Potential Carbon Source Research Articles

Loving the poison: the methylcitrate cycle and bacterial pathogenesis.

Propionate is an abundant catabolite in nature and represents a rich potential source of carbon for the organisms that can utilize it. However, propionate and propionate-derived catabolites are also toxic to cells, so propionate catabolism can alternatively be viewed as a detoxification mechanism. In this review, we summarize recent progress made in understanding how prokaryotes catabolize propionic acid, how these pathways are regulated and how they might be exploited to develop novel antibacterial interventions.

The Effects of Different External Carbon Sources on Nitrous Oxide Emissions during Denitrification in Biological Nutrient Removal Processes

The aim of this study was to investigate the effects of two different external carbon sources (acetate and ethanol) on the nitrous oxide (N2O) emissions during denitrification in biological nutrient removal processes. Results showed that external carbon source significantly influenced N2O emissions during the denitrification process. When acetate served as the external carbon source, 0.49 mg N/L and 0.85 mg N/L of N2O was produced during the denitrificaiton processes in anoxic and anaerobic/anoxic experiments, giving a ratio of N2O-N production to TN removal of 2.37% and 4.96%, respectively. Compared with acetate, the amount of N2O production is negligible when ethanol used as external carbon addition. This suggested that ethanol is a potential alternative external carbon source for acetate from the point of view of N2O emissions.

Structural insights into marine carbohydrate degradation by family GH16 κ-carrageenases

Carrageenans are sulfated α-1,3-β-1,4-galactans found in the cell wall of some red algae that are practically valuable for their gelation and biomimetic properties but also serve as a potential carbon source for marine bacteria. Carbohydrate degradation has been studied extensively for terrestrial plant/bacterial systems, but sulfation is not present in these cases, meaning the marine enzymes used to degrade carrageenans must possess unique features to recognize these modifications. To gain insights into these features, we have focused on κ-carrageenases from two distant bacterial phyla, which belong to glycoside hydrolase family 16 and cleave the β-1,4 linkage of κ-carrageenan. We have solved the crystal structure of the catalytic module of ZgCgkA from Zobellia galactanivorans at 1.66 Å resolution and compared it with the only other structure available, that of PcCgkA from Pseudoalteromonas carrageenovora 9T (ATCC 43555T). We also describe the first substrate complex in the inactivated mutant form of PcCgkA at 1.7 Å resolution. The structural and biochemical comparison of these enzymes suggests key determinants that underlie the functional properties of this subfamily. In particular, we identified several arginine residues that interact with the polyanionic substrate, and confirmed the functional relevance of these amino acids using a targeted mutagenesis strategy. These results give new insight into the diversity of the κ-carrageenase subfamily. The phylogenetic analyses show the presence of several distinct clades of enzymes that relate to differences in modes of action or subtle differences within the same substrate specificity, matching the hybrid character of the κ-carrageenan polymer.

Low-Temperature and Rapid Growth of Large Single-Crystalline Graphene with Ethane.

Future applications of graphene rely highly on the production of large-area high-quality graphene, especially large single-crystalline graphene, due to the reduction of defects caused by grain boundaries. However, current large single-crystalline graphene growing methodologies are suffering from low growth rate and as a result, industrial graphene production is always confronted by high energy consumption, which is primarily caused by high growth temperature and long growth time. Herein, a new growth condition achieved via ethane being the carbon feedstock to achieve low-temperature yet rapid growth of large single-crystalline graphene is reported. Ethane condition gives a growth rate about four times faster than methane, achieving about 420 µm min-1 for the growth of sub-centimeter graphene single crystals at temperature about 1000 °C. In addition, the temperature threshold to obtain graphene using ethane can be reduced to 750 °C, lower than the general growth temperature threshold (about 1000 °C) with methane on copper foil. Meanwhile ethane always keeps higher graphene growth rate than methane under the same growth temperature. This study demonstrates that ethane is indeed a potential carbon source for efficient growth of large single-crystalline graphene, thus paves the way for graphene in high-end electronical and optoelectronical applications.

