Browsing by Author "Zhang, Y. H. Percival"

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    Advances in Biochemical Engineering-Biotechnology
    Zhang, Y. H. Percival; Rollin, Joseph A.; Ye, Xinhao; Del Campo, Julia S. Martin; Adams, Michael W. W. (Springer, 2014-07-15)
    In vitro hydrogen generation represents a clear opportunity for novel bioreactor and system design. Hydrogen, already a globally important commodity chemical, has the potential to become the dominant transportation fuel of the future. Technologies such as in vitro synthetic pathway biotransformation (SyPaB)—the use of more than 10 purified enzymes to catalyze unnatural catabolic pathways—enable the storage of hydrogen in the form of carbohydrates. Biohydrogen production from local carbohydrate resources offers a solution to the most pressing challenges to vehicular and bioenergy uses: small-size distributed production, minimization of CO2 emissions, and potential low cost, driven by high yield and volumetric productivity. In this study, we introduce a novel bioreactor that provides the oxygen-free gas phase necessary for enzymatic hydrogen generation while regulating temperature and reactor volume. A variety of techniques are currently used for laboratory detection of biohydrogen, but the most information is provided by a continuous low-cost hydrogen sensor. Most such systems currently use electrolysis for calibration; here an alternative method, flow calibration, is introduced. This system is further demonstrated here with the conversion of glucose to hydrogen at a high rate, and the production of hydrogen from glucose 6-phosphate at a greatly increased reaction rate, 157 mmol/L/h at 60 [degrees] C.
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    Analysis of horse genomes provides insight into the diversification and adaptive evolution of karyotype
    Huang, J. L.; Zhao, Y. P.; Shiraigol, W.; Li, B.; Bai, D. Y.; Ye, W. X.; Daidiikhuu, D.; Yang, L. H.; Jin, Brqqg; Zhao, Q. A.; Gao, Y. H.; Wu, J.; Bao, Wydl; Li, A. A.; Zhang, Y. H. Percival; Han, H. G.; Bai, H. T.; Bao, Y. Q.; Zhao, L. L.; Zhai, Z. X.; Zhao, W. J.; Sun, Z. K.; Zhang, Y.; Meng, H.; Dugarjaviin, M. (Nature Publishing Group, 2014-05)
    Karyotypic diversification is more prominent in Equus species than in other mammals. Here, using next generation sequencing technology, we generated and de novo assembled quality genomes sequences for a male wild horse (Przewalski's horse) and a male domestic horse (Mongolian horse), with about 93-fold and 91-fold coverage, respectively. Portion of Y chromosome from wild horse assemblies (3 M bp) and Mongolian horse (2 M bp) were also sequenced and de novo assembled. We confirmed a Robertsonian translocation event through the wild horse's chromosomes 23 and 24, which contained sequences that were highly homologous with those on the domestic horse's chromosome 5. The four main types of rearrangement, insertion of unknown origin, inserted duplication, inversion, and relocation, are not evenly distributed on all the chromosomes, and some chromosomes, such as the X chromosome, contain more rearrangements than others, and the number of inversions is far less than the number of insertions and relocations in the horse genome. Furthermore, we discovered the percentages of LINE_L1 and LTR_ERV1 are significantly increased in rearrangement regions. The analysis results of the two representative Equus species genomes improved our knowledge of Equus chromosome rearrangement and karyotype evolution.
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    Cell-free protein synthesis energized by slowly-metabolized maltodextrin
    Wang, Yiran; Zhang, Y. H. Percival (2009-06-28)
    Background Cell-free protein synthesis (CFPS) is a rapid and high throughput technology for obtaining proteins from their genes. The primary energy source ATP is regenerated from the secondary energy source through substrate phosphorylation in CFPS. Results Distinct from common secondary energy sources (e.g., phosphoenolpyruvate - PEP, glucose-6-phosphate), maltodextrin was used for energizing CFPS through substrate phosphorylation and the glycolytic pathway because (i) maltodextrin can be slowly catabolized by maltodextrin phosphorylase for continuous ATP regeneration, (ii) maltodextrin phosphorylation can recycle one phosphate per reaction for glucose-1-phosphate generation, and (iii) the maltodextrin chain-shortening reaction can produce one ATP per glucose equivalent more than glucose can. Three model proteins, esterase 2 from Alicyclobacillus acidocaldarius, green fluorescent protein, and xylose reductase from Neurospora crassa were synthesized for demonstration. Conclusion Slowly-metabolized maltodextrin as a low-cost secondary energy compound for CFPS produced higher levels of proteins than PEP, glucose, and glucose-6-phospahte. The enhancement of protein synthesis was largely attributed to better-controlled phosphate levels (recycling of inorganic phosphate) and a more homeostatic reaction environment.
