among various recombinant strains constructed, the xylose utilization capacity of the recombinant strain 8b-RsXI-xylB was the best, which was further improved through continuous adaption with 38 transfers over 100 days in 50 g/L xylose media. The fermentation performances of the parental strain 8b, the evolved 8b-S38 strain with the best xylose utilization capability, and the intermediate strain 8b-S8 in different media were compared, and the results showed that only 8b-S38 could completely consume xylose at 50 g/L and 100 g/L concentrations. In addition, the xylose consumption rate of 8b-S38 was faster than that of 8b at different xylose concentrations from 50 to 150 g/L, and the ethanol yield increased by 16 ~ 40%, respectively. The results of the mixed-sugar fermentation also demonstrated that 8b-S38 had a higher xylose consumption rate than 8b, and its maximum ethanol productivity was 1.2 ~ 1.4 times higher than that of 8b and 8b-S8. Whole-genome resequencing identified three common genetic changes in 8b-S38 compared with 8b and 8b-S8. RNA-Seq study demonstrated that the expression levels of genes encoding chaperone proteins, ATP-dependent proteases, phage shock proteins, ribosomal proteins, flagellar operons, and transcriptional regulators were significantly increased in xylose media in 8b-S38. The up-regulated expression of these genes may therefore contribute to the efficient xylose utilization of 8b-S38 by maintaining the normal cell metabolism and growth, repairing cellular damages, and rebalancing cellular energy to help cells resist the stressful environment. Conclusions This study provides gene candidates to improve xylose utilization, and the result of expressing an extra copy of xylose isomerase and xylulokinase improved xylose utilization also provides a direction for efficient xylose-utilization strain development in other microorganisms. In addition, this study demonstrated the necessity to combine metabolic engineering and ALE for industrial strain development. The recombinant strain 8b-S38 can efficiently metabolize xylose for ethanol fermentation at high xylose concentrations as well as in mixed sugars of glucose and xylose, which could be further developed as the microbial biocatalyst for the production of lignocellulosic biofuels and biochemicals.

scholarly journals Simulating Extracellular Glucose Signals Enhances Xylose Metabolism in Recombinant Saccharomyces cerevisiae

2020 ◽  
Vol 8 (1) ◽  
pp. 100 ◽  
Author(s):  
Meiling Wu ◽  
Hongxing Li ◽  
Shan Wei ◽  
Hongyu Wu ◽  
Xianwei Wu ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Metabolic Networks ◽  
Glucose Sensing ◽  
Biofuel Production ◽  
Xylose Utilization ◽  
High Concentration ◽  
Xylose Metabolism ◽  
Camp Phosphodiesterase ◽  
Xylose Consumption ◽  
Recombinant Saccharomyces Cerevisiae

Efficient utilization of both glucose and xylose from lignocellulosic biomass would be economically beneficial for biofuel production. Recombinant Saccharomyces cerevisiae strains with essential genes and metabolic networks for xylose metabolism can ferment xylose; however, the efficiency of xylose fermentation is much lower than that of glucose, the preferred carbon source of yeast. Implications from our previous work suggest that activation of the glucose sensing system may benefit xylose metabolism. Here, we show that deleting cAMP phosphodiesterase genes PDE1 and PDE2 increased PKA activity of strains, and consequently, increased xylose utilization. Compared to the wild type strain, the specific xylose consumption rate (rxylose) of the pde1Δ pde2Δ mutant strains increased by 50%; the specific ethanol-producing rate (rethanol) of the strain increased by 70%. We also show that HXT1 and HXT2 transcription levels slightly increased when xylose was present. We also show that HXT1 and HXT2 transcription levels slightly increased when xylose was present. Deletion of either RGT2 or SNF3 reduced expression of HXT1 in strains cultured in 1 g L−1 xylose, which suggests that xylose can bind both Snf3 and Rgt2 and slightly alter their conformations. Deletion of SNF3 significantly weakened the expression of HXT2 in the yeast cultured in 40 g L−1 xylose, while deletion of RGT2 did not weaken expression of HXT2, suggesting that S. cerevisiae mainly depends on Snf3 to sense a high concentration of xylose (40 g L−1). Finally, we show that deletion of Rgt1, increased rxylose by 24% from that of the control. Our findings indicate how S. cerevisiae may respond to xylose and this study provides novel targets for further engineering of xylose-fermenting strains.

