Methane Potential and Microbial Community Dynamics in Anaerobic Digestion of Silage and Dry Cornstalks: a Substrate Exchange Study

Silage and dry are the two typical cornstalk forms. Either form could be used as substrate in biogas plants and might be replaced by another when shortage occurred. This study focused on the feeding sequence of these two kinds of feedstocks, aiming to discuss their specific methane potential (SMP). A 15-day hydraulic retention time was chosen for semi-continuous experiments based on the batch test results. In semi-continuous experiments, before and after feedstocks were exchanged, the significantly decreased and comparable SMPs of silage and dry cornstalks indicated that a basis of unstable digestion would result in incomplete methane release from the subsequent digestion. A higher similarity of bacterial community structure and greater quantity of bacteria were shown in acidified silage cornstalk digestion through band similarity analysis. Methanosaetaceae and methanomicrobiales were the predominant methanogens, and aceticlastic methanogenesis was the main route for methane production. The different feeding sequences affected the hydrolysis course and further influenced the methanogenic proliferation. Our work suggests that silage cornstalk digestion should be conducted before dry cornstalk digestion.     

Analysis of Relationship Between Microbial and Methanogenic Biomass in Methane Fermentation

To analyze the relationship between biomass of microorganisms and methane production, the total biomass of bacteria and archaea (BA) during methane fermentation was analyzed by the environmental DNA analysis method. In the case of using methanogenic sludge as a seed which is generally used for methane fermentation, the total BA biomass reached to 1.5 × 10⁸ to 3.6 × 10⁸ cells/ml when methane was produced. On the other hand, soil suspension was used as a seed; methane was not produced for 14-day cultivation. However, the total BA biomass reached to above 1.5 × 10⁸ cells/ml. The methanogen biomass was counted by using a fluorescence microscope (coenzyme F₄₂₀), and the methanogen biomass and the ratio of methanogens in the total of BA were analyzed during methane fermentation. At the methane-producing phase, the methanogen biomass reached to 1.3 × 10⁸ cells/ml, and the ratio of methanogens was above 70% of the total BA. When the ratio of methanogens in a seed was changed, the methane-producing phase was moved. However, the relationship between methanogens and other microorganisms at the methane-producing phase was almost similar.     

质量混合比对餐厨垃圾厌氧发酵产氢的影响

采用自制的序批式"厌氧发酵产氢反应装置",以餐厨垃圾为发酵底物,污水处理厂剩余污泥为接种微生物,初始pH为7.0、温度为37℃条件下,研究餐厨垃圾和剩余污泥不同质量混合比对厌氧发酵产氢的影响。结果表明,餐厨垃圾和剩余污泥质量比为4∶1时,厌氧发酵产氢效果最好,累积产氢量和单位产氢量(以VS计)最大,分别为252.2 m L和53.3 m L/g,TS和VS的去除率分别为20.9%和13.8%。     

Toward Plant Cell Wall Degradation Under Thermophilic Condition: A Unique Microbial Community Developed Originally from Swine Waste

A unique thermophilic microbial community developed initially from swine waste was investigated in this study. Cellulase activities were observed when this community was inoculated to media containing either cellulose or carboxymethylcellulose at 57 °C. Through constructing a clone library for the 16S ribosomal DNA, it was revealed that this community was mainly composed of three genera: Thermobacillus, Brevibacillus, and Anoxybacillus. New findings regarding the thermo- and pH stability of crude cellulases secreted by Brevibacillus sp. JXL were presented. Recent study on the growth characteristics of Anoxybacillus sp. 527 was discussed.     

Utilization of Waste Biomass (Kitchen Waste) Hydrolysis Residue as Adsorbent for Dye Removal: Kinetic,Equilibrium, and Thermodynamic Studies

Kitchen waste hydrolysis residue (KWHR), which is produced in the bioproduction process from kitchen waste (KW), is usually wasted with potential threats to the environment. Herein, experiments were carried out to evaluate the potential of KWHR as adsorbent for dye (methylene blue, MB) removal from aqueous solution. The adsorbent was characterized using FT-IR and SEM. Adsorption results showed that the operating variables had great effects on the removal efficiency of MB. Kinetic study indicated pseudo-second-order model was suitable to describe the adsorption process. Afterwards, the equilibrium data were well fitted by using Langmuir isotherm model, suggesting a monolayer adsorption. The Langmuir monolayer adsorption capacity was calculated to be 110.13 mg/g, a level comparable to some other low-cost adsorbents. It was found that the adsorption process of MB onto KWHR was spontaneous and exothermic through the estimation of thermodynamic parameters. Thus, KWHR was of great potential to be an alternative adsorbent material to improve the utilization efficiency of bioresource (KW) and lower the cost of adsorbent for color treatment.     