Analysis of graphene-like activated carbon derived from rice straw for application in supercapacitor

Activated carbons with large surface area, abundant microporosity and low cost are the most commonly used electrode materials for energy storage devices. However, activated carbons are conventionally made from fossil precursors, such as coal and petroleum, which are limited resources and easily aggregate large block in high temperature carbonization processes. In this novel work, we examined the use of rice straw as a potential alternative carbon source precursor for the production of graphene-like active carbon. A very slack activated carbon with ultra-thin two-dimensional (2D) layer structure was prepared by our proposed approach in this work, which includes a pre-treatment process and potassium hydroxide activation at high temperatures. The obtained active carbon derived from rice straw exhibited a capacitance of 255F/g at 0.5A/g, excellent rate capability, and long cycling capability (98% after 10,000 cycles).

Simultaneous valorization and detoxification of the hemicellulose rich liquor from the organosolv fractionation

Fractionation of lignocellulosic biomass with solvents (organosolv process) generates a hemicellulose-rich liquor with a high content of phenolics which is particularly toxic. This work addresses the utilization of this stream as a potential carbon source for the production of ligninolytic enzymes (LE). Among six basidiomycetes species, Irpex lacteus and Ganoderma lucidum presented the highest activities of manganese peroxidase (646 ± 122 U L−1) and laccase (1497 ± 161 U L−1), respectively, growing on a medium composed mainly of cellulose fibers, lignin and hemicellulose. The influence of each lignocellulosic fraction on the LE production mechanisms was studied in more detail. The high concentration of phenolic compounds in the hemicellulose-rich stream acted as inducer of LE production, with levels even greater than those of xylose. Acute toxicity tests on Vibrio fischeri revealed a substantial reduction of the toxicity after the fungal treatment (by ca. 12–21 times). The proposed valorization and detoxification of this currently non-exploited and abundant by-product stream is a promising strategy to enhance the industrial feasibility of the organosolv fractionation process.

Metagenomic analysis reveals potential interactions in an artificial coculture

Disentangling the interactions between cyanobacteria and associated bacterial community is important for understanding the mechanisms that mediate the formation of cyanobacterial blooms in freshwater ecosystems. Despite the fact that a metagenomic approach enables researchers to profile the structure of microbial communities associated with cyanobacteria, reconstructing genome sequences for all members remains inefficient, due to the inherent enormous microbial diversity. Here, we have established a stable coculture system under high salinity, originally from a mixture of an axenic cyanobacterium Synechococcus sp. PCC 7002 and a non-axenic bloom-forming cyanobacterium Microcystis colony. Metagenomic analysis showed that the coculture consists of S. sp. PCC 7002 and two heterotrophic bacteria, designated as Pseudomonas stutzeri TAIHU and Mesorhizobium sp. TAIHU, respectively. And near-complete genome sequences of both bacteria were reconstructed from the metagenomic dataset with an average completeness of 99.8%. Genome-wide pathway analysis revealed that M. sp. TAIHU carried all the genes involved in the de novo biosynthesis of cobalamin, which is required by S. sp. PCC 7002 for growth. To cope with the high salinity in the coculture, experimental evidence demonstrated that S. sp. PCC 7002 would synthesize the compatible solutes including sucrose and glucosylglycerol, which are supposed to be exploited by both heterotrophic bacteria as potential carbon and/or nitrogen sources. Furthermore, the genes encoding for the biosynthesis of the ectoine, another common osmolyte are found exclusively in P. stutzeri TAIHU, while the genes responsible for the catabolism of ectoine and its derives are present only in M. sp. TAIHU. These genomic evidence indicates beneficial interaction between three members in the coculture. Establishment of the coculture system with relative simplicity provides a useful model system for investigating the interspecies interactions, and genome sequences of both bacteria associated with Microcystis bloom described here will facilitate the researcher to elucidate the role of these heterotrophic bacteria in the formation and maintenance of cyanobacterial bloom in freshwater ecosystem.