scholarly journals Molecular Analysis of a Saccharomyces cerevisiae Mutant with Improved Ability To Utilize Xylose Shows Enhanced Expression of Proteins Involved in Transport, Initial Xylose Metabolism, and the Pentose Phosphate Pathway

2003 ◽  
Vol 69 (2) ◽  
pp. 740-746 ◽  
Author(s):  
C. Fredrik Wahlbom ◽  
Ricardo R. Cordero Otero ◽  
Willem H. van Zyl ◽  
Bärbel Hahn-Hägerdal ◽  
Leif J. Jönsson
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Pentose Phosphate Pathway ◽  
Specific Activity ◽  
Xylose Reductase ◽  
Pentose Phosphate ◽  
Percentage Error ◽  
Xylose Utilization ◽  
Xylose Metabolism ◽  
Physiological Characterization ◽  
Cell Extracts

ABSTRACT Differences between the recombinant xylose-utilizing Saccharomyces cerevisiae strain TMB 3399 and the mutant strain TMB 3400, derived from TMB 3399 and displaying improved ability to utilize xylose, were investigated by using genome-wide expression analysis, physiological characterization, and biochemical assays. Samples for analysis were withdrawn from chemostat cultures. The characteristics of S. cerevisiae TMB 3399 and TMB 3400 grown on glucose and on a mixture of glucose and xylose, as well as of S. cerevisiae TMB 3400 grown on only xylose, were investigated. The strains were cultivated under chemostat conditions at a dilution rate of 0.1 h−1, with feeds consisting of a defined mineral medium supplemented with 10 g of glucose liter−1, 10 g of glucose plus 10 g of xylose liter−1 or, for S. cerevisiae TMB 3400, 20 g of xylose liter−1. S. cerevisiae TMB 3400 consumed 31% more xylose of a feed containing both glucose and xylose than S. cerevisiae TMB 3399. The biomass yields for S. cerevisiae TMB 3400 were 0.46 g of biomass g of consumed carbohydrate−1 on glucose and 0.43 g of biomass g of consumed carbohydrate−1 on xylose. A Ks value of 33 mM for xylose was obtained for S. cerevisiae TMB 3400. In general, the percentage error was <20% between duplicate microarray experiments originating from independent fermentation experiments. Microarray analysis showed higher expression in S. cerevisiae TMB 3400 than in S. cerevisiae TMB 3399 for (i) HXT5, encoding a hexose transporter; (ii) XKS1, encoding xylulokinase, an enzyme involved in one of the initial steps of xylose utilization; and (iii) SOL3, GND1, TAL1, and TKL1, encoding enzymes in the pentose phosphate pathway. In addition, the transcriptional regulators encoded by YCR020C, YBR083W, and YPR199C were expressed differently in the two strains. Xylose utilization was, however, not affected in strains in which YCR020C was overexpressed or deleted. The higher expression of XKS1 in S. cerevisiae TMB 3400 than in TMB 3399 correlated with higher specific xylulokinase activity in the cell extracts. The specific activity of xylose reductase and xylitol dehydrogenase was also higher for S. cerevisiae TMB 3400 than for TMB 3399, both on glucose and on the mixture of glucose and xylose.

scholarly journals Saccharomyces cerevisiae Engineered for Xylose Metabolism Exhibits a Respiratory Response

2004 ◽  
Vol 70 (11) ◽  
pp. 6816-6825 ◽  
Author(s):  
Yong-Su Jin ◽  
Jose M. Laplaza ◽  
Thomas W. Jeffries
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Tricarboxylic Acid Cycle ◽  
Pentose Phosphate ◽  
Oxygen Limitation ◽  
Sugar Uptake ◽  
Xylose Utilization ◽  
Xylose Metabolism ◽  
Expression Of Genes ◽  
Genes Encoding ◽  
Native Strains