Parameter Estimation for Simultaneous Saccharification and Fermentation of Food Waste Into Ethanol Using Matlab Simulink

The increase in waste disposal and energy costs has provided an incentive to convert carbohydrate-rich food waste streams into fuel. For example, dining halls and restaurants discard foods that require tipping fees for removal. An effective use of food waste may be the enzymatic hydrolysis of the waste to simple sugars and fermentation of the sugars to ethanol. As these wastes have complex compositions which may change day-to-day, experiments were carried out to test fermentability of two different types of food waste at 27 degrees C using Saccharomyces cerevisiae yeast (ATCC4124) and Genencor's STARGEN enzyme in batch simultaneous saccharification and fermentation (SSF) experiments. A mathematical model of SSF based on experimentally matched rate equations for enzyme hydrolysis and yeast fermentation was developed in Matlab Simulink. Using Simulink parameter estimation 1.1.3, parameters for hydrolysis and fermentation were estimated through modified Michaelis-Menten and Monod-type equations with the aim of predicting changes in the levels of ethanol and glycerol from different initial concentrations of glucose, fructose, maltose, and starch. The model predictions and experimental observations agree reasonably well for the two food waste streams and a third validation dataset. The approach of using Simulink as a dynamic visual model for SSF represents a simple method which can be applied to a variety of biological pathways and may be very useful for systems approaches in metabolic engineering in the future.     

Enzymatic Hydrolysis and Ethanol Fermentation of High Dry Matter Wet-Exploded Wheat Straw at Low Enzyme Loading

Wheat straw was pretreated by wet explosion using three different oxidizing agents (H₂O₂, O₂, and air). The effect of the pretreatment was evaluated based on glucose and xylose liberated during enzymatic hydrolysis. The results showed that pretreatment with the use of O₂ as oxidizing agent was the most efficient in enhancing overall convertibility of the raw material to sugars and minimizing generation of furfural as a by-product. For scale-up of the process, high dry matter (DM) concentrations of 15–20% will be necessary. However, high DM hydrolysis and fermentation are limited by high viscosity of the material, higher inhibition of the enzymes, and fermenting microorganism. The wet-explosion pretreatment method enabled relatively high yields from both enzymatic hydrolysis and simultaneous saccharification and fermentation (SSF) to be obtained when performed on unwashed slurry with 14% DM and a low enzyme loading of 10 FPU/g cellulose in an industrial acceptable time frame of 96 h. Cellulose and hemicellulose conversion from enzymatic hydrolysis were 70 and 68%, respectively, and an overall ethanol yield from SSF was 68%.     

Industrial Waste Utilization for Low-Cost Production of Raw Material Oil Through Microbial Fermentation

In view of ever-growing demand of biodiesel, there is an urgent need to look for inexpensive and promising renewable raw material oils for its production. In this context, the aim of this study was to evaluate the potential use of industrial wastes for low-cost production of oils through microbial fermentation. Among the strains tested, Yarrowia lipolytica grew best and produced highest lipid when grown on decanter effluent from palm oil mill. When crude glycerol by-product from a biodiesel plant was added into the effluent as a co-substrate, Y. lipolytica produced a higher biomass of 3.21 g/L and a higher amount of lipid of 2.21 g/L which was 68 % of the dry weight. The scale up and process improvement in a 5-L bioreactor increased the biomass and lipid up to 5.53 and 2.81 g/L, respectively. A semi-continuous mode of operation was an effective mode for biomass enhancement while a fed-batch mode was effective for lipid enhancement. These yeast lipids have potential to be used as biodiesel feedstocks because of their similar fatty acid composition to that of plant oil.     

