Optimum alcohol concentration for chain elongation in mixed-culture fermentation of cellulosic substrate.

Medium-chain fatty acids (MCFA, e.g., caproic, heptanoic, caprylic acid) are more valuable than short-chain fatty acids (SCFA, e.g., acetic, propionic, butyric, valeric acid). SCFAs are major products in methane-inhibited mixed-culture anaerobic fermentation. By feeding ethanol to the fermentor, MCFA formation is enhanced through chain elongation. Microorganisms such as Clostridium kluyveri elongate short-chain acids by combining them with alcohol. Very low ethanol concentration reduces chain elongation rates, whereas very high ethanol concentrations inhibit microorganisms. To maximize MCFA production, different ethanol concentrations were investigated in the mixed-culture fermentation of office paper and chicken manure. At 10 g/L ethanol concentration, 10 g/L MCFA was formed. High ethanol concentrations (above 40 g/L) inhibit microorganisms resulting in no chain elongation. For chain elongation, propanol was found to be more inhibitory than ethanol. The data suggest that MCFA production will increase by continuously extracting MCFA and maintaining 5-10 g/L ethanol concentration by periodic addition. Biotechnol. Bioeng. 2016;113: 2597-2604. © 2016 Wiley Periodicals, Inc.

Short-term oxidative lime pretreatment of palm pruning waste for use as animal feedstuff.

BACKGROUND: Oxidative lime pretreatment (OLP) is an effective pretreatment for highly recalcitrant lignocellulosic materials. This experiment was conducted to investigate the effect of short-term OLP on fermentative gas production kinetics of date palm prunings. Rachis and petiole were pretreated with excess lime (0.5 g Ca(OH)2 g(-1) dry matter) in a reactor charged with 10 bar pure oxygen pressure at different times and temperatures. RESULTS: Lignin removal was greatly affected by OLP, whereas cellulose was well preserved even after severe pretreatment. After 72 h fermentation, the cumulative gas production was 321.2 mL gas g(-1) organic matter (OM) for the most severe pretreatment, compared to 73.6 mL g(-1) OM for the untreated rachis. For the petiole pretreated at 120 °C for 280 min, 268 mL gas was produced compared to 59 mL gas g(-1) OM for the untreated petiole. Scanning electron microscope images showed the formation of pores (average diameter of 10-12 µm) and carbonate calcium deposits on the surface of treated biomass. An increase in biomass crystallinity was observed in pretreated samples resulting from cellulose enrichment. CONCLUSIONS: The results suggest that OLP improves the ruminal digestibility of date palm prunings, which may have potential for inclusion in the ruminant diet at low cost.

The Impact of Preservation Techniques on Methane-Arrested Anaerobic Digestion of Nutrient-Rich Feedstocks.

The carboxylate platform is a promising biomass-to-energy pathway that uses methane-arrested anaerobic digestion (MAAD) to convert biomass to carboxylic acids, which can be chemically converted to industrial chemicals and liquid fuels. Lignocellulose is an energy-rich carbon source, but lacks nutrients necessary for microbial growth. Chicken manure (rural waste) and sewage sludge (urban waste) are rich in nitrogen and useful macronutrients; therefore, co-digesting these wastes with lignocellulose improves MAAD performance. However, waste nutrients must be digested immediately, or preserved. This study investigated the effects of various preservation techniques - frozen (fresh), air-dried, and baked - on chicken manure and sewage sludge. Batch experiments were performed with office paper (carbon source) and chicken manure or sewage sludge (nutrient source) with different methods of preservation. Fresh substrates produced higher acid yields and biomass conversion (the amount of biomass consumed during digestion) than dried substrates. Baked chicken manure showed reduced conversion and total acid production, which suggests that oven-drying reduces digestibility. From the batch data, the Continuum Particle Distribution Model (CPDM) predicted results of a four-stage countercurrent digestion. The data are displayed on maps showing the impact of liquid residence time (LRT) and volatile solids loading rate (VSLR) on conversion and product concentration. Co-digesting office paper and wet chicken manure at a non-acid volatile solid (NAVS) concentration of 300 g/Lliq, the model predicted a high total acid concentration of 52.8 g/L and conversion of 0.89 g NAVSdigested/NAVSfed at a volatile solid loading rate of 4 g/(Lliq·day) and liquid retention time of 35 days.