ABSTRACT Native strains of Saccharomyces cerevisiae do not assimilate xylose. S. cerevisiae engineered for d-xylose utilization through the heterologous expression of genes for aldose reductase (XYL1), xylitol dehydrogenase (XYL2), and d-xylulokinase (XYL3 or XKS1) produce only limited amounts of ethanol in xylose medium. In recombinant S. cerevisiae expressing XYL1, XYL2, and XYL3, mRNA transcript levels for glycolytic, fermentative, and pentose phosphate enzymes did not change significantly on glucose or xylose under aeration or oxygen limitation. However, expression of genes encoding the tricarboxylic acid cycle, respiration enzymes (HXK1, ADH2, COX13, NDI1, and NDE1), and regulatory proteins (HAP4 and MTH1) increased significantly when cells were cultivated on xylose, and the genes for respiration were even more elevated under oxygen limitation. These results suggest that recombinant S. cerevisiae does not recognize xylose as a fermentable carbon source and that respiratory proteins are induced in response to cytosolic redox imbalance; however, lower sugar uptake and growth rates on xylose might also induce transcripts for respiration. A petite respiration-deficient mutant (ρ�) of the engineered strain produced more ethanol and accumulated less xylitol from xylose. It formed characteristic colonies on glucose, but it did not grow on xylose. These results are consistent with the higher respiratory activity of recombinant S. cerevisiae when growing on xylose and with its inability to grow on xylose under anaerobic conditions.

scholarly journals Deletion ofFPS1, Encoding Aquaglyceroporin Fps1p, Improves Xylose Fermentation by Engineered Saccharomyces cerevisiae

2013 ◽  
Vol 79 (10) ◽  
pp. 3193-3201 ◽  
Author(s):  
Na Wei ◽  
Haiqing Xu ◽  
Soo Rin Kim ◽  
Yong-Su Jin
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Xylose Fermentation ◽  
Plant Biomass ◽  
Ethanol Yield ◽  
Xylose Reductase ◽  
Enzymatic Reactions ◽  
Xylose Metabolism ◽  
Xylose Consumption ◽  
Content Type ◽  
Ethanol Yields

ABSTRACTAccumulation of xylitol in xylose fermentation with engineeredSaccharomyces cerevisiaepresents a major problem that hampers economically feasible production of biofuels from cellulosic plant biomass. In particular, substantial production of xylitol due to unbalanced redox cofactor usage by xylose reductase (XR) and xylitol dehydrogenase (XDH) leads to low yields of ethanol. While previous research focused on manipulating intracellular enzymatic reactions to improve xylose metabolism, this study demonstrated a new strategy to reduce xylitol formation and increase carbon flux toward target products by controlling the process of xylitol secretion. Using xylitol-producingS. cerevisiaestrains expressing XR only, we determined the role of aquaglyceroporin Fps1p in xylitol export by characterizing extracellular and intracellular xylitol. In addition, whenFPS1was deleted in a poorly xylose-fermenting strain with unbalanced XR and XDH activities, the xylitol yield was decreased by 71% and the ethanol yield was substantially increased by nearly four times. Experiments with our optimized xylose-fermenting strain also showed thatFPS1deletion reduced xylitol production by 21% to 30% and increased ethanol yields by 3% to 10% under various fermentation conditions. Deletion ofFPS1decreased the xylose consumption rate under anaerobic conditions, but the effect was not significant in fermentation at high cell density. Deletion ofFPS1resulted in higher intracellular xylitol concentrations but did not significantly change the intracellular NAD+/NADH ratio in xylose-fermenting strains. The results demonstrate that Fps1p is involved in xylitol export inS. cerevisiaeand present a new gene deletion target,FPS1, and a mechanism different from those previously reported to engineer yeast for improved xylose fermentation.

