Bolaform lipid composition, production method and use thereof

Abstract

The present invention discloses a biotechnological method to produce a Thermotoga bolaform lipid (BL) composition, and shows the benefits of the use thereof in enhanced hydrocarbon de-emulsification and recovery processes. The LB composition has viscoelastic, de-emulsifying, dispersing, wettability-modifying, surface tension and crude oil viscosity reducing properties. The composition demonstrates stability, compatibility and functionality in carbonate reservoirs, sandstones, siltstones, shales or in heterogeneous lithologies, as well as under high temperature conditions, high divalent ion content, high salinity or pH variations. The invention induces the formation of a viscoelastic interphase that separates the aqueous phase from the oily phase, which overcomes the flow of fluids through preferential paths, which breaks down emulsions and significantly improves oil sweeping and desorption at concentrations as low as 0.005 to 0.01% by weight.

Description

[0001] COMPOSITION OF BOLAFORM LIPIDS; METHOD OF OBTAINING THEM; AND THEIR USE

[0002] TECHNICAL FIELD OF THE INVENTION

[0003] This invention relates to bioprocesses for the biosynthesis and production of a viscoelastic surfactant composition from Thermotoga' bolaform lipids, as well as to the benefit of their application, at ultra-low concentrations, in demulsification and enhanced hydrocarbon recovery processes. The bolaform lipids demonstrate stability, compatibility, and functionality in carbonate reservoirs, sandstones, mudstones, shales, or in heterogeneous lithologies, as well as under conditions of high temperature, high divalent ion content, high salinity, or pH variations. The bolaform lipid composition possesses viscoelastic, demulsifying, dispersing, wettability-modifying, viscosity-reducing, and interfacial tension-reducing properties.It induces the formation of a viscoelastic interface that separates the aqueous phase from the oily phase, which manages to overcome the flow of fluids through preferential paths, which breaks emulsions and which improves the scavenging and desorption of oil.

[0004] BACKGROUND OF THE INVENTION

[0005] The production of fossil fuels from crude oil presents two main technological challenges: (i) removing and separating emulsified water from the oil, and (ii) forcing the reservoir to deliver the maximum original in-situ volume of crude oil. The challenge, therefore, is to develop and implement efficient, cost-effective, and environmentally friendly demulsification and enhanced oil recovery (EOR) technologies and processes.

[0006] The objective of enhanced oil recovery is to decrease oil saturation below residual oil saturation (RES) once the primary and secondary recovery stages have been exhausted, when production becomes discontinuous and reaches its economic limit.

[0007] Reservoirs tend to retain oil due to physicochemical properties that affect fluid flow in porous media. For example, the preferential wettability of rock to oil over water; high capillary pressures, high and low permeability zones; high viscosity; and high amounts of asphaltenes in the crude oil, among others. Recovering the oil retained in the formation means extending the useful life of the reservoir by forcing it to deliver the maximum value of the original in-situ volume of crude oil.

[0008] A recurring technology for EOR is surfactant flooding with the purpose of modifying the physicochemical properties of the fluid flow and / or the rock through the following main mechanisms: i) increasing the displacement mobility, increasing the viscosity of the injection water or decreasing that of the oil or both; i) reducing the interfacial tension between immiscible fluids, i¡¡) modifying the wettability of the rock formation from oil-wet to water-wet, iv) decreasing the interfacial tension and v) favoring the fluid flow causing the formation of a viscoelastic interface.

[0009] Regarding demulsification, it is known that emulsions form in turbulent locations in the production system when oil and water come into contact. Emulsions represent a serious problem for the fuel and petrochemical production chain, such as transportation, storage, and refining. An emulsion is a mixture of two immiscible fluids. One of the two is distributed in the form of droplets identified as the dispersed phase, while the other is known as the continuous phase.

[0010] Oil and water form three types of emulsions: direct, inverse, or multiple. Direct emulsions are characterized by the water being present as a dispersed phase in the crude oil, -water-in-oil- (w / o). In reverse emulsions, water is the continuous phase and the crude oil is dispersed within it, -oil-in-water- (o / w). Multiple or complex emulsions involve small water droplets dispersing within larger oil droplets, with the whole being dispersed in the continuous water phase, -water-in-oil-in-water- (w / o / w), or oil droplets dispersing within larger water droplets, with the whole being dispersed in the continuous oil phase, -oil-in-water-in-oil- (o / w / o).

[0011] The stability of w / o emulsions is mainly due to the presence of amphiphilic molecules in the inferíase of immiscible liquids, such as asphaltenes, resins, or naphthenic acids; these species encapsulate the water droplets and form a film that keeps them dispersed in the oil. O / W emulsions are mainly stabilized by electrostatic repulsion and spherical hindrance; an electrical double layer is present in the inferíase and surface charges, due to the dissociation and adsorption of ions. The repulsion is greater than the van de Waals attraction forces, and