Valorizing prickly pear cladodes via methane-arrested anaerobic digestion for carboxylic acid production.

To address climate change, liquid biofuels are an essential alternative to fossil fuels, especially for transportation. The carboxylate platform uses methane-arrested anaerobic digestion (MAAD) to ferment biomass to carboxylic acids, which can be chemically converted to liquid fuels via the carboxylate platform. Most biomass sources require expensive pretreatments to remove lignin; however, prickly pear (Opuntia ficus-indica) cladodes have low lignin content and therefore do not require pretreatment. Furthermore, this sugar-rich feedstock is readily digested to high concentrations of carboxylic acids. At various substrate concentrations, batch MAAD of prickly pear cladodes yielded primarily acetic, butyric, and caproic acids. From these batch data, continuum particle distribution modeling (CPDM) simulated four-stage countercurrent digestion. At a non-acid volatile solid (NAVS) concentration of 100 g/Lliq , CPDM predicts a high total acid concentration of 93 g/L and conversion of 0.93 g NAVSdigested /NAVSfed at a volatile solid loading rate of 6 g/(Lliq ·d) and liquid retention time of 35 days. Without chemical pretreatment, co-digestion, or in situ product removal, prickly pear produced high yields, biomass conversion, product concentration, and selectivity compared to previously studied lignocellulosic feedstocks.

Conversion of acetone and mixed ketones to hydrocarbons using HZSM-5 catalyst in the carboxylate platform.

In this study, two different feeds were treated to produce hydrocarbons: (1) reagent-grade acetone, and (2) mixed ketones obtained from lignocellulosic biomass via the carboxylate platform. Acetone and mixed ketones underwent catalytic self-condensation over HZSM-5. For acetone, HZSM-5(80) was used, and the experiments were conducted in two sets: (1) vary temperature (305-415°C) at P = 101 kPa (abs) and weight hourly space velocity (WHSV) = 1.3 h-1; (2) vary WHSV (1.3-7.9 h-1) at T = 350 and 415°C, and P = 101 kPa (abs). For acetone over HZSM-5(280), the experiments were conducted in two sets: (1) vary WHSV (1.3-6.5 h-1) at T = 415°C, and P = 101 kPa (abs); and (2) vary WHSV (1.3-11.8 h-1) at P = 790 kPa (abs) and T = 415°C. For mixed ketones, HZSM-5(280) was used at WHSV = 1.9 h-1, T = 430-590°C, and P = 101 kPa (abs). For acetone at higher temperatures, the conversion was 100% and the liquid products were aromatics centered on C8. At low temperatures, conversion was less and the carbon liquid distribution was centered on C9 (mainly mesitylene). For mixed ketones, catalyst deactivation was higher causing product concentrations to change over time, and the highest conversion reached was 40%.

Assessment of shock pretreatment and alkali pretreatment on corn stover using enzymatic hydrolysis.

This study investigates digestibility enhancements of lignocellulose from shock pretreatment, alkaline pretreatment, and combination. Shock pretreatment subjects aqueous slurries of lignocellulose to shock waves, which disrupts its structure rendering it more susceptible to hydrolysis. Alkaline pretreatment submerges the biomass in aqueous alkali (NaOH, Ca(OH)2 ), which removes lignin and acetyl groups. As indicators of digestibility, cellulase (CTec3) and hemicellulase (HTec3) were used to saccharify the pretreated corn stover and the resulting filtrate which contains about 10% of the sugars. Shock is most effective when it precedes alkaline pretreatment, presumably because it opens the biomass structure and enhances diffusion of pretreatment chemicals. Lignocellulose digestibility from calcium hydroxide treatment improves significantly with oxygen addition. In contrast, sodium hydroxide is a more potent alkali, and thereby eliminates the need for oxygen to enhance pretreatment. At low hydroxide loadings (<4 g OH- /100 g dry biomass), both NaOH and Ca(OH)2 provide similar increases in digestibility; however, at high hydroxide loadings, NaOH is superior. For animal feed, Ca(OH)2 treatment is recommended, because residual calcium ions are valuable nutrients. In contrast, for methane-arrested anaerobic digestion, NaOH treatment is preferred because NaHCO3 is a stronger buffer. At 50°C, shock pretreatment improves sugar yields at all NaOH loadings. The effect of shock is most pronounced when the no-shock control employed the same soaking-and-drying procedure as the shock treatment. The recommended conditions are shock treatment (5.52 bar [abs] initial H2 /O2 pressure) followed by 50°C alkaline treatment with NaOH loading of 4 g OH- /100 g dry biomass for 1 h.