Synthesizing value-added products from methane by a new Methylomonas.

Methane and methanol are potential carbon sources of industrial micro-organisms in addition to crop-derived bio-carbon sources. Methanotrophs that can utilize these simple, stable and large amounts chemicals are expected to be developed into 'cell factories' for the production of specific chemicals. In this study, a methanotroph that can synthesize lycopene, C30 carotenoid and exopolysaccharides (EPS) with relative better performances from C1 substrates was isolated, and its performances were evaluated. The isolated strain was identified as Methylomonas sp. ZR1 based on 16S rRNA sequence analysis. Its maximum specific growth rate achieved 0·200h-1 under flask culture conditions, and 0·386h-1 in bubble column reactors. ZR1 was able to utilize 35gl-1 of methanol and even exhibited slight growth in the presence of 40gl-1 of methanol. Furthermore, ZR1 was proved to synthesize lycopene (C40 carotenoids) besides the C30 carotenoids through HPLC-DAD and HPLC-MS/MS analysis methods. Its carotenoid extracts exhibited excellent antioxidative activities measured by the ABTS+ method. Plenty of polysaccharides were also synthesized by ZR1, the components of the polysaccharides were identified as glucose, mannose and galactose with a proportion of 1:2:1 by GC-MS, and its yield achieved 0·13gg-1 cell dry weight. The isolated strain has great potential for the production of value-added bioproducts from C1 compounds because of its excellent C1 substrate utilizing abilities and its abilities to naturally synthesize lycopene, C30 carotenoids and EPS. In this study, we isolated a fast-growing methanotroph, its C1 carbon substrate utilizing ability is excellent in comparison with reported methanotrophs. Furthermore, besides polysaccharides and C30 carotenoids which were commonly synthesized by methanotrophs, our findings suggested that C40 lycopene could also be naturally synthesized from methane by methanotrophs.

Gene Duplication in Pseudomonas aeruginosa Improves Growth on Adenosine.

The laboratory strain of Pseudomonas aeruginosa, PAO1, activates genes for catabolism of adenosine using quorum sensing (QS). However, this strain is not well-adapted for growth on adenosine, with doubling times greater than 40 h. We previously showed that when PAO1 is grown on adenosine and casein, variants emerge that grow rapidly on adenosine. To understand the mechanism by which this adaptation occurs, we performed whole-genome sequencing of five isolates evolved for rapid growth on adenosine. All five genomes had a gene duplication-amplification (GDA) event covering several genes, including the quorum-regulated nucleoside hydrolase gene, nuh, and PA0148, encoding an adenine deaminase. In addition, two of the growth variants also exhibited a nuh promoter mutation. We recapitulated the rapid growth phenotype with a plasmid containing six genes common to all the GDA events. We also showed that nuh and PA0148, the two genes at either end of the common GDA, were sufficient to confer rapid growth on adenosine. Additionally, we demonstrated that the variant nuh promoter increased basal expression of nuh but maintained its QS regulation. Therefore, GDA in P. aeruginosa confers the ability to grow efficiently on adenosine while maintaining QS regulation of nucleoside catabolism.IMPORTANCEPseudomonas aeruginosa thrives in many habitats and is an opportunistic pathogen of humans. In these diverse environments, P. aeruginosa must adapt to use myriad potential carbon sources. P. aeruginosa PAO1 cannot grow efficiently on nucleosides, including adenosine; however, it can acquire this ability through genetic adaptation. We show that the mechanism of adaptation is by amplification of a specific region of the genome and that the amplification preserves the regulation of the adenosine catabolic pathway by quorum sensing. These results demonstrate an underexplored mechanism of adaptation by P. aeruginosa, with implications for phenotypes such as development of antibiotic resistance.