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    Cellulose-solvent-based lignocellulose fractionation with modest reaction conditions and reagent cycling
    (United States Patent and Trademark Office, 2014-07-22)
    Embodiments of the present invention overcome the well-known recalcitrance of lignocellulosic biomass in an economically viable manner. A process and system are provided for the efficient fractionation of lignocellulosic biomass into cellulose, hemicellulose sugars, lignin, and acetic acid. The cellulose thus obtained is highly amorphous and can be readily converted into glucose using known methods. Fermentable hemicellulose sugars, low-molecular-weight lignin, and purified acetic acid are also major products of the process and system. The modest process conditions and low solvent/solid ratios of some embodiments of the invention imply relatively low capital and processing costs.
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    Cellulose-solvent-based lignocellulose fractionation with modest reaction conditions and reagent cycling
    (United States Patent and Trademark Office, 2017-05-30)
    Embodiments of the present invention overcome the well-known recalcitrance of lignocellulosic biomass in an economically viable manner. A process and system are provided for the efficient fractionation of lignocellulosic biomass into cellulose, hemicellulose sugars, lignin, and acetic acid. The cellulose thus obtained is highly amorphous and can be readily converted into glucose using known methods. Fermentable hemicellulose sugars, low-molecular-weight lignin, and purified acetic acid are also major products of the process and system. The modest process conditions and low solvent/solid ratios of some embodiments of the invention imply relatively low capital and processing costs.
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    Coenzyme Engineering of a Hyperthermophilic 6-Phosphogluconate Dehydrogenase from NADP(+) to NAD(+) with Its Application to Biobatteries
    Chen, Hui; Zhu, Zhiguang; Huang, Rui; Zhang, Y. H. Percival (Nature Publishing Group, 2016-11-02)
    Engineering the coenzyme specificity of redox enzymes plays an important role in metabolic engineering, synthetic biology, and biocatalysis, but it has rarely been applied to bioelectrochemistry. Here we develop a rational design strategy to change the coenzyme specificity of 6-phosphogluconate dehydrogenase (6PGDH) from a hyperthermophilic bacterium Thermotoga maritima from its natural coenzyme NADP(+) to NAD(+). Through amino acid-sequence alignment of NADP(+)-and NAD(+)-preferred 6PGDH enzymes and computer-aided substrate-coenzyme docking, the key amino acid residues responsible for binding the phosphate group of NADP(+) were identified. Four mutants were obtained via site-directed mutagenesis. The best mutant N32E/R33I/T34I exhibited a x 6.4 x 10(4)-fold reversal of the coenzyme selectivity from NADP(+) to NAD(+). The maximum power density and current density of the biobattery catalyzed by the mutant were 0.135 mW cm(-2) and 0.255 mA cm(-2), similar to 25% higher than those obtained from the wide-type 6PGDH-based biobattery at the room temperature. By using this 6PGDH mutant, the optimal temperature of running the biobattery was as high as 65 degrees C, leading to a high power density of 1.75 mW cm(-2). This study demonstrates coenzyme engineering of a hyperthermophilic 6PGDH and its application to high-temperature biobatteries.