Assessment of corn stover pretreated with shock and alkali using methane-arrested anaerobic digestion.

Corn stover, an underutilized agricultural residue, is a promising lignocellulosic feedstock for producing biofuels. To fully utilize it, pretreatment is needed. Typically, pretreatments are rapidly assessed using extracellular enzymes that release sugars from cellulose and hemicellulose. In contrast, this study uses methane-arrested anaerobic digestion (MAAD) to assess pretreatments. Although time consuming, MAAD is a more accurate assessment technique when lignocellulose is employed in the carboxylate platform, a promising approach that utilizes nearly all biomass components. Using recommended pretreatment conditions identified from a previous study, three corn stover pretreatments were compared using MAAD: (1) shock-only, (2) NaOH-only, and (3) shock + NaOH. Air-dried sewage sludge was used as nutrient source. At 100 g/L initial substrate concentration, compared to untreated corn stover, shock-only decreased conversion (amount of biomass digested) by 14%, NaOH-only increased conversion by 82%, and shock + NaOH increased conversion by 104%. Using batch MAAD data, the continuum particle distribution model simulated four-stage countercurrent fermentation. At an industrial non-acid volatile solids (NAVS) concentration of 300 g/Lliq , for both NaOH-only and shock + NaOH, the model predicts total carboxylic acid concentration of about 58 g/L and conversion of about 0.85 g NAVSdigested /g NAVSfed at liquid retention time of 35 days and volatile solid loading rate of 4 g/(Lliq ⋅day). At this long solid residence time, shock is not necessary; however, with short solid residence times, shock acts synergistically to aid NaOH pretreatment. Shock treatment offers a way to reduce pretreatment costs without sacrificing pretreatment efficacy.

Microbial communities for valorizing biomass using the carboxylate platform to produce volatile fatty acids: A review.

The carboxylate platform employs a diverse microbial consortium of anaerobes in which the methanogens are inhibited. Nearly all biomass components are digested to a mixture of C1-C8 monocarboxylic acids and their corresponding salts. The methane-arrested anaerobic digestion proceeds readily without needing to sterilize biomass or equipment. It accepts a wide range of feedstocks (e.g., agricultural residues, municipal solid waste, sewage sludge, animal manure, food waste, algae, and energy crops), and produces high product yields. This review highlights several important aspects of the platform, including its thermodynamic underpinnings, influences of inoculum source and operating conditions on product formation, and downstream chemical processes that convert the carboxylates to hydrocarbon fuels and oxygenated chemicals. This review further establishes the carboxylate platform as a viable and economical route to industrial biomass utilization.

Anaerobic thermophilic fermentation for carboxylic acid production from in-storage air-lime-treated sugarcane bagasse.

