Production of microbial oils co-fermented by Trichoderma.koningii and Trichosporon cutaneum with Daqu distiller's grains

Abstract： Daqu distiller＇s grains were co-fermented as raw materials by Trichoderma.koningii and Trichosporon cutaneum to produce microbial oils, which can provide raw materials for bio-diesel development. The single factor tests were used to investigate the effects of ratio of strains, inoculum size, culture temperature and culture time on the production of microbial oils. The best processing conditions were obtained by orthogonal test through measuring the content of microbial oils in product. The result shows that the microbial oils content of the co-fermented product under the ratio of Trichoderma.koningii to Trichosporon cutaneum of 1：1, the inoculum size of strains of 11% at 28 ℃ for 6 d is 7.15 g/L. It is shown that the production of microbial oils co-fermented by Trichoderma.koningii and Trichosporon cutaneum with Daqu distiller＇s grains is possible. The research provides a new idea for the reuse of Daqu distiller＇s grains and also provides a new way for the development of microbial oils.

Progress of Microbial Enhanced Oil Recovery in Laboratory Investigation

This paper describes a simple, easy process for screening microorganisms, and introduces a laboratory simulation device and process of microbial enhanced oil recovery (MEOR) , which is a necessary research step for trial in oilfields. The MEOR mechanism and the influence of adsorption, diffusion, metabolism, nutrition, porosity, and permeability are analyzed. The research indicates that different microbes have different efficiencies in EOR and that different culture types play different roles in EOR. The effect of syrup is better than that of glucose, and larger porosity is favorable to the reproduction and growth of microbes, thereby improving the oil recovery. Using crude oil as a single carbon source is more appreciable because of the decrease in cost of oil recovery. At the end of this paper, the development of polymerase chain reaction (PCR) for the future is discussed.

Research on the Jilin Oilfield Field Trials of Microbial Enhanced Oil Recovery

This paper describes the experience of Jilin oilfield trials for Microbial Enhanced Oil Recovery (MEOR). A new technique to identify microbes with DNA for MEOR has been established, and useful microbes selected for use in field trials. Behaviors of bacteria activated in the reservoir, oil recovery and water cut, and the viscosity of crude oil produced through huff & puff testing and flooding with molasses-injection tests, have been investigated in situ. CJF-002, which produces biopolysaccharide, is the best among the microbes used for field trials, as it can use molasses as nutrient and produce a small quantity of CO2 and a mass of water-insoluble biopolymer. The metabolic behavior in the reservoir showed that CJF-002 had a good potentiality for MEOR.

Study of a plugging microbial consortium using crude oil as sole carbon source

A microbial consortium named Y4 capable of producing biopolymers was isolated from petroleum-contaminated soil in the Dagang Oilfield, China. It includes four bacterial strains： Y4-1 （Paenibacillus sp.）, Y4-2 （Actinomadura sp.）, Y4-3 （Uncultured bacterium clone） and Y4-4 （Brevibacillus sp.）. The optimal conditions for the growth of the consortium Y4 were as follows： temperature about 46 ℃, pH about 7.0 and salinity about 20.0 g/L. The major metabolites were analyzed with gas chromatographymass spectrometry （GC-MS）. A comparison was made between individual strains and the microbial consortium for biopolymer production in different treatment processes. The experimental results showed that the microbial consortium Y4 could produce more biopolymers than individual strains, and the reason might be attributed to the synergetic action of strains. The biopolymers were observed with optical and electron microscopes and analyzed by paper chromatography. It was found that the biopolymers produced by the microbial consortium Y4 were insoluble in water and were of reticular structure, and it was concluded that the biopolymers were cellulose. Through a series of simulation experiments with sand cores, it was found that the microbial consortium Y4 could reduce the permeability of reservoir beds, and improve the efficiency of water flooding by growing biomass and producing biopolymers. The oil recovery was enhanced by 3.5% on average. The results indicated that the consortium Y4 could be used in microbial enhanced oil recovery and play an important role in bioremediation of oil polluted environments.