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    Development of Building Blocks - Thermostable Enzymes for Synthetic Pathway Biotransformation (SyPaB)
    Sun, Fangfang (Virginia Tech, 2012-04-25)
    Hydrogen production from abundant renewable biomass would decrease reliance on crude oils, achieve nearly zero net greenhouse gas emissions, create more jobs, and enhance national energy security. Cell-free synthetic pathway biotransformation (SyPaB) is the implementation of complicated chemical reaction by the in vitro assembly of numerous enzymes and coenzymes that microbes cannot do. One of the largest challenges is the high cost and instability of enzymes and cofactors. To overcome this obstacle, strong motivations have driven intensive efforts in discovering, engineering, and producing thermostable enzymes. In this project, ribose-5-phosphate isomerase (RpiB), one of the most important enzymes in the pentose phosphate pathway, was cloned from a thermophile Thermotoga maritima, and heterologously expressed in Escherichia coli, purified and characterized. High-purity RpiB was obtained by heat pretreatment through its optimization in buffer choice, buffer pH, as well as temperature and duration of pretreatment. This enzyme had the maximum activity at 80°C and pH 6.5-8.0. It had a half lifetime of 71 h at 60°C, resulting in its turn-over number of more than 2 x108 mol of product per mol of enzyme. Another two thermostable enzymes glucose-6-phosphate dehydrogenase (G6PDH) and diaphorase (DI) and their fusion proteins G6PDH-DI and DI-G6PDH were cloned from Geobacillus stearothermophilus, heterologouely expressed in E. coli and purified through its His-tag. The individual proteins G6PDH and DI have good thermostability and reactivity. However, the presence of DI in fusion proteins drastically decreased G6DPH activity. However, a mixture of G6PDH and a fusion protein G6PDH-DI not only restored G6PDH activity through the formation of heteromultimeric network but also facilitated substrate channeling between DI and G6PDH, especially at low enzyme concentrations. My researches would provide important building blocks for the on-going projects: high-yield hydrogen production through cell-free enzymatic pathways and electrical energy production through enzymatic fuel cells.
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    Doubling Power Output of Starch Biobattery Treated by the Most Thermostable Isoamylase from an Archaeon Sulfolobus tokodaii
    Cheng, Kun; Zhang, Fei; Sun, Fangfang; Chen, Hongge; Zhang, Y. H. Percival (Nature, 2015-08-20)
    Biobattery, a kind of enzymatic fuel cells, can convert organic compounds (e.g., glucose, starch) to electricity in a closed system without moving parts. Inspired by natural starch metabolism catalyzed by starch phosphorylase, isoamylase is essential to debranch alpha-1,6-glycosidic bonds of starch, yielding linear amylodextrin – the best fuel for sugar-powered biobattery. However, there is no thermostable isoamylase stable enough for simultaneous starch gelatinization and enzymatic hydrolysis, different from the case of thermostable alpha-amylase. A putative isoamylase gene was mined from megagenomic database. The open reading frame ST0928 from a hyperthermophilic archaeron Sulfolobus tokodaii was cloned and expressed in E. coli. The recombinant protein was easily purified by heat precipitation at 80 °C for 30 min. This enzyme was characterized and required Mg²⁺ as an activator. This enzyme was the most stable isoamylase reported with a half lifetime of 200 min at 90 °C in the presence of 0.5 mM MgCl₂, suitable for simultaneous starch gelatinization and isoamylase hydrolysis. The cuvett-based air-breathing biobattery powered by isoamylase-treated starch exhibited nearly doubled power outputs than that powered by the same concentration starch solution, suggesting more glucose 1-phosphate generated.
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    Energy Efficiency Analysis: Biomass-to-Wheel Efficiency Related with Biofuels Production, Fuel Distribution, and Powertrain Systems
    Huang, Wei-Dong; Zhang, Y. H. Percival (PLOS, 2011-07-13)
    Background Energy efficiency analysis for different biomass-utilization scenarios would help make more informed decisions for developing future biomass-based transportation systems. Diverse biofuels produced from biomass include cellulosic ethanol, butanol, fatty acid ethyl esters, methane, hydrogen, methanol, dimethyether, Fischer-Tropsch diesel, and bioelectricity; the respective powertrain systems include internal combustion engine (ICE) vehicles, hybrid electric vehicles based on gasoline or diesel ICEs, hydrogen fuel cell vehicles, sugar fuel cell vehicles (SFCV), and battery electric vehicles (BEV). Methodology/Principal Findings We conducted a simple, straightforward, and transparent biomass-to-wheel (BTW) analysis including three separate conversion elements -- biomass-to-fuel conversion, fuel transport and distribution, and respective powertrain systems. BTW efficiency is a ratio of the kinetic energy of an automobile's wheels to the chemical energy of delivered biomass just before entering biorefineries. Up to 13 scenarios were analyzed and compared to a base line case – corn ethanol/ICE. This analysis suggests that BEV, whose electricity is generated from stationary fuel cells, and SFCV, based on a hydrogen fuel cell vehicle with an on-board sugar-to-hydrogen bioreformer, would have the highest BTW efficiencies, nearly four times that of ethanol-ICE. Significance In the long term, a small fraction of the annual US biomass (e.g., 7.1%, or 700 million tons of biomass) would be sufficient to meet 100% of light-duty passenger vehicle fuel needs (i.e., 150 billion gallons of gasoline/ethanol per year), through up to four-fold enhanced BTW efficiencies by using SFCV or BEV. SFCV would have several advantages over BEV: much higher energy storage densities, faster refilling rates, better safety, and less environmental burdens.