Wet storage and in situ lime pretreatment (50 °C, 1-atm air, 56 days, excess lime loading of 0.3 g Ca(OH)(2)/g dry biomass) of sugarcane bagasse (4,000 g dry weight) was performed in a bench-scale pile pretreatment system. Under thermophilic conditions (55 °C, NH(4)HCO(3) buffer, methane inhibitors), air-lime-treated bagasse (80 wt.%) and chicken manure (20 wt.%) were anaerobically co-digested in 1-L rotary fermentors by a mixed culture of marine microorganisms (Galveston, TX). During four-stage countercurrent fermentation, the resulting carboxylic acids consisted of primarily acetate (average 87.7 wt.%) and butyrate (average 9.0 wt.%). The experimental fermentation trains had the highest yield (0.47 g total acids/g volatile solids (VS) fed) and highest selectivity (0.79 g total acids/g VS digested) at a total acid concentration of 28.3 g/L, which is equivalent to an ethanol yield of 105.2 gal/(tonne VS fed). Both high total acid concentrations (>44.7 g/L) and high substrate conversions (>77.5%) are predicted for countercurrent fermentations of bagasse at commercial scale, allowing for an efficient conversion of air-lime-treated biomass to liquid transportation fuels and chemicals via the carboxylate platform.

Arrested methanogenesis digestion of high-strength cheese whey and brewery wastewater with carboxylic acid production.

A new anaerobic digestion process based on arrested methanogenesis (AM) was developed to treat high-strength cheese whey and brewery wastewater with simultaneous carboxylic acid production. This study specifically determined the links between wastewater characteristics, microbial community structure, and the operation of AM digesters at the bench scale. The highest total carboxylic acids concentration (78 g/L) was achieved after 15 days under batch condition at 40 °C and near-neutral pH. Lactate conversion to chain-elongated volatile fatty acid was observed. Under fed-batch conditions, the highest total acid productivity was 16 g/(Lliq·d) with substrate conversion of 0.66 g CODdigested/g CODfed at hydraulic residence time (HRT) of 4 days. Fed-batch digestion with biomass recycling resulted in a 2-fold increase in VFAs concentration (30 g/L) and a higher diversity in the microbial consortia. Experimental results show that highly efficient, robust, and productive community structure was established for sustainable carboxylate production from widely varying high-strength wastewaters.

Anaerobic mixed-culture fermentation of aqueous ammonia-treated sugarcane bagasse in consolidated bioprocessing.

The MixAlco process is an example of consolidated bioprocessing (CBP) in which anaerobic mixed-culture fermentation biochemically converts any biodegradable feedstock into carboxylate salts. Downstream processing thermochemically transforms the resulting salts into mixed alcohol fuels or gasoline. To enhance digestibility, sugarcane bagasse was treated under mild conditions (55 degrees C, 24 h, and 30% aqueous ammonia solution with a loading of 10 mL/g dry biomass). Using NH(4)HCO(3) buffer, the feedstock (80% ammonia-treated sugarcane bagasse/20% chicken manure) was anaerobically fermented by a mixed culture of marine microorganisms at 55 degrees C. Four-stage countercurrent fermentations were performed at various volatile solids loading rates (VSLRs) and liquid residence times (LRTs). The highest acid productivity (1.14 g/(L day)) occurred at a total acid concentration of 29.8 g/L. The highest conversion (65%) occurred at a total acid concentration of 27.6 g/L. The continuum particle distribution model (CPDM) predicted the experimental total acid concentrations and conversions within 4.98% and 10.41%, respectively. When using NH(4)HCO(3) buffer, ammonia pretreatment is an attractive option. The CPDM "map" shows that both high volatile solid conversions (78.8%) and high acid concentrations (32.6 g/L) are possible with 300 g/(L liquid) substrate concentration, 30 days LRT, 2 g/(L day) solid loading rate and NH(4)HCO(3) buffer.

Structure and dynamics of the microbial communities underlying the carboxylate platform for biofuel production.