Tracing carbon transfer and assimilation by invertebrates and fish across a tropical mangrove ecosystem using stable isotopes

AbstractMangroves are highly productive ecosystems that exhibit a diverse range of habitats, including tidal creeks and flats, forest gaps and interior forest with varying understory light intensity, tidal dynamics, geomorphological settings, and overall biological production. Within mangrove ecosystems, invertebrates and fish feed on heterogeneous food sources, the occurrence of which is unevenly distributed across the system. This provides a basis for testing models of carbon transfer across mangrove ecosystems. We hypothesized that the carbon transfer and assimilation by fish and invertebrates will vary across the different mangrove habitats and that such variations can be predicted by their stable isotope compositions. We analysed δ13C and δ15N signatures of consumers and their potential organic carbon sources across a tropical mangrove ecosystem in Vietnam. The δ13C values of crabs and snails significantly decreased from the tidal flat to interior forest, indicating that variations in carbon transfer and assimilation occurred at small scales <30 m. Reduced variation in δ13C of suspension‐feeding bivalves suggested that tidal water was a vector for large‐scale transport of carbon across the mangrove ecosystem. An analysis of co‐variance using habitat as a fixed factor and feeding habit and movement capacity of consumers as co‐variates indicated that habitat and feeding types were major features that affected the δ13C values of invertebrates and fish. The findings demonstrate that carbon transfer and assimilation across mangrove ecosystems occur as a diverse combination of small (<30 m) and large (>30 m) scale processes.

Atmospheric pCO2 reconstructed across five early Eocene global warming events

Multiple short-lived global warming events, known as hyperthermals, occurred during the early Eocene (56–52 Ma). Five of these events – the Paleocene–Eocene Thermal Maximum (PETM or ETM1), H1 (or ETM2), H2, I1, and I2 – are marked by a carbon isotope excursion (CIE) within both marine and terrestrial sediments. The magnitude of CIE, which is a function of the amount and isotopic composition of carbon added to the ocean–atmosphere system, varies significantly between marine versus terrestrial substrates. Here we use the increase in carbon isotope fractionation by C3 land plants in response to increased pCO2 to reconcile this difference and reconstruct a range of background pCO2 and peak pCO2 for each CIE, provided two potential carbon sources: methane hydrate destabilization and permafrost-thawing/organic matter oxidation. Although the uncertainty on each pCO2 estimate using this approach is low (e.g., median uncertainty = +23%/−18%), this work highlights the potential for significant systematic bias in the pCO2 estimate resulting from sampling resolution, substrate type, diagenesis, and environmental change. Careful consideration of each of these factors is required especially when applying this approach to a single marine–terrestrial CIE pair. Given these limitations, we provide an upper estimate for background early Eocene pCO2 of 463 +248/−131 ppmv (methane hydrate scenario) to 806 +127/−104 ppmv (permafrost-thawing/organic matter oxidation scenario). These results, which represent the first pCO2 proxy estimates directly tied to the Eocene hyperthermals, demonstrate that early Eocene warmth was supported by background pCO2 less than ∼3.5× preindustrial levels and that pCO2>1000 ppmv may have occurred only briefly, during hyperthermal events.

Modelling past, present and future peatland carbon accumulation across the pan-Arctic region

Abstract. Most northern peatlands developed during the Holocene, sequestering large amounts of carbon in terrestrial ecosystems. However, recent syntheses have highlighted the gaps in our understanding of peatland carbon accumulation. Assessments of the long-term carbon accumulation rate and possible warming-driven changes in these accumulation rates can therefore benefit from process-based modelling studies. We employed an individual-based dynamic global ecosystem model with dynamic peatland and permafrost functionalities and patch-based vegetation dynamics to quantify long-term carbon accumulation rates and to assess the effects of historical and projected climate change on peatland carbon balances across the pan-Arctic region. Our results are broadly consistent with published regional and global carbon accumulation estimates. A majority of modelled peatland sites in Scandinavia, Europe, Russia and central and eastern Canada change from carbon sinks through the Holocene to potential carbon sources in the coming century. In contrast, the carbon sink capacity of modelled sites in Siberia, far eastern Russia, Alaska and western and northern Canada was predicted to increase in the coming century. The greatest changes were evident in eastern Siberia, north-western Canada and in Alaska, where peat production hampered by permafrost and low productivity due the cold climate in these regions in the past was simulated to increase greatly due to warming, a wetter climate and higher CO2 levels by the year 2100. In contrast, our model predicts that sites that are expected to experience reduced precipitation rates and are currently permafrost free will lose more carbon in the future.