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    A Hidden Transhydrogen Activity of a FMN-Bound Diaphorase under Anaerobic Conditions
    Collins, John; Zhang, Ting; Huston, Scott; Sun, Fangfang; Zhang, Y. H. Percival; Fu, Jinglin (PLOS, 2016-05-04)
    Background Redox cofactors of NADH/NADPH participate in many cellular metabolic pathways for facilitating the electron transfer from one molecule to another in redox reactions. Transhydrogenase plays an important role in linking catabolism and anabolism, regulating the ratio of NADH/NADPH in cells. The cytoplasmic transhydrogenases could be useful to engineer synthetic biochemical pathways for the production of high-value chemicals and biofuels. Methodology/Principal Findings A transhydrogenase activity was discovered for a FMN-bound diaphorase (DI) from Geobacillus stearothermophilus under anaerobic conditions. The DI-catalyzed hydride exchange were monitored and characterized between a NAD(P)H and a thio-modified NAD+ analogue. This new function of DI was demonstrated to transfer a hydride from NADPH to NAD+ that was consumed by NAD-specific lactate dehydrogenase and malic dehydrogenase. Conclusions/Significance We discover a novel transhydrogenase activity of a FMN-DI by stabilizing the reduced state of FMNH2 under anaerobic conditions. FMN-DI was demonstrated to catalyze the hydride transfer between NADPH and NAD+. In the future, it may be possible to incorporate this FMN-DI into synthetic enzymatic pathways for balancing NADH generation and NADPH consumption for anaerobic production of biofuels and biochemicals.
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    A high-energy-density sugar biobattery based on a synthetic enzymatic pathway
    Zhu, Zhiguang; Tam, Tsz Kin; Sun, Fangfang; You, Chun; Zhang, Y. H. Percival (Springer Nature, 2014-01)
    High-energy-density, green, safe batteries are highly desirable for meeting the rapidly growing needs of portable electronics. The incomplete oxidation of sugars mediated by one or a few enzymes in enzymatic fuel cells suffers from low energy densities and slow reaction rates. Here we show that nearly 24 electrons per glucose unit of maltodextrin can be produced through a synthetic catabolic pathway that comprises 13 enzymes in an air-breathing enzymatic fuel cell. This enzymatic fuel cell is based on non-immobilized enzymes that exhibit a maximum power output of 0.8 mW cm(-2) and a maximum current density of 6 mA cm(-2), which are far higher than the values for systems based on immobilized enzymes. Enzymatic fuel cells containing a 15% (wt/v) maltodextrin solution have an energy-storage density of 596 Ah kg(-1), which is one order of magnitude higher than that of lithium-ion batteries. Sugar-powered biobatteries could serve as next-generation green power sources, particularly for portable electronics.
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    High-Yield Cellulosic Hydrogen Production by Cell-Free Synthetic Cascade Enzymes: Minimal Bacterial Cellulase Cocktail and Thermostable Polyphosphate Glucokinase
    Liao, Hehuan (Virginia Tech, 2011-04-27)
    Hydrogen production from abundant renewable biomass would decrease reliance on crude oils, achieve nearly zero net greenhouse gas emissions, create more jobs, and enhance national energy security. Cell-free synthetic pathway biotransformation (SyPaB) is the implementation of complicated chemical reaction by the in vitro assembly of numerous enzymes and coenzymes. Two of the biggest challenges for its commercialization are: effective release of fermentable sugars from pretreated biomass, and preparations of thermostable enzymes with low-cost. The hydrolysis performance of 21 reconstituted bacterial cellulase mixtures containing the glycoside hydrolase family 5 Bacillus subtilis endoglucanase, family 9 Clostridium phytofermentans processive endoglucanase, and family 48 Clostridium phytofermentans cellobiohydrolase was investigated on microcrystalline cellulose (Avicel) and regenerated amorphous cellulose (RAC). The optimal ratios for maximum cellulose digestibility were dynamic for Avicel but nearly fixed for RAC. Processive endoglucanase CpCel9 was most important for high cellulose digestibility regardless of substrate type. These results suggested that the hydrolysis performance of reconstituted cellulase cocktail strongly depended on experimental conditions. Thermobifida fusca YX was hypothesized to have a thermophilic polyphosphate glucokinase. T. fusca YX ORF Tfu_1811 encoding a putative PPGK was cloned and the recombinant protein fused with a family 3 cellulose-binding module (CBM-PPGK) was over expressed in Escherichia coli. By a simple one-step immobilization, the half-life time increased to 2 h, at 50 °C. These results suggest that this enzyme was the most thermostable PPGK reported. My studies would provide important information for the on-going project: high-yield hydrogen production from cellulose by cell-free synthetic enzymatic pathway.