Production of Ethanol and Feed by High Dry Matter Hydrolysis and Fermentation of Palm Kernel Press Cake

Palm kernel press cake (PKC) is a residue from palm oil extraction presently only used as a low protein feed supplement. PKC contains 50% fermentable hexose sugars present in the form of glucan and mainly galactomannan. This makes PKC an interesting feedstock for processing into bioethanol or in other biorefinery processes. Using a combination of mannanase, β-mannosidase, and cellulases, it was possible without any pretreatment to hydrolyze PKC at solid concentrations of 35% dry matter with mannose yields up to 88% of theoretical. Fermentation was tested using Saccharomyces cerevisiae in both a separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) setup. The hydrolysates could readily be fermented without addition of nutrients and with average fermentation yields of 0.43?±?0.02 g/g based on consumed mannose and glucose. Employing SSF, final ethanol concentrations of 70 g/kg was achieved in 216 h, corresponding to an ethanol yield of 70% of theoretical or 200 g ethanol/kg PKC. Testing various enzyme mixtures revealed that including cellulases in combination with mannanases significantly improved ethanol yields. Processing PKC to ethanol resulted in a solid residue enriched in protein from 17% to 28%, a 70% increase, thereby potentially making a high-protein containing feed supplement.     

Thermophilic Bacillus coagulans Requires Less Cellulases for Simultaneous Saccharification and Fermentation of Cellulose to Products than Mesophilic Microbial Biocatalysts

Ethanol production from lignocellulosic biomass depends on simultaneous saccharification of cellulose to glucose by fungal cellulases and fermentation of glucose to ethanol by microbial biocatalysts (SSF). The cost of cellulase enzymes represents a significant challenge for the commercial conversion of lignocellulosic biomass into renewable chemicals such as ethanol and monomers for plastics. The cellulase concentration for optimum SSF of crystalline cellulose with fungal enzymes and a moderate thermophile, Bacillus coagulans, was determined to be about 7.5 FPU g^?1 cellulose. This is about three times lower than the amount of cellulase required for SSF with Saccharomyces cerevisiae, Zymomonas mobilis, or Lactococcus lactis subsp. lactis whose growth and fermentation temperature optimum is significantly lower than that of the fungal cellulase activity. In addition, B. coagulans also converted about 80% of the theoretical yield of products from 40 g/L of crystalline cellulose in about 48 h of SSF with 10 FPU g^?1 cellulose while yeast, during the same period, only produced about 50% of the highest yield produced at end of 7 days of SSF. These results show that a match in the temperature optima for cellulase activity and fermentation is essential for decreasing the cost of cellulase in cellulosic ethanol production.     

Bioethanol Production from Soybean Residue via Separate Hydrolysis and Fermentation

Bioethanol was produced using polysaccharide from soybean residue as biomass by separate hydrolysis and fermentation (SHF). This study focused on pretreatment, enzyme saccharification, and fermentation. Pretreatment to obtain monosaccharide was carried out with 20% (w/v) soybean residue slurry and 270 mmol/L H₂SO₄ at 121 °C for 60 min. More monosaccharide was obtained from enzymatic hydrolysis with a 16 U/mL mixture of commercial enzymes C-Tec 2 and Viscozyme L at 45 °C for 48 h. Ethanol fermentation with 20% (w/v) soybean residue hydrolysate was performed using wild-type and Saccharomyces cerevisiae KCCM 1129 adapted to high concentrations of galactose, using a flask and 5-L fermenter. When the wild type of S. cerevisiae was used, an ethanol production of 20.8 g/L with an ethanol yield of 0.31 g/g consumed glucose was obtained. Ethanol productions of 33.9 and 31.6 g/L with ethanol yield of 0.49 g/g consumed glucose and 0.47 g/g consumed glucose were obtained in a flask and a 5-L fermenter, respectively, using S. cerevisiae adapted to a high concentration of galactose. Therefore, adapted S. cerevisiae to galactose could enhance the overall ethanol fermentation yields compared to the wild-type one.     