Evaluation of Sludge Palm Oil as Feedstock and Development of Efficient Method for its Utilization to Produce Polyhydroxyalkanoate

Sludge palm oil (SPO), a difficult-to-be-used solid byproduct of the palm oil milling industry, was evaluated as potential carbon source for poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx)] production using recombinant Cupriavidus necator Re2058/pCB113. The biosynthesis of polyhydroxyalkanoate (PHA) was conducted using single stage shake flask growth to study different parameters affecting the bacterial growth. Fed-batch fermentations were conducted using SPO to increase the PHA productivity and to reach high density culture which is deemed necessary for large scale production. Initial shake flask studies showed that SPO can be utilized by the bacteria for growth and PHA accumulation. However, the yield of SPO conversion into cell biomass and PHA was low due to the difficulty in using the solid oil in liquid medium. With the aid of surfactant and mixing strategy, SPO which consisted mainly of free fatty acids was successfully emulsified in the mineral medium and was used for cell growth and PHA accumulation whereby 10 g/L of SPO supplied as emulsion solution produced 9.7 g/L of CDW containing 74 wt% P(3HB-co-22 mol% 3HHx). The high yield of biomass obtained indicates that SPO is an excellent feedstock for this strain. Fed-batch fermentation was conducted to increase the yield and productivity whereby a biomass productivity of 1.9 g/L/h and PHA productivity of 1.1 g/L/h were achieved. These results suggest that SPO, an inexpensive waste material, can be used to produce PHA in large scale for commercialization purpose with reduced production cost.

Fermentable sugars production by enzymatic processing of agave leaf juice

AbstractThe Mexican mezcal industry annually processes approximately 2.92 × 105 t of mezcal agave, generating roughly 1.46 × 105 t of agave leaves per year, which represents a potential carbon source of at least 8170 t via enzymatic processing of agave leaf juice. This carbon source is considered an attractive alternative to produce biofuels and/or chemical products since it is produced and used without adversely affecting the environment. The aim of this investigation was to determine the effect of temperature, pH, enzyme concentration, and bioreaction time on the enzymatic hydrolysis of agave leaf juice enriched in fructan to maximize the fermentable sugars production from three varieties of mezcal agave, using a low‐cost commercial brand of hydrolase. This process generated a sugar‐enriched juice of 80.07–136.12 g/L of reducing sugars. A Box‐Behnken experimental design and a mathematical surface response analysis of the hydrolysis were used for process optimization.

Decomposition characteristics of three different kinds of aquatic macrophytes and their potential application as carbon resource in constructed wetland

Decomposition of aquatic macrophytes usually generates significant influence on aquatic environment. Study on the aquatic macrophytes decomposition may help reusing the aquatic macrophytes litters, as well as controlling the water pollution caused by the decomposition process. This study verified that the decomposition processes of three different kinds of aquatic macrophytes (water hyacinth, hydrilla and cattail) could exert significant influences on water quality of the receiving water, including the change extent of pH, dissolved oxygen (DO), the contents of carbon, nitrogen and phosphorus, etc. The influence of decomposition on water quality and the concentrations of the released chemical materials both followed the order of water hyacinth > hydrilla > cattail. Greater influence was obtained with higher dosage of plant litter addition. The influence also varied with sediment addition. Moreover, nitrogen released from the decomposition of water hyacinth and hydrilla were mainly NH3-N and organic nitrogen while those from cattail litter included organic nitrogen and NO3−-N. After the decomposition, the average carbon to nitrogen ratio (C/N) in the receiving water was about 2.6 (water hyacinth), 5.3 (hydrilla) and 20.3 (cattail). Therefore, cattail litter might be a potential plant carbon source for denitrification in ecological system of a constructed wetland.