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    High-Yield Hydrogen Production from Starch and Water by a Synthetic Enzymatic Pathway
    Zhang, Y. H. Percival; Evans, Barbara R.; Mielenz, Jonathan R.; Hopkins, Robert C.; Adams, Michael W. W. (2007-05-23)
    Background. The future hydrogen economy offers a compelling energy vision, but there are four main obstacles: hydrogen production, storage, and distribution, as well as fuel cells. Hydrogen production from inexpensive abundant renewable biomass can produce cheaper hydrogen, decrease reliance on fossil fuels, and achieve zero net greenhouse gas emissions, but current chemical and biological means suffer from low hydrogen yields and/or severe reaction conditions. Methodology/Principal Findings. Here we demonstrate a synthetic enzymatic pathway consisting of 13 enzymes for producing hydrogen from starch and water. The stoichiometric reaction is C₆H₁₀O₅ (l)+7 H₂O (l)-> 12 H₂ (g)+ 6 CO₂ (g). The overall process is spontaneous and unidirectional because of a negative Gibbs free energy and separation of the gaseous products with the aqueous reactants. Conclusions. Enzymatic hydrogen production from starch and water mediated by 13 enzymes occurred at 30 degrees C as expected, and the hydrogen yields were much higher than the theoretical limit (4 H(2)/glucose) of anaerobic fermentations. Significance. The unique features, such as mild reaction conditions (30 degrees C and atmospheric pressure), high hydrogen yields, likely low production costs ($~ to 2/kg H₂), and a high energy-density carrier starch (14.8 H₂-based mass%), provide great potential for mobile applications. With technology improvements and integration with fuel cells, this technology also solves the challenges associated with hydrogen storage, distribution, and infrastructure in the hydrogen economy.
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    Investigating biomass saccharification for the production of cellulosic ethanol
    Zhu, Zhiguang (Virginia Tech, 2009-04-28)
    The production of second generation biofuels -- cellulosic ethanol from renewable lignocellulosic biomass has the potential to lead the bioindustrial revolution necessary to the transition from a fossil fuel-based economy to a sustainable carbohydrate economy. Effective release of fermentable sugars through biomass pretreatment followed by enzymatic hydrolysis is among the most costly steps for emerging cellulosic ethanol biorefineries. In this project, two pretreatment methods (dilute acid, DA, and cellulose solvent- and organic solvent-lignocellulose fractionation, COSLIF) for corn stover were compared. It was found that glucan digestibility of the corn stover pretreated by COSLIF was much higher, along with faster hydrolysis rate, than that by DA- pretreated. This difference was more significant at a low enzyme loading. Quantitative measurements of total substrate accessibility to cellulase (TSAC), cellulose accessibility to cellulase (CAC), and non-cellulose accessibility to cellulase (NCAC) based on adsorption of a non-hydrolytic recombinant protein TGC were established to find out the cause. The COSLIF-pretreated corn stover had a CAC nearly twice that of the DA-pretreated biomass. Further supported by qualitative scanning electron microscopy images, these results suggested that COSLIF treatment disrupted microfibrillar structures within biomass while DA treatment mainly removed hemicelluloses, resulting in a much less substrate accessibility of the latter than of the former. It also concluded that enhancing substrate accessibility was the key to an efficient bioconversion of lignocellulose. A simple method for determining the adsorbed cellulase on cellulosic materials or pretreated lignocellulose was established for better understanding of cellulase adsorption and desorption. This method involved hydrolysis of adsorbed cellulase in the presence of 10 M of NaOH at 121oC for 20 min, followed by the ninhydrin assay for the amino acids released from the hydrolyzed cellulase. The major lignocellulosic components (i.e. cellulose, hemicellulose, and lignin) did not interfere with the ninhydrin assay. A number of cellulase desorption methods were investigated, including pH adjustment, detergents, high salt solution, and polyhydric alcohols. The pH adjustment to 13.0 and the elution by 72% ethylene glycol at a neutral pH were among the most efficient approaches for desorbing the adsorbed cellulase. For the recycling of active cellulase, a modest pH adjustment to 10.0 may be a low-cost method to desorb active cellulase. More than 90% of cellulase for hydrolysis of the pretreated corn stover could be recycled by washing at pH 10.0. This study provided an in-depth understanding of biomass saccharification for the production of cellulosic ethanol for cellulose hydrolysis and cellulase adsorption and desorption. It will be of great importance for developing better lignocellulose pretreatment technologies and improving cellulose hydrolysis by engineered cellulases.