The carboxylate platform utilizes a mixed microbial community to convert lignocellulosic biomass into chemicals and fuels. While much of the platform is well understood, little is known about its microbiology. Mesophilic (40 degrees C) and thermophilic (55 degrees C) fermentations employing a sorghum feedstock and marine sediment inoculum were profiled using 16S rRNA tag-pyrosequencing over the course of a 30-day incubation. The contrasting fermentation temperatures converted similar amounts of biomass, but the mesophilic community was significantly more productive, and the two temperatures differed significantly with respect to propionic and butyric acid production. Pyrotag sequencing revealed the presence of dynamic communities that responded rapidly to temperature and changed substantially over time. Both temperatures were dominated by bacteria resembling Clostridia, but they shared few taxa in common. The species-rich mesophilic community harbored a variety of Bacteroidetes, Actinobacteria, and gamma-Proteobacteria, whereas the thermophilic community was composed mainly of Clostridia and Bacilli. Despite differences in composition and productivity, similar patterns of functional class dynamics were observed. Over time, organisms resembling known cellulose degraders decreased in abundance, while organisms resembling known xylose degraders increased. Improved understanding of the carboxylate platform's microbiology will help refine platform performance and contribute to our growing knowledge regarding biomass conversion and biofuel production processes.

Techno-economic analysis of biomass to fuel conversion via the MixAlco process.

MixAlco is a robust process that converts biomass to fuels and chemicals. A key feature of the MixAlco process is the fermentation, which employs a mixed culture of acid-forming microorganisms to convert biomass components (carbohydrates, proteins, and fats) to carboxylate salts. Subsequently, these intermediate salts are chemically converted to hydrocarbon fuels (gasoline, jet fuel, and diesel). This work focuses on process synthesis, simulation, integration, and cost estimation of the MixAlco process. For the base-case capacity of 40 dry tonne feedstock per hour, the total capital investment is US $5.54/annual gallon of hydrocarbon fuels (US $3.79/annual gallon of ethanol equivalent), and the minimum selling price [with 10% return on investment (ROI), internal hydrogen production, and US $60/tonne biomass] is US $2.56/gal hydrocarbon, which is equivalent to US $1.75/gal ethanol. If plant capacity is increased to 400 tph, the minimum selling price of biomass-derived hydrocarbon fuels is US $1.76/gal hydrocarbon (US $1.20/gal ethanol equivalent), which can compete without subsidies with petroleum-derived hydrocarbons when crude oil sells for about US $65/bbl. At 40 tph, using the average tipping fee for municipal solid waste (US $45/dry tonne) and current price of external hydrogen (US $1/kg), the minimum selling price is only US $1.24/gal hydrocarbon (US $0.85/gal ethanol equivalent).

Kinetic modeling of countercurrent saccharification.

BACKGROUND: Countercurrent saccharification is a promising way to minimize enzyme loading while obtaining high conversions and product concentrations. However, in countercurrent saccharification experiments, 3-4 months are usually required to acquire a single steady-state data point. To save labor and time, simulation of this process is necessary to test various reaction conditions and determine the optimal operating point. Previously, a suitable kinetic model for countercurrent saccharification has never been reported. The Continuum Particle Distribution Modeling (CPDM) satisfactorily predicts countercurrent fermentation using mixed microbial cultures that digest various feedstocks. Here, CPDM is applied to countercurrent enzymatic saccharification of lignocellulose. RESULTS: CPDM was used to simulate multi-stage countercurrent saccharifications of a lignocellulose model compound (α-cellulose). The modified HCH-1 model, which accurately predicts long-term batch saccharification, was used as the governing equation in the CPDM model. When validated against experimental countercurrent saccharification data, it predicts experimental glucose concentrations and conversions with the average errors of 3.5% and 4.7%, respectively. CPDM predicts conversion and product concentration with varying enzyme-addition location, total stage number, enzyme loading, liquid residence time (LRT), and solids loading rate (SLR). In addition, countercurrent saccharification was compared to batch saccharification at the same conversion, product concentration, and reactor volume. Results show that countercurrent saccharification is particularly beneficial when the product concentration is low. CONCLUSIONS: The CPDM model was used to simulate multi-stage countercurrent saccharification of α-cellulose. The model predictions agreed well with the experimental glucose concentrations and conversions. CPDM prediction results showed that the enzyme-addition location, enzyme loading, LRT, and SLR significantly affected the glucose concentration and conversion. Compared to batch saccharification at the same conversion, product concentration, and reactor volume, countercurrent saccharification is particularly beneficial when the product concentration is low.

Development of modified HCH-1 kinetic model for long-term enzymatic cellulose hydrolysis and comparison with literature models.