Relationship between Microbial Community Characteristics and Flooding Efficiency in Microbial Enhanced Oil Recovery

Microbial enhanced oil recovery (MEOR) is the research focus in the field of energy development as an environmentally friendly and low cost technology. MEOR can bes divided into indigenous microbial oil recovery and exogenous microbial oil recovery. The ultimate goal of indigenous microbial flooding is to enhance oil recovery via stimulation of specific indigenous microorganisms by injecting optimal nutrients. For studying the specific rule to activate the indigenous community during the long-term injection period, a series of indigenous displacement flooding experiments were carried out by using the long-core physical simulation test. The experimental results have shown that the movement of nutrients components (i.e., carbon/nitrogen/phosphorus) differed from the consumption of them. Moreover, there was a positive relationship between the nutrients concentration and bacteria concentration once observed in the produced fluid. And the trend of concentration of acetic acid was consistent with that of methanogens. When adding same activators, the impacts of selective activators to stimulate the indigenous microorganisms became worse along with the injection period, which led to less oil recovery efficiency.

Brief Review on MEOR Technology

As one of the main oil production countries, China pays great attention to the study and application of new technologies to enhance oil recovery. Microbial enhanced oil recovery (MEOR), an advanced technology, is one of the focuses. This paper gives a brief review on the history of the MEOR development and of laboratory-based and field application programmes. Mechanisms of MEOR are presented. Microorganisms can produce gases, acids, surfactants, polymers, all of which are capable of an effective improvement on the hydrocarbon structure, decreasing viscosity and also flooding oil physically under an increased pressure. Early field trials were initiated to utilize microorganisms to recover residual oil or gas. The microorganisms and their reaction products critically have an effect on oil and gas production in many reservoirs in China. Concerning research on metabolites, several field applications in some oil fields indicate that it is very significant to accurately analyse and evaluate metabolites and further select the most suitable microbe. Moreover, the MEOR program is applied in extra-heavy crude oilfields with relatively good results.

The Selection and Breeding of a Novel Microorganism Strain I and Investigation of Core Model Experiment for MEOR

This paper introduces the results of selecting and breeding a micro-organism, Strain I, and its core model experiment investigation for microbial enhanced oil recovery (MEOR). Strain I was separated from the formation water of the Dagang oil field, with analytical results showing that Strain I is a gram-positive bacillus. A further study revealed that this strain has an excellent tolerance of environmental stresses: It can survive in conditions of 70℃, 30 wt% salinity and pH3.5-9.4. Strain I can metabolize biosurfactants that could increase the oil recovery ratio, use crude oil as the single carbon source, and decompose long-chain paraffin with a large molecular weight into short-chain paraffin with a small molecular weight. The core model experiment shows that Strain I enhances oil recovery well. Using 2 vol% of the fermentation solution of Strain I to displace the crude oil in the synthetic plastic bonding core could increase the recovery ratio by 21.6%.

A Mutant Strain of a Surfactant-Producing Bacterium with Increased Emulsification Activity

As reported in this paper, a strain of oil-degrading bacterium Sp - 5 - 3 was determined to belong to Enterobacteriaceae, which would be useful for microbial enhanced oil recovery (MEOR). The aim of our study was to generate a mutant using low energy N+ beam implantation. With 10 keV of energy and 5.2× 1014 N+/cm2 of dose - the optimum condition, a mutant, S-34, was obtained, which had nearly a 5-fold higher surface and a 13-fold higher of emulsifica-tion activity than the wild type. The surface activity was measured by two methods, namely, a surface tension measuring instrument and a recording of the repulsive circle of the oil film; the emulsification activity was scaled through measuring the separating time of the oil-fermentation mixture. The metabolic acid was determined as methane by means of gas chromatography.

Laboratory studies of rice bran as a carbon source to stimulate indigenous microorganisms in oil reservoirs

There is a great interest in developing cost-efficient nutrients to stimulate microorganisms in indigenous microbial enhanced oil recovery（IMEOR） processes.In the present study,the potential of rice bran as a carbon source for promoting IMEOR was investigated on a laboratory scale.The co-applications of rice bran,K2HPO4 and urea under optimized bio-stimulation conditions significantly increased the production of gases,acids and emulsifiers.The structure and diversity of microbial community greatly changed during the IMEOR process,in which Clostridium sp.,Acidobacteria sp.,Bacillus sp.,and Pseudomonas sp.were dominant.Pressurization,acidification and emulsification due to microbial activities and interactions markedly improved the IMEOR processes.This study indicated that rice bran is a potential carbon source for IMEOR.