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    Method and apparatus for lignocellulose pretreatment using a super-cellulose-solvent and highly volatile solvents
    (United States Patent and Trademark Office, 2014-12-02)
    Embodiments of the present invention overcome the well-known recalcitrance of lignocellulosic biomass in an economically viable manner. A process and a system are provided for the efficient fractionation of lignocellulosic biomass into cellulose, hemicellulose, and lignin. The cellulose and hemicellulose thus obtained are highly amorphous and can be readily converted into highly concentrated mixtures of five and six carbon sugars using known methods. Typical yields of sugars exceed 100 grams of sugars per liter of sugar solution. Other products, such as alcohols, can easily be prepared according to methods of the invention. The modest process conditions and low solvent/solid ratios of some embodiments of the invention require relatively low capital and processing costs.
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    Method and apparatus for lignocellulose pretreatment using a super-cellulose-solvent and highly volatile solvents
    (United States Patent and Trademark Office, 2014-03-04)
    Embodiments of the present invention overcome the well-known recalcitrance of lignocellulosic biomass in an economically viable manner. A process and a system are provided for the efficient fractionation of lignocellulosic biomass into cellulose, hemicellulose, and lignin. The cellulose and hemicellulose thus obtained are highly amorphous and can be readily converted into highly concentrated mixtures of five and six carbon sugars using known methods. Typical yields of sugars exceed 100 grams of sugars per liter of sugar solution. Other products, such as alcohols, can easily be prepared according to methods of the invention. The modest process conditions and low solvent/solid ratios of some embodiments of the invention require relatively low capital and processing costs.
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    New artificial fluoro-cofactor of hydride transfer with novel fluorescence assay for redox biocatalysis
    Zhang, Lei; Yuan, Jun; Xu, Y.; Zhang, Y. H. Percival; Qian, Xuhong (Royal Society of Chemistry, 2016-01-01)
    A new artificial fluoro-cofactor was developed for the replacement of natural cofactors NAD(P), exhibiting a high hydride transfer ability. More importantly, we established a new and fast screening method for the evaluation of the properties of artificial cofactors based on the fluorescence assay and visible color change.
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    Non-Complexed Four Cascade Enzyme Mixture: Simple Purification and Synergetic Co-stabilization
    Myung, Suwan; Zhang, Y. H. Percival (PLoS, 2013-04-09)
    Cell-free biosystems comprised of synthetic enzymatic pathways would be a promising biomanufacturing platform due to several advantages, such as high product yield, fast reaction rate, easy control and access, and so on. However, it was essential to produce (purified) enzymes at low costs and stabilize them for a long time so to decrease biocatalyst costs. We studied the stability of the four recombinant enzyme mixtures, all of which originated from thermophilic microorganisms: triosephosphate isomerase (TIM) from Thermus thermophiles, fructose bisphosphate aldolase (ALD) from Thermotoga maritima, fructose bisphosphatase (FBP) from T. maritima, and phosphoglucose isomerase (PGI) from Clostridium thermocellum. It was found that TIM and ALD were very stable at evaluated temperature so that they were purified by heat precipitation followed by gradient ammonia sulfate precipitation. In contrast, PGI was not stable enough for heat treatment. In addition, the stability of a low concentration PGI was enhanced by more than 25 times in the presence of 20 mg/L bovine serum albumin or the other three enzymes. At a practical enzyme loading of 1000 U/L for each enzyme, the half-life time of free PGI was prolong to 433 h in the presence of the other three enzymes, resulting in a great increase in the total turn-over number of PGI to 6.2×109 mole of product per mole of enzyme. This study clearly suggested that the presence of other proteins had a strong synergetic effect on the stabilization of the thermolabile enzyme PGI due to in vitro macromolecular crowding effect. Also, this result could be used to explain why not all enzymes isolated from thermophilic microorganisms are stable in vitro because of a lack of the macromolecular crowding environment.