BACKGROUND: Enzymatic hydrolysis is a major step for cellulosic ethanol production. A thorough understanding of enzymatic hydrolysis is necessary to help design optimal conditions and economical systems. The original HCH-1 (Holtzapple-Caram-Humphrey-1) model is a generalized mechanistic model for enzymatic cellulose hydrolysis, but was previously applied only to the initial rates. In this study, the original HCH-1 model was modified to describe integrated enzymatic cellulose hydrolysis. The relationships between parameters in the HCH-1 model and substrate conversion were investigated. Literature models for long-term (> 48 h) enzymatic hydrolysis were summarized and compared to the modified HCH-1 model. RESULTS: A modified HCH-1 model was developed for long-term (> 48 h) enzymatic cellulose hydrolysis. This modified HCH-1 model includes the following additional considerations: (1) relationships between coefficients and substrate conversion, and (2) enzyme stability. Parameter estimation was performed with 10-day experimental data using α-cellulose as substrate. The developed model satisfactorily describes integrated cellulose hydrolysis data taken with various reaction conditions (initial substrate concentration, initial product concentration, enzyme loading, time). Mechanistic (and semi-mechanistic) literature models for long-term enzymatic hydrolysis were compared with the modified HCH-1 model and evaluated by the corrected version of the Akaike information criterion. Comparison results show that the modified HCH-1 model provides the best fit for enzymatic cellulose hydrolysis. CONCLUSIONS: The HCH-1 model was modified to extend its application to integrated enzymatic hydrolysis; it performed well when predicting 10-day cellulose hydrolysis at various experimental conditions. Comparison with the literature models showed that the modified HCH-1 model provided the best fit.

Energy Portfolio Assessment Tool (EPAT): Sustainable energy planning using the WEF nexus approach - Texas case.

The paper introduces a holistic framework that identifies the links between energy and other systems (water, land, environment, finance, etc.), and measures the impact of energy portfolios, to offer a solid foundation for the best sustainable decision making in energy planning. The paper presents a scenario-based holistic nexus tool, Energy Portfolio Assessment Tool (EPAT) that provides a platform for energy stakeholders and policymakers to create and evaluate the sustainability of various scenarios based on the water-energy-food (WEF) nexus approach. The tool is applied to a case study in Texas, USA. Scenarios considered are set by the U.S. Energy Information Administration (EIA): EIA Reference Case - 2015, EIA Clean Power Plan (CPP) & Reference Case - 2030, and EIA No-CPP & Reference Case - 2030. In the presence of the CPP, total water withdrawal is expected to decrease significantly, while total water consumption is projected to experience a slight decrease due to the increase in water consumption in electricity generation caused by the new electricity mix. The CPP is successful in decreasing emissions, but is accompanied by tradeoffs, such as increased water consumption and land use by electricity generation. The absence of the CPP will lead to an extreme surge in total water withdrawn and consumed, and in emissions. Therefore, conservation policies should move from the silo to the nexus mentality to avoid unintended consequences that result in improving one part of the nexus while worsening the other parts.

Neural network prediction of biomass digestibility based on structural features.

Plots of biomass digestibility are linear with the natural logarithm of enzyme loading; the slope and intercept characterize biomass reactivity. The feed-forward back-propagation neural networks were performed to predict biomass digestibility by simulating the 1-, 6-, and 72-h slopes and intercepts of glucan, xylan, and total sugar hydrolyses of 147 poplar wood model samples with a variety of lignin contents, acetyl contents, and crystallinity indices. Regression analysis of the neural network models indicates that they performed satisfactorily. Increasing the dimensionality of the neural network input matrix allowed investigation of the influence glucan and xylan enzymatic hydrolyses have on each other. Glucan hydrolysis affected the last stage of xylan digestion, and xylan hydrolysis had no influence on glucan digestibility. This study has demonstrated that neural networks have good potential for predicting biomass digestibility over a wide range of enzyme loadings, thus providing the potential to design cost-effective pretreatment and saccharification processes.

Structural features affecting biomass enzymatic digestibility.