Food web foundation and seasonal variation of trophic structure based on the stable isotopic technique in the marine ranching of Haizhou Bay, China

Stable isotope technique was applied to investigate the small and medium-size consumers including not only fish, shrimps, crabs, cephalopods, snails and bivalves, but also the specimens of their potential carbon sources, which were collected in the marine ranching area of Haizhou Bay during the spring of 2015. IsoSource model was adopted to calculate the contribution of potential carbon sources to the consumers. Biospeciemens collected in the summer of 2014 were compared with those in the spring of 2015, aiming to analyze the seasonal variation in the trophic structure of food web. Six quantitative community indices were calculated based on the isotope values of the organisms in Haizhou Bay. The results showed that the δ13C values of consumers ranged from -18.9‰ to -17.1‰, and the δ13C values of three potential food sources ranged from -18.1‰ to -23.4‰ in the spring of 2015. The calculation results of IsoSource model further demonstrated that the phytoplankton was the most important carbon source for consumers (80.8%), followed by sediment organic matter (10.8%) and particulate organic matter (8.4%). There was a significant difference in the δ13C values between the biospecimens in the summer of 2014 and those in the spring of 2015, while no significant difference was observed in δ15N values. Trophic structure of community-wide differed remarkably in different seasons indicated by the six quantitative community indices. The δ13C range, total area, mean nearest neighbor distance and standard deviation of nearest neighbor distance of the community were much higher in 2014 than those in 2015, but the δ15N range and mean distance to centroid showed no significant change. The trophic structure redundancy was lower in the summer of 2014 compared with that in the spring of 2015, however, the diversity of food resources was higher in the summer of 2014.

Laminaria digitata as potential carbon source in heterotrophic microalgae cultivation for the production of fish feed supplement

A novel concept using the macroalgae Laminaria digitata as substrate to grow heterotrophically microalgae species to be used as fish feed supplement is investigated in the present study. Enzymatic hydrolysis of the macroalgae was performed to release the sugars present in the biomass. The hydrolysate was then used as culture medium and nutrient source to grow heterotrophically three different microalgae species, namely Chlorella protothecoides, Chlorella vulgaris and Chlorella sorokiniana, identified as valuable sources of proteins. Due to its faster growth rates, C. protothecoides was selected for further cultivation in batch reactors and its protein content and amino acid composition were measured. At the end of the process the biomass production reached 10.68±1.33gL−1 with a total protein accumulation of 41.77±1.82% (dry weight basis) and a protein yield of 0.17±0.06. Moreover, the essential amino acids score at the end of the experiment was 6 times greater than for the original content of the macroalgae hydrolysate which was used as substrate for the microalgae cultivation. Therefore, this study reveals the potential of macroalgae hydrolysate as culture medium for microalgae cultivation and it opens possibilities for the development of future strategies to optimize the microalgae production processes.