Bioconversion of Welan Gum from Kitchen Waste by a Two-Step Enzymatic Hydrolysis Pretreatment

Kitchen waste (KW) is a worldwide issue, which can lead to environment pollution. Nevertheless, it is also a low-cost and sustainable resource for bio-production. Meanwhile, welan gum (WG) is one kind of the most important exopolysaccharide but with high material cost. The aim of this study was to adopt two-step enzymatic hydrolysis to improve the release and recovery of both sugar and protein in KW for subsequent WG production. As the results, the recovery rates of sugar and protein reached 81.07 and 77.38%, which were both satisfactory. After the conditions optimized in flasks, the welan fermentation was conducted in a 5-L fermentor, and the WG yield, utilization rates of reducing sugar and KDN, respectively, reached 5.57 g L^?1, 94.25% and 61.96%. Moreover, the kinetic analyses demonstrated that the WG fermentation in KWH was a partly growth-associated process. The KW was successfully treated by fermentation for the bioconversion to WG.     

乙醇微生物电极的研制及应用

用ＰＡＶ、纤维素等材料将从土壤分离得到的一株好氧乙醇菌包埋制，备的微生物膜与极谱型氧电极结合，制成乙醇微生物电极，其线性范围为０．３－１２．０ｍｇ／Ｌ，响应时间小于５ｍｉｎ，测定的最适宜ＰＨ为６．４－７．３，电极连续全用三个月信号基本稳定。     

Extraction of Astaxanthin from Shrimp Waste Using Pressurized Hot Ethanol

An efficient and environmentally sustainable extraction method is proposed for the enrichment of a high-value pigment, astaxanthin, from a low-value raw material, shrimp waste. Ethanol at elevated temperature and pressure was used as a “green” extraction solvent. An experimental design approach based on central composite design was used to investigate the dependence of pressurized liquid extraction (PLE) operating variables (pressure, temperature, extraction time) on the recovered astaxanthin concentration from shrimp waste. The results show that at a 95% confidence level, the most significant PLE operating variables were extraction temperature and time. Extraction pressure had only a minor effect on the astaxanthin recovery in the studied experimental conditions. The maximum astaxanthin recovery obtainable by PLE was calculated from the chemometrics results and then appraised by experiments. Our results show astaxanthin yields of around 24 mg kg^?1 shrimp waste. The reproducibility of the developed PLE method is good, showing a relative standard deviation of 3.5% (n = 6) for astaxanthin.     

The Operable Modeling of Simultaneous Saccharification and Fermentation of Ethanol Production from Cellulose

An operable batch model of simultaneous saccharification and fermentation (SSF) for ethanol production from cellulose has been developed. The model includes four ordinary differential equations that describe the changes of cellobiose, glucose, yeast, and ethanol concentrations with respect to time. These equations were used to simulate the experimental data of the four main components in the SSF process of ethanol production from microcrystalline cellulose (Avicel PH101). The model parameters at 95% confidence intervals were determined by a MATLAB program based on the batch experimental data of the SSF. Both experimental data and model simulations showed that the cell growth was the rate-controlling step at the initial period in a series of reactions of cellulose to ethanol, and later, the conversion of cellulose to cellobiose controlled the process. The batch model was extended to the continuous and fed-batch operating models. For the continuous operation in the SSF, the ethanol productivities increased with increasing dilution rate, until a maximum value was attained, and rapidly decreased as the dilution rate approached the washout point. The model also predicted a relatively high ethanol mass for the fed-batch operation than the batch operation.     

甲烷干重整反应用Ni-Ru/MgAl类水滑石催化剂的研究

采用焙烧记忆法分别制备Ni/Mg Al O和NiRu/Mg Al O类水滑石催化剂用于甲烷干重整反应.利用XRD、TPR、TG、XPS、CO2-TPD、TEM等表征催化剂的结构及失活特征,发现在Ni/Mg Al O中添加Ru,有利于增加催化剂表面Ni含量,并促进Ni2+的还原.不同Mg/Al比双金属催化剂中,7Ni-0.15Ru/Mg2.5Al催化剂具有较高的催化活性,这归结为该催化剂适宜的碱性、较高表面Ni含量以及小尺寸的Ni0物种.添加Ru明显抑制Ni/Mg Al O催化剂表面的丝状碳的形成.而7Ni-0.15Ru/Mg2.5Al较强的抗积碳性能与其较小Ni0晶粒尺寸及适宜催化剂碱性有关.     