scholarly journals Deletion of the GRE3 Aldose Reductase Gene and Its Influence on Xylose Metabolism in Recombinant Strains of Saccharomyces cerevisiae Expressing thexylA and XKS1 Genes

2001 ◽  
Vol 67 (12) ◽  
pp. 5668-5674 ◽  
Author(s):  
K. L. Träff ◽  
R. R. Otero Cordero ◽  
W. H. van Zyl ◽  
B. Hahn-Hägerdal
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Aldose Reductase ◽  
Thermus Thermophilus ◽  
Xylose Isomerase ◽  
Xylose Utilization ◽  
Xylose Metabolism ◽  
Gene Encoding ◽  
Recombinant Strains ◽  
Ethanol Formation ◽  
Bacterial Enzyme

ABSTRACT Saccharomyces cerevisiae ferments hexoses efficiently but is unable to ferment xylose. When the bacterial enzyme xylose isomerase (XI) from Thermus thermophilus was produced in S. cerevisiae, xylose utilization and ethanol formation were demonstrated. In addition, xylitol and acetate were formed. An unspecific aldose reductase (AR) capable of reducing xylose to xylitol has been identified inS. cerevisiae. The GRE3gene, encoding the AR enzyme, was deleted in S.cerevisiae CEN.PK2-1C, yielding YUSM1009a. XI fromT. thermophilus was produced, and endogenous xylulokinase from S.cerevisiae was overproduced in S.cerevisiae CEN.PK2-1C and YUSM1009a. In recombinant strains from which the GRE3 gene was deleted, xylitol formation decreased twofold. Deletion of the GRE3 gene combined with expression of the xylA gene fromT. thermophilus on a replicative plasmid generated recombinant xylose utilizing S.cerevisiae strain TMB3102, which produced ethanol from xylose with a yield of 0.28 mmol of C from ethanol/mmol of C from xylose. None of the recombinant strains grew on xylose.

scholarly journals Recombinant Diploid Saccharomyces cerevisiae Strain Development for Rapid Glucose and Xylose Co-Fermentation

Fermentation ◽  
2018 ◽  
Vol 4 (3) ◽  
pp. 59 ◽  
Author(s):  
Tingting Liu ◽  
Shuangcheng Huang ◽  
Anli Geng
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Alcoholic Fermentation ◽  
Ethanol Yield ◽  
Xylose Isomerase ◽  
Cost Effective ◽  
Xylose Utilization ◽  
Yeast Strains ◽  
Multiple Copies ◽  
Biomass Sugars ◽  
Sugar Conversion

Cost-effective production of cellulosic ethanol requires robust microorganisms for rapid co-fermentation of glucose and xylose. This study aims to develop a recombinant diploid xylose-fermenting Saccharomyces cerevisiae strain for efficient conversion of lignocellulosic biomass sugars to ethanol. Episomal plasmids harboring codon-optimized Piromyces sp. E2 xylose isomerase (PirXylA) and Orpinomyces sp. ukk1 xylose (OrpXylA) genes were constructed and transformed into S. cerevisiae. The strain harboring plasmids with tandem PirXylA was favorable for xylose utilization when xylose was used as the sole carbon source, while the strain harboring plasmids with tandem OrpXylA was beneficial for glucose and xylose cofermentation. PirXylA and OrpXylA genes were also individually integrated into the genome of yeast strains in multiple copies. Such integration was beneficial for xylose alcoholic fermentation. The respiration-deficient strain carrying episomal or integrated OrpXylA genes exhibited the best performance for glucose and xylose co-fermentation. This was partly attributed to the high expression levels and activities of xylose isomerase. Mating a respiration-efficient strain carrying the integrated PirXylA gene with a respiration-deficient strain harboring integrated OrpXylA generated a diploid recombinant xylose-fermenting yeast strain STXQ with enhanced cell growth and xylose fermentation. Co-fermentation of 162 g L−1 glucose and 95 g L−1 xylose generated 120.6 g L−1 ethanol in 23 h, with sugar conversion higher than 99%, ethanol yield of 0.47 g g−1, and ethanol productivity of 5.26 g L−1·h−1.

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