Research on the Nature of Thermophilic Anaerobic Ethanol Producer Thermo anaerobacter sp DF3 in Petroleum Reservoirs

[Objective] The aim was to study the physiological and biochemical char- acteristics of Thermoanaerobacter sp DF3 in petroleum reservoirs and optimize the culture plan of producing ethanol from xylose. [Method] DF3, an anaerobic bacillus producing ethanol, was isolated from produced liquid from oil layer of Dagang oil field with anaerobic isolation technique. The phylogenetic position was analyzed by physiological and biochemical identification and phylogeny of 16S rDNA sequence. The metabolites were analyzed by gas chromatograph. [Result] The strain DF3 was a strict anaerobic thermophilic bacterium, which was straight in rod shape,and gram negative. Besides, it was 0.42 μmx（1.60-5.20） iJm in length. The strains can be soli- tary,in pairs or string and apical spore usually produced. Its growth temperature was 45-78 and 65 ℃ was the optimum. Many substances could be used as carbon sources, including glucose, xylose, fructose, ribose, mannose, arabinose, sucrose, galactose, lactose, cellobiose, melizitose, raffinose-, and starch. The similarity between strain DF3 and T. pseudoethanolicus achieved 99.7%. The main product of glucose and xylose fermentation was ethanol. After the culture plan was optimized,the final concentration of ethanol was 2.0 g/L. [Conclusion] It was proved through experiments that the strain DF3 was one of the strains with higher activity to produce ethanol at present and it could produce 2.0 g/L ethanol from xylose metabolization at 65 ℃. It has been demonstrated that DF3 is one of the known strains with high-production to produce ethanol,for example, 2.0 g/L ethano at 65℃. Presently, all the high-yield ethanol can be produced from metabolic xylose strains of metabolic xylose were isolat- ed by foreign countries, therefore, isolation of strain DF3 has provided an excellent original strain for studying ethanol production from lignocellulose in China.

CHARACTERISTICS OF THERMOPHILIC ANAEROBIC ACIDOGENESIS OF STARCH WASTEWATER

Acidogenic dissimilation of synthetic starch wastewater (1 000～10 000 mg COD·L -1 ) was studied in a thermophilic (55 ℃) upflow anaerobic sludge blanket (UASB) reactor.The production of volatile fatty acids (VFA) was proportional to the chemical oxygen demand (COD) loading rate.The yield of VFA was around 0.28 g VFA/g COD over the COD loading rate from 1.25 to 30 g COD·L -1 ·d -1 and the hydraulic retention time from 8.8 h to 24 h.Distribution of organic acids,the contents of propionic and butyric acids in the effluent in particular were also dependent on the COD loading rate.The thermophilic UASB reactor showed a stable performance on hydrolysis and acidogenesis of starch as well as suspended solid removal at short hydraulic retention times and high influent pH(10～11),during the operation of 110 d.

Influence of temperature fluctuation on thermophilic anaerobic digestion of municipal organic solid waste

A laboratory-scale experiment was carried out to assess the influence of temperature fluctuation on thermophilic anaerobic digestion of municipal organic solid waste （MOSW）. Heating failure was simulated by decreasing temperature suddenly from 55 ℃ to 20 ℃ suddenly, 2 h time is needed for temperature decrease and recovery. Under the conditions of 8.0 g/（L·d） and 15 d respectively for MOSW load and retention time, following results were noted： （1） biogas production almost stopped and VFA （volatile fatty acid） accumulated rapidly, accompanied by pH decrease; （2） with low temperature （20 ℃） duration of 1, 5, 12 and 24 h, it took 3, 11, 56 and 72 h for the thermophilic anaerobic digestion system to reproduce methane after temperature fluctuation, （3） the longer the low temperature interval lasted, the more the methanogenic bacteria would decay, hydrolysis, acidification and methanogenesis were all influenced by temperature fluctuation： （4） the thermophilic microorganisms were highly resilient to temperature fluctuation.