Electrochemical Upgrading of Bio-Oil

Bio-oil produced by fast pyrolysis of biomass is a potential source of low carbon, renewable hydrocarbon fuel. However, the properties such as low heating value, incomplete volatility, acidity, instability, and incompatibility with standard petroleum fuels significantly restrict its use as fuel. The undesirable properties of pyrolysis oil result from its chemical composition that mostly consists of different classes of oxygenated organic compounds. The elimination of oxygen is thus necessary to transform bio-oil into a liquid fuel that would be broadly accepted and economically attractive. Hydrodeoxygenation (HDO) or hydrotreating involves high-temperature, high-pressure processing in the presence of hydrogen and catalyst to remove oxygen. HDO suffers from significant challenges such as coking, deactivation of HDO catalysts by the presence of water in the pyrolysis oil, requirement of significant quantities of hydrogen to remove oxygen, and economic availability of hydrogen at a scale suitable for distributed biomass conversion. An alternative economically feasible, less hydrogen dependent and decentralized process is required to convert bio-oil to refinery ready hydrocarbons. A process of deoxygenation of bio-oil using solid-state, oxygen conductor based electrochemical cell is under investigation. The cell is operated at 500 – 550 °C to match the typical pyrolysis temperature for both physical and process integration of the two operations. The electrolysis process removes oxygen from the oxygenated organic molecule as well from steam to produce hydrogen in-situ. Thus, relatively small quantities of external hydrogen may be needed for deoxygenation, allowing for a distributed, small scale integrated upgrading unit. Mixtures of model compounds were tested using button cells and short stacks. The product from the electrochemical cell contained a suite of compounds with significantly lower oxygen content. Integrated testing of short stacks at the Pacific Northwest National Laboratory using a slip stream from a pyrolyzer shows that the product composition is similar to catalytic pyrolysis process. Additional tests are planned using improved electrode materials. Acknowledgment: This material is based upon work supported by the Department of Energy under Award Number DE-EE0006288.

Multicomponent quantification of Astragalus residue fermentation liquor using ion chromatography-integrated pulsed amperometric detection.

Chinese medicine residues contain abundant cellulose and hemicellulose, which are potential renewable carbon sources for ethanol production. The aim of the present study was to develop a rapid and reliable method to evaluate the cellulose and hemicellulose utilization in Chinese medicine residues. In the present study, key hydrolysates (arabinose, galactose, glucose, xylose, and cellobiose) of the cellulose and hemicellulose in fermentation liquor of Astragalus residues were simultaneously quantified by ion chromatography using an integrated pulsed amperometric detector (IPAD). HPLC analysis was performed on a Dionex ICS-2500 equipped with GP50 gradient pump and ED50 IPAD. The working and reference electrodes were gold electrode and Ag/AgCl electrode, respectively. Separation was achieved on serial no. 002627 Dionex Analytical column (2×250 mm). Sodium hydroxide of 250 mM and water were used as the mobile phase with a flow rate of 0.2 ml/min. The temperature of column was kept at 30°C. This method was validated for accuracy and precision. The regression equation revealed a good linear relationship (R2=0.9959–0.9984) within the test ranges. The limits of detection and quantification for five standard analytes (arabinose, galactose, glucose, xylose and cellobiose) were in the range of 0.067–0.091 and 0.08–0.23 mg/l, respectively. The method showed good reproducibility for the quantification of five analytes in fermentation liquor of Astragalus residue with intra-and inter-day variations less than 3.843%.

A molasses habitat-derived fungus Aspergillus tubingensis XG21 with high \u03b2-fructofuranosidase activity and its potential use for fructooligosaccharides production

The industrial microorganisms used for fructooligosaccharides (FOS) synthesis are generally fermented with sucrose as carbon source to induce the production of β-fructofuranosidase (FFase) having transfructosylation activity. Consequently, isolation of novel FFase producers from a sucrose-enriched biotope would help improve FOS productivity and reduce the process cost. Here, three fungi isolated from a unique sugarcane molasses habitat were found to possess FFase activity and one of them, XG21, exhibited a high capacity to synthesize FOS. Analysis of its morphological properties and ribosomal internal transcribed spacer (ITS) sequence allowed the taxonomic position to be assigned and it was thus identified as Aspergillus tubingensis XG21. It could utilize various potential carbon sources for vigorous growth, but only produced high-level FFase activity on sucrose. Furthermore, the transfructosylation ability and FOS synthesis were analyzed by TLC and HPLC. During the transfructosylation reaction, an increase in sucrose concentration led to the remarkable enhancement in FOS formation with the maximum content of up to 56.9% within 8 h. Finally, the sugarcane molasses was used to cultivate A. tubingensis XG21 and the optimal FFase activity reached up to 558.3 U/g, which was 88.9% higher than that with sucrose as carbon source. These results indicate that A. tubingensis XG21 can be considered as a new genetic resource adapted to cheaply available carbon sources for FOS production.