The rate and extent of enzymatic hydrolysis of lignocellulosic biomass highly depend on enzyme loadings, hydrolysis periods, and structural features resulting from pretreatments. Furthermore, the influence of one structural feature on biomass digestibility varies with the changes in enzyme loading, hydrolysis period and other structural features as well. In this paper, the effects of lignin content, acetyl content, and biomass crystallinity on the 1-, 6-, and 72-h digestibilities with various enzyme loadings were investigated. To eliminate the cross effects among structural features, selective pretreatment techniques were employed to vary one particular structural feature during a pretreatment, while the other two structural features remained unchanged. The digestibility results showed that lignin content and biomass crystallinity dominated digestibility whereas acetyl content had a lesser effect. Lignin removal greatly enhanced the ultimate hydrolysis extent. Crystallinity reduction, however, tremendously increased the initial hydrolysis rate and reduced the hydrolysis time or the amount of enzyme required to attain high digestibility. To some extent, the effects of structural features on digestibility were interrelated. At short hydrolysis periods, lignin content was not important to digestibility when crystallinity was low. Similarly, at long hydrolysis periods, crystallinity was not important to digestibility when lignin content was low.

Insights into Xylan Degradation and Haloalkaline Adaptation through Whole-Genome Analysis of Alkalitalea saponilacus, an Anaerobic Haloalkaliphilic Bacterium Capable of Secreting Novel Halostable Xylanase.

The obligately anaerobic haloalkaliphilic bacterium Alkalitalea saponilacus can use xylan as the sole carbon source and produce propionate as the main fermentation product. Using mixed carbon sources of 0.4% (w/v) sucrose and 0.1% (w/v) birch xylan, xylanase production from A. saponilacus was 3.2-fold greater than that of individual carbon sources of 0.5% (w/v) sucrose or 0.5% (w/v) birch xylan. The xylanse is halostable and exhibits optimal activity over a broad salt concentration (2⁻6% NaCl). Its activity increased approximately 1.16-fold by adding 0.2% (v/v) Tween 20. To understand the potential genetic mechanisms of xylan degradation and molecular adaptation to saline-alkali extremes, the complete genome sequence of A. saponilacus was performed with the pacBio single-molecule real-time (SMRT) and Illumina Misseq platforms. The genome contained one chromosome with a total size of 4,775,573 bps, and a G+C genomic content of 39.27%. Ten genes relating to the pathway for complete xylan degradation were systematically identified. Furthermore, various genes were predicted to be involved in isosmotic cytoplasm via the "compatible-solutes strategy" and cytoplasmic pH homeostasis though the "influx of hydrogen ions". The halostable xylanase from A. saponilacus and its genomic sequence information provide some insight for potential applications in industry under double extreme conditions.

Fixed-bed fermentation of rice straw and chicken manure using a mixed culture of marine mesophilic microorganisms.

A mixture of rice straw (80%) and chicken manure (20%) was pretreated and fermented to carboxylic acids by using a mixed culture of marine mesophilic microorganisms. Two sets of four fermentors, built from PVC pipes, were used for both biomass pretreatment and fermentation. Four 1L fermentors (F1-F4) were arranged in series, where liquid fermentation products were transferred from one fermentor to the other, to form a train. A liquid volume of 10mL and 15mL were transferred every four days for Trains A and B, respectively. The maximum total acid concentration for F1 in Train A was 34.2g/L and the maximum acid concentration in F2-F4 was approximately 44g/L. The maximum total acid concentration in F1 in Train B was 30.5g/L and the maximum acid concentration in F2-F4 was approximately 48g/L. The conversion in each of the fermentors in Train A varied from 0.821 to 0.879g VS digested/g VS fed and the yield was in the range 0.489-0.609g total acids/g VS fed. The conversion and yield in Train B were 0.741-0.914g VS digested/g VS fed and 0.563-0.669g total acids/g VS fed, respectively. The continuum particle distribution model (CPDM) predicted acid concentrations and retention times in the fixed-bed fermentation system with R(2) of 0.67-0.84 in Trains A and B.