Enhancement of thermophilic anaerobic digestion of thickened waste activated sludge by combined microwave and alkaline pretreatment

Pretreatment of thickened waste activated sludge （TWAS） by combined microwave and alkaline pretreatment （MAP） was studied to improve thermophilic anaerobic digestion efficiency. Uniform design was applied to determine the combination of target temperature （110-210°C）, microwave holding time （1-51 min）, and NaOH dose （0-2.5 g NaOH/g suspended solids （SS）） in terms of their effect on volatile suspended solids （VSS） solubilization. Maximum solubilization ratio （85.1%） of VSS was observed at 210°C with 0.2 g-NaOH/g-SS and 35 min holding time. The effects of 12 different pretreatment methods were investigated in 28 thermophilic batch reactors by monitoring cumulative methane production （CMP）. Improvements in methane production in the TWAS were directly related to the microwave and alkaline pretreatment of the sludge. The highest CMP was a 27% improvement over the control. In spite of the increase in soluble chemical oxygen demand concentration and the decrease in dewaterability of digested sludge, a semi-continuous thermophilic reactor fed with pretreated TWAS without neutralization （at 170~C with 1 min holding time and 0.05 g NaOH/g SS） was stable and functioned well, with volatile solid （VS） and total chemical oxygen demand （TCOD） reductions of 28% and 18%, respectively, which were higher than those of the control system. Additionally, methane yields （L@STP/g-CODaded, at standard temperature and pressure （STP） conditions of 0°C and 101.325 kPa） and （L@STP/g VSadad） increased by 17% and 13%, respectively, comoared to the control reactor.

Degradation of polyacrylamide (PAM) and methane production by mesophilic and thermophilic anaerobic digestion: Effect of temperature and concentration

Polyacrylamide(PAM)is generally employed in wastewater treatment processes such as sludge dewatering and therefore exists in the sludge.Furthermore,it degrades slowly and can deteriorate methane yield during anaerobic digestion(AD).The impact or fate of PAM in AD under thermophilic conditions is still unclear.This study mainly focuses on PAM degradation and enhanced methane production from PAM-added sludge during 15 days of thermophilic(55°C)AD compared to mesophilic(35°C)AD.Sludge and PAM dose from 10 to 50 g/kg TSS were used.The results showed that PAM degraded by 76%to 78%with acrylamide(AM)content of 0.2 to 3.3 mg/L in thermophilic AD.However,it degraded only 21%to 30%with AM content of 0.5 to 7.2 mg/L in mesophilic AD.The methane yield was almost 230 to 238.4 mL/g VSS on the 8th day in thermophilic AD but was 115.2 to 128.6 mL/g VSS in mesophilic AD.Mechanism investigation revealed that thermophilic AD with continuous stirring not only enhanced PAM degradation but also boosted the organics release from the sludge with added PAM and gave higher methane yield than mesophilic AD.

Response of anaerobes to methyl fluoride,2-bromoethanesulfonate and hydrogen during acetate degradation

To use the selective inhibition method for quantitative analysis of acetate metabolism in methanogenic systems,the responses of microbial communities and metabolic activities,which were involved in anaerobic degradation of acetate,to the addition of methyl fluoride（CH3F）,2-bromoethanesulfonate（BES）and hydrogen were investigated in a thermophilic batch experiment.Both the methanogenic inhibitors,i.e.,CH3F and BES,showed their effectiveness on inhibiting CH4 production,whereas acetate metabolism other than acetoclastic methanogenesis was stimulated by BES,as reflected by the fluctuated acetate concentration.Syntrophic acetate oxidation was thermodynamically blocked by hydrogen（H2）,while H2-utilizing reactions as hydrogenotrophic methanogenesis and homoacetogenesis were correspondingly promoted.Results of PCR-DGGE fingerprinting showed that,CH3F did not influence the microbial populations significantly.However,the BES and hydrogen notably altered the bacterial community structures and increased the diversity.BES gradually changed the methanogenic community structure by affecting the existence of different populations to different levels,whilst H2 greatly changed the abundance of different methanogenic populations,and induced growth of new species.