Xiaoyu Tang

Transcriptome profiling of Zymomonas mobilis under furfural stress

Furfural from lignocellulosic hydrolysates is the prevalent inhibitor to microorganisms during cellulosic ethanol production, but the molecular mechanisms of tolerance to this inhibitor in Zymomonas mobilis are still unclear. In this study, genome-wide transcriptional responses to furfural were investigated in Z. mobilis using microarray analysis. We found that 433 genes were differentially expressed in response to furfural. Furfural up- or down-regulated genes related to cell wall/membrane biogenesis, metabolism, and transcription. However, furfural has a subtle negative effect on Entner–Doudoroff pathway mRNAs. Our results revealed that furfural had effects on multiple aspects of cellular metabolism at the transcriptional level and that membrane might play important roles in response to furfural. This research has provided insights into the molecular response to furfural in Z. mobilis, and it will be helpful to construct more furfural-resistant strains for cellulosic ethanol production.

Engineered hydrochar composites for phosphorus removal/recovery: Lanthanum doped hydrochar prepared by hydrothermal carbonization of lanthanum pretreated rice straw.

Engineered hydrochar composites (EHC) were synthesized by hydrothermal carbonization (HTC) of lanthanum pretreated rice straw. The as-prepared composite with about 30% lanthanum content showed greater P removal potential than La(OH)3, indicating the synergistic effect of hydrochar and lanthanum in P removal. The adsorption results showed that EHC showed great P adsorption capacities (>50mgPg(-1)) in the pH range of 2.5-10.5, and the presence of competing anions had little negative effects on P adsorption on EHC. The equilibrium time for P adsorption on EHC was considerably reduced under acid condition (12h) compared to alkaline condition (48h). The maximum adsorption capacity was 61.57mgPg(-1) according to Langmuir isotherms. These results suggested that EHC was highly effective in P adsorption in a wide range of pH and the presence of competing anions, thus EHC could be a promising adsorbent for phosphorus removal/recovery from wastewater.

Bamboo: a new source of carbohydrate for biorefinery.

Bamboo is perennial woody grass, which distributed widely in the world and belonged to the Gramineae family and Bambuseae subfamily. It may be consider as a candidate lignocellulosic substrate for bio-ethanol production for its environmental benefits and higher annual biomass yield. The conversion of bamboo into bio-ethanol, bio-methane, natural food, flavonoids, and functional xylo-oligosaccharides production were reviewed in this paper. Future prospects for research include pretreatment, enzymatic hydrolysis and fermentation will also be performed to improve the whole process of ethanol production more economical. And revealing the molecular regulation mechanism of the fast growth of bamboo will provide chance for improving bamboo or other energy plants biomass yield through genetic engineering.

Adaptive laboratory evolution of ethanologenic Zymomonas mobilis strain tolerant to furfural and acetic acid inhibitors.

Furfural and acetic acid from lignocellulosic hydrolysates are the prevalent inhibitors to Zymomonas mobilis during cellulosic ethanol production. Developing a strain tolerant to furfural or acetic acid inhibitors is difficul by using rational engineering strategies due to poor understanding of their underlying molecular mechanisms. In this study, strategy of adaptive laboratory evolution (ALE) was used for development of a furfural and acetic acid-tolerant strain. After three round evolution, four evolved mutants (ZMA7-2, ZMA7-3, ZMF3-2, and ZMF3-3) that showed higher growth capacity were successfully obtained via ALE method. Based on the results of profiling of cell growth, glucose utilization, ethanol yield, and activity of key enzymes, two desired strains, ZMA7-2 and ZMF3-3, were achieved, which showed higher tolerance under 7xa0g/l acetic acid and 3xa0g/l furfural stress condition. Especially, it is the first report of Z. mobilis strain that could tolerate higher furfural. The best strain, Z. mobilis ZMF3-3, has showed 94.84xa0% theoretical ethanol yield under 3-g/l furfural stress condition, and the theoretical ethanol yield of ZM4 is only 9.89xa0%. Our study also demonstrated that ALE method might also be used as a powerful metabolic engineering tool for metabolic engineering in Z. mobilis. Furthermore, the two best strains could be used as novel host for further metabolic engineering in cellulosic ethanol or future biorefinery. Importantly, the two strains may also be used as novel-tolerant model organisms for the genetic mechanism on the “omics” level, which will provide some useful information for inverse metabolic engineering.

Effects of high ammonium level on biomass accumulation of common duckweed Lemna minor L.

Growing common duckweed Lemna minor L. in diluted livestock wastewater is an alternative option for pollutants removal and consequently the accumulated duckweed biomass can be used for bioenergy production. However, the biomass accumulation can be inhibited by high level of ammonium (NH4+) in non-diluted livestock wastewater and the mechanism of ammonium inhibition is not fully understood. In this study, the effect of high concentration of NH4+ on L. minor biomass accumulation was investigated using NH4+ as sole source of nitrogen (N). NH4+-induced toxicity symptoms were observed when L. minor was exposed to high concentrations of ammonium nitrogen (NH4+-N) after a 7-day cultivation. L. minor exposed to the NH4+-N concentration of 840xa0mgxa0l−1 exhibited reduced relative growth rate, contents of carbon (C) and photosynthetic pigments, and C/N ratio. Ammonium irons were inhibitory to the synthesis of photosynthetic pigments and caused C/N imbalance in L. minor. These symptoms could further cause premature senescence of the fronds, and restrain their reproduction, growth and biomass accumulation. L. minor could grow at NH4+-N concentrations of 7–84xa0mgxa0l−1 and the optimal NH4+-N concentration was 28xa0mgxa0l−1.