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    Observation of Electron-Antineutrino Disappearance at Daya Bay
    An, F. P.; Bai, J. Z.; Balantekin, A. B.; Band, H. R.; Beavis, D.; Beriguete, W.; Bishai, M.; Blyth, S.; Boddy, K.; Brown, R. L.; Cai, B.; Cao, G. F.; Cao, J.; Carr, Rachel E.; Chan, W. T.; Chang, J. F.; Chang, Y.; Chasman, C.; Chen, H. S.; Chen, H. Y.; Chen, S. J.; Chen, S. M.; Chen, X. C.; Chen, X. H.; Chen, X. S.; Chen, Y.; Chen, Y. X.; Cherwinka, J. J.; Chu, M. C.; Cummings, J. P.; Deng, Z. Y.; Ding, Y. Y.; Diwan, M. V.; Dong, L.; Draeger, E.; Du, X. F.; Dwyer, D. A.; Edwards, W. R.; Ely, S. R.; Fang, S. D.; Fu, J. Y.; Fu, Z. W.; Ge, L. Q.; Ghazikhanian, V.; Gill, R. L.; Goett, J.; Gonchar, M.; Gong, G. H.; Gong, H.; Gornushkin, Y. A.; Greenler, L. S.; Gu, W. Q.; Guan, M. Y.; Guo, X. H.; Hackenburg, R. W.; Hahn, R. L.; Hans, S.; He, M.; He, Q.; He, W. S.; Heeger, K. M.; Heng, Y. K.; Hinrichs, P.; Ho, T. H.; Hor, Y. K.; Hsiung, Y. B.; Hu, B. Z.; Hu, T.; Huang, H. X.; Huang, H. Z.; Huang, P. W.; Huang, X.; Huang, X. T.; Huber, Patrick; Isvan, Z.; Jaffe, D. E.; Jetter, S.; Ji, X. L.; Ji, X. P.; Jiang, H. J.; Jiang, W. Q.; Jiao, J. B.; Johnson, R. A.; Kang, L.; Kettell, S. H.; Kramer, M.; Kwan, K. K.; Kwok, M. W.; Kwok, T.; Lai, C. Y.; Lai, W. C.; Lai, W. H.; Lau, K.; Lebanowski, L.; Lee, J.; Lee, M. K. P.; Leitner, R.; Leung, J. K. C.; Leung, K. Y.; Lewis, C. A.; Li, B.; Li, F.; Li, G. S.; Li, J.; Li, Q. J.; Li, S. F.; Li, W. D.; Li, X. B.; Li, X. N.; Li, X. Q.; Li, Y.; Li, Z. B.; Liang, H.; Liang, J.; Lin, C. J.; Lin, G. L.; Lin, S. K.; Lin, S. X.; Lin, Y. C.; Ling, J. J.; Link, Jonathan M.; Littenberg, L.; Littlejohn, B. R.; Liu, B. J.; Liu, C.; Liu, D. W.; Liu, H.; Liu, J. C.; Liu, J. L.; Liu, S.; Liu, X.; Liu, Y. B.; Lu, C.; Lu, H. Q.; Luk, A.; Luk, K. B.; Luo, T.; Luo, X. L.; Ma, L. H.; Ma, Q. M.; Ma, X. B.; Ma, X. Y.; Ma, Y. Q.; Mayes, B.; McDonald, K. T.; McFarlane, M. C.; McKeown, R. D.; Meng, Y.; Mohapatra, D.; Morgan, J. E.; Nakajima, Y.; Napolitano, J.; Naumov, D.; Nemchenok, I.; Newsom, C.; Ngai, H. Y.; Ngai, W. K.; Nie, Y. B.; Ning, Z.; Ochoa-Ricoux, J. P.; Oh, D.; Olshevski, A.; Pagac, A.; Patton, S.; Pearson, C.; Pec, V.; Peng, J. C.; Piilonen, Leo E.; Pinsky, L.; Pun, C. S. J.; Qi, F. Z.; Qi, M.; Qian, X.; Raper, N.; Rosero, R.; Roskovec, B.; Ruan, X. C.; Seilhan, B.; Shao, B. B.; Shih, K.; Steiner, H.; Stoler, P.; Sun, G. X.; Sun, J. L.; Tam, Y. H.; Tanaka, H. K.; Tang, X.; Themann, H.; Torun, Y.; Trentalange, S.; Tsai, O.; Tsang, K. V.; Tsang, R. H. M.; Tull, C.; Viren, B.; Virostek, S.; Vorobel, V.; Wang, C. H.; Wang, L. S.; Wang, L. Y.; Wang, L. Z.; Wang, M.; Wang, N. Y.; Wang, R. G.; Wang, T.; Wang, W.; Wang, X.; Wang, Y. F.; Wang, Z.; Wang, Z. M.; Webber, D. M.; Wei, Y. D.; Wen, L. J.; Wenman, D. L.; Whisnant, K.; White, C. G.; Whitehead, L.; Whitten, C. A.; Wilhelmi, J.; Wise, T.; Wong, H. C.; Wong, H. L. H.; Wong, J.; Worcester, E.; Wu, F. F.; Wu, Q.; Xia, D. M.; Xiang, S. T.; Xiao, Q.; Xing, Z. Z.; Xu, G.; Xu, J.; Xu, J. L.; Xu, W.; Xu, Y.; Xue, T.; Yang, C. G.; Yang, L.; Ye, M.; Yeh, M.; Yeh, Y. S.; Yip, K.; Young, B. L.; Yu, Z. Y.; Zhan, L.; Zhang, C.; Zhang, F. H.; Zhang, J. W.; Zhang, Q. M.; Zhang, K.; Zhang, Q. X.; Zhang, S. H.; Zhang, Y. C.; Zhang, Y. H. Percival; Zhang, Y. X.; Zhang, Z. J.; Zhang, Z. P.; Zhang, Z. Y.; Zhao, J.; Zhao, Q. W.; Zhao, Y. B.; Zheng, L.; Zhong, W. L.; Zhou, L.; Zhou, Z. Y.; Zhuang, H. L.; Zou, J. H. (American Physical Society, 2012-04-23)
    The Daya Bay Reactor Neutrino Experiment has measured a nonzero value for the neutrino mixing angle 0(13) with a significance of 5.2 standard deviations. Antineutrinos from six 2.9 GW(th) reactors were detected in six antineutrino detectors deployed in two near (flux-weighted baseline 470 m and 576 m) and one far (1648 m) underground experimental halls. With a 43 000 ton-GW(th)-day live-time exposure in 55 days, 10 416 (80 376) electron-antineutrino candidates were detected at the far hall (near halls). The ratio of the observed to expected number of antineutrinos at the far hall is R = 0.940 +/- 0.011(stat.) +/- 0.004(syst.). A rate-only analysis finds sin(2)2 theta(13) = 0.092 +/- 0.016(stat.) +/- 0.005(syst.) in a three-neutrino framework.
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    Overcoming Biomass Recalcitrance by Combining Genetically Modified Switchgrass and Cellulose Solvent-Based Lignocellulose Pretreatment
    Sathitsuksanoh, Noppadon; Xu, Bin; Zhao, Bingyu Y.; Zhang, Y. H. Percival (2013-09-27)
    Decreasing lignin content of plant biomass by genetic engineering is believed to mitigate biomass recalcitrance and improve saccharification efficiency of plant biomass. In this study, we compared two different pretreatment methods (i.e., dilute acid and cellulose solvent) on transgenic plant biomass samples having different lignin contents and investigated biomass saccharification efficiency. Without pretreatment, no correlation was observed between lignin contents of plant biomass and saccharification efficiency. After dilute acid pretreatment, a strong negative correlation between lignin content of plant samples and overall glucose release was observed, wherein the highest overall enzymatic glucan digestibility was 70% for the low-lignin sample. After cellulose solvent- and organic solvent-based lignocellulose fractionation pretreatment, there was no strong correlation between lignin contents and high saccharification efficiencies obtained (i.e., 80-90%). These results suggest that the importance of decreasing lignin content in plant biomass to saccharification was largely dependent on pretreatment choice and conditions.