CHARACTERISTICS OF MICROBIAL TRANSFUSION IN POROUS MEDIUM AND ITS THEORETICAL INVESTIGATION

This paper introduces the experimental and theoretical results of microbialtransfusion in porous medium. The experimetnal results show that microbes can pass through throatsof porous media which permeability is 0. 03 Darcy or over. Based on these results a new supplemorphotropy and dynamic directionality theory is obtained to explain this transfusion ability.Considering the characteristics of convection and diffusion while microbe is moving, one dimensionaland radial transfusion mathematical model including adsorption has been set up. With this model andexperimental data, the diffusion coefficient and maximum adsorption constants are obtained bycalculation and mathematical fitting. The results show that the concentration of microbe decreasesquickly during transfusion, and the effective region also decreases. Therefore, it is indispensablefor microbes to reproduce quickly under the reservoir conditions. Additionally, this model can alsobe used to calculate the time when the oil production begins to increase and when the microbes aregot from another production well.

Application of thermotolerant petroleum microbes at reservoir conditions for enhanced oil recovery

Primary oil recovery is the first stage of hydrocarbon production in which a reservoir uses its natural energy to force hydrocarbon to its wellbore.Secondary oil recovery comes to play when hydrocarbons can no longer be further produced by natural means.The purpose of secondary recovery is to maintain reservoir pressure so as to displace hydrocarbons toward the wellbore.Both primary and secondary recovery processes cannot displace more than 50%of the available hydrocarbons in a reservoir.The remaining hydrocarbons are further recovered through Tertiary/Enhanced Oil Recovery techniques.According to literature,microbial enhanced oil recovery has been identified as a tertiary method used to improve the efficiency of hydrocarbon production from reservoirs.Microbial enhanced oil recovery is a feasible reservoir technology,which has not been widely used in the oil and gas industry owing to the attainment of the requisite reservoir conditions such as temperature within which microbes can thrive.Literature has shown that thermotolerant microbes can withstand optimum temperatures of 50e90℃,while deep and ultra-deep hydrocarbon reservoir temperatures are often above 100℃.This study identifies some isolated thermotolerant microbes from a sandstone reservoir that can withstand temperatures as high as 110℃via conventional methods and molecular analysis.The identified thermotolerant petroleum microbes:Bacillus amyloliquefaciens(A)and Bacillus nealsonii(B)were used to enhance oil recovery from a reservoir.The results showed that the microbial species A and B at a confined pressure of 3.0 MPa and temperature of 27℃,gave 46.4%and 48.6%oil recoveries,respectively,which is comparably higher than the value(26.9%)obtained for the water flooded samples.At temperatures of 80,90,100,110 and 120℃,the oil recovery results show that the recovery factor(55.2%e64.1%)of species B were higher compared to the range(46.7e57.5%)recorded for species A.At the onset of the core flooding experiments,there was an initial increment in oil recovery factor as the temperature increased from 80 to 110℃,whereas,it remained constant within 110e120℃.This trend coincides with the drop in the thermal resistance exhibited by the microbes when exposed to such conditions.The cumulative oil production from the commercial Eclipse simulation closely matched those of the experiment results,whereas,the slight difference can be attributed to the adjustment of the simulation input parameters.The experimental results show that species B can be used to enhance oil recovery at reservoir temperature conditions above 100℃.

Role of biosurfactants in bioremediation of oil pollution-a review

The energy resources mainly petroleum and petroleum hydrocarbons are major pollutants of the environment.The oil and oil products contamination may cause severe harm and hence,the attention has been remunerated in the development of alternative technologies for elimination of these contaminants.Biosurfactants were used in the remediation of oil pollution due to advantages such as biodegradability and low toxicity.The biosurfactants are produced from low cost substrates like agro-industrial wastes which reduce the cost of production.Biosurfactants and bioemulsifiers are amphiphilic compounds and are produced as extracellular or a part of the cell membrane by bacteria.The insight view,how hydrocarbons are degraded by microorganisms and thereby reduce the damage of ecosystem is highly essential to target the problem.Biofilms are the bacterial communities which protects the bacterial cells from various adverse conditions.The present review describes the biosurfactants and its synthesis from bacteria and also emphases on the role of surfactants in oil remediation.