Detoxification and Fermentation of Pyrolytic Sugar for Ethanol Production

The sugars present in bio-oil produced by fast pyrolysis can potentially be fermented by microbial organisms to produce cellulosic ethanol. This study shows the potential for microbial digestion of the aqueous fraction of bio-oil in an enrichment medium to consume glucose and produce ethanol. In addition to glucose, inhibitors such as furans and phenols are present in the bio-oil. A pure glucose enrichment medium of 20?g/l was used as a standard to compare with glucose and aqueous fraction mixtures for digestion. Thirty percent by volume of aqueous fraction in media was the maximum additive amount that could be consumed and converted to ethanol. Inhibitors were removed by extraction, activated carbon, air stripping, and microbial methods. After economic analysis, the cost of ethanol using an inexpensive fermentation medium in a large scale plant is approximately $14 per gallon.     

分子对接和动力学模拟提高嗜热蛋白酶PhpI的活力

通过分子对接和动力学模拟对嗜热蛋白酶的分子进行改造, 确定蛋白酶PH1704(PhpI)定点突变残基, 并通过分子生物学实验进行验证. 突变体K43C的蛋白酶活力提高了5.8倍. 分子动力学模拟结果表明, 经过8 ns的动力学模拟后, K43C突变体二级结构由野生型的S2片层(F11-E12-D13)变成环状结构. E12和K43均是活性位点的重要残基, 这种变化将导致活性位点的柔性增强, 有利于催化反应的发生.     

Optimized Fed-Batch Fermentation of Scheffersomyces stipitis for Efficient Production of Ethanol from Hexoses and Pentoses

Scheffersomyces stipitis was cultivated in an optimized, controlled fed-batch fermentation for production of ethanol from glucose–xylose mixture. Effect of feed medium composition was investigated on sugar utilization and ethanol production. Studying influence of specific cell growth rate on ethanol fermentation performance showed the carbon flow towards ethanol synthesis decreased with increasing cell growth rate. The optimum specific growth rate to achieve efficient ethanol production performance from a glucose-xylose mixture existed at 0.1 h^?1. With these optimized feed medium and cell growth rate, a kinetic model has been utilized to avoid overflow metabolism as well as to ensure a balanced feeding of nutrient substrate in fed-batch system. Fed-batch culture with feeding profile designed based on the model resulted in high titer, yield, and productivity of ethanol compared with batch cultures. The maximal ethanol concentration was 40.7 g/L. The yield and productivity of ethanol production in the optimized fed-batch culture was 1.3 and 2 times higher than those in batch culture. Thus, higher efficiency ethanol production was achieved in this study through fed-batch process optimization. This strategy may contribute to an improvement of ethanol fermentation from lignocellulosic biomass by S. stipitis on the industrial scale.     

Fermentation of Sugarcane Bagasse and Chicken Manure to Calcium Carboxylates under Thermophilic Conditions

Sugarcane bagasse and chicken manure were anaerobically fermented to carboxylic acids using a mixed culture of marine microorganisms at 55 °C. Using the MixAlco process— an example of consolidated bioprocessing— the resulting carboxylate salts can be converted to mixed alcohol fuels or gasoline. To enhance digestibility, sugarcane bagasse was lime pretreated with 0.1 g Ca(OH)₂/g dry biomass at 100 °C for 2 h. Four-stage countercurrent fermentation of 80% sugarcane bagasse/20% chicken manure was performed at various volatile solids (VS) loading rates and liquid residence times. Calcium carbonate was used as a buffer during fermentation. The highest acid productivity of 0.79 g/(L day) occurred at a total acid concentration of 21.5 g/L. The highest conversion (0.59 g VS digested/g VS fed) and yield (0.18 g total acids/g VS fed) occurred at a total acid concentration of 15.5 g/L. The continuum particle distribution model (CPDM) predicted the experimental total acid concentrations and conversions at an average error of 10.14% and 12.68%, respectively. CPDM optimizations show that high conversion (>80%) and total acid concentration of 21.3 g/L are possible with 300 g substrate/(L liquid), 30 days liquid residence time, and 3 g/(L day) solid loading rate. Thermophilic fermentation has a higher acetate content (∼63 wt%) than mesophilic fermentation (∼39 wt%).