Transcriptomic and physiological analysis of common duckweed Lemna minor responses to NH4+ toxicity

BackgroundPlants can suffer ammonium (NH4+) toxicity, particularly when NH4+ is supplied as the sole nitrogen source. However, our knowledge about the underlying mechanisms of NH4+ toxicity is still largely unknown. Lemna minor, a model duckweed species, can grow well in high NH4+ environment but to some extent can also suffer toxic effects. The transcriptomic and physiological analysis of L. minor responding to high NH4+ may provide us some interesting and useful information not only in toxic processes, but also in tolerance mechanisms.ResultsThe L. minor cultured in the Hoagland solution were used as the control (NC), and in two NH4+ concentrations (NH4+ was the sole nitrogen source), 84xa0mg/L (A84) and 840xa0mg/L (A840) were used as stress treatments. The NH4+ toxicity could inhibit the growth of L. minor. Reactive oxygen species (ROS) and cell death were studied using stained fronds under toxic levels of NH4+. The malondialdehyde content and the activities of superoxide dismutase and peroxidase increased from NC to A840, rather than catalase and ascorbate peroxidase. A total of 6.62G nucleotides were generated from the three distinct libraries. A total of 14,207 differentially expressed genes (DEGs) among 70,728 unigenes were obtained. All the DEGs could be clustered into 7 profiles. Most DEGs were down-regulated under NH4+ toxicity. The genes required for lignin biosynthesis in phenylpropanoid biosynthesis pathway were up-regulated. ROS oxidative-related genes and programmed cell death (PCD)-related genes were also analyzed and indicated oxidative damage and PCD occurring under NH4+ toxicity.ConclusionsThe first large transcriptome study in L. minor responses to NH4+ toxicity was reported in this work. NH4+ toxicity could induce ROS accumulation that causes oxidative damage and thus induce cell death in L. minor. The antioxidant enzyme system was activated under NH4+ toxicity for ROS scavenging. The phenylpropanoid pathway was stimulated under NH4+ toxicity. The increased lignin biosynthesis might play an important role in NH4+ toxicity resistance.

Predicting the impacts of climate change on the potential distribution of major native non-food bioenergy plants in China.

Planting non-food bioenergy crops on marginal lands is an alternative bioenergy development solution in China. Native non-food bioenergy plants are also considered to be a wise choice to reduce the threat of invasive plants. In this study, the impacts of climate change (a consensus of IPCC scenarios A2a for 2080) on the potential distribution of nine non-food bioenergy plants native to China (viz., Pistacia chinensis, Cornus wilsoniana, Xanthoceras sorbifolia, Vernicia fordii, Sapium sebiferum, Miscanthus sinensis, M. floridulus, M. sacchariflorus and Arundo donax) were analyzed using a MaxEnt species distribution model. The suitable habitats of the nine non-food plants were distributed in the regions east of the Mongolian Plateau and the Tibetan Plateau, where the arable land is primarily used for food production. Thus, the large-scale cultivation of those plants for energy production will have to rely on the marginal lands. The variables of “precipitation of the warmest quarter” and “annual mean temperature” were the most important bioclimatic variables for most of the nine plants according to the MaxEnt modeling results. Global warming in coming decades may result in a decrease in the extent of suitable habitat in the tropics but will have little effect on the total distribution area of each plant. The results indicated that it will be possible to grow these plants on marginal lands within these areas in the future. This work should be beneficial for the domestication and cultivation of those bioenergy plants and should facilitate land-use planning for bioenergy crops in China.

Carbon and energy fixation of great duckweed Spirodela polyrhiza growing in swine wastewater

The ability to fix carbon and energy in swine wastewater of duckweeds was investigated using Spirodela polyrhiza as the model species. Cultures of S. polyrhiza were grown in dilutions of both original swine wastewater (OSW) and anaerobic digestion effluent (ADE) based on total ammonia nitrogen (TAN). Results showed that elevated concentrations of TAN caused decreased growth, carbon fixation, and energy production rates, particularly just after the first rise in two types of swine wastewater. Also, OSW was more suitable for S. polyrhiza cultivation than ADE. Maximum carbon and energy fixation were achieved at OSW-TAN concentrations of 12.08 and 13.07xa0mgxa0L−1, respectively. Photosynthetic activity of S. polyrhiza could be inhibited by both nutrient stress (in high-concentration wastewater) and nutrient limitation (in low-concentration wastewater), affecting its growth and ability for carbon-energy fixation.

Construction of a novel secretion expression system guided by native signal peptide of PhoD in Zymomonas mobilis

In the current study, three native signal peptides (SPs) from PhoC, PhoD, and ZMO0331were investigated and compared to construct novel secretion expression systems in Zymomonas mobilis. The secretion expression of target protein, α-amylase from Bacillus amyloliquefaciens (BAA), guided by PhoD’s SP resulted in more hydrolysis of starch than that by the other two SPs. Extracellular and intracellular α-amylase activities of the strain containing PhoD’s SP were also higher than the other two strains containing PhoC or ZMO0331’s SP. In addition, the evidence by alcohol dehydrogenase activity assay further confirmed that the starch hydrolysis was resulted from the secretion expression of BAA rather than the breakage of cells. Our results indicated that the SP of PhoD is able to serve as a promising candidate to assist secretion expression of heterogeneous genes in Z. mobilis. This will contribute to development of engineered Z. mobilis strains converting starch into ethanol.

Additional file 2: Figure S1. of Transcriptomic and physiological analysis of common duckweed Lemna minor responses to NH4 + toxicity

Sequencing saturation analysis (A) and distribution of gene coverage (B) in each library. (DOCX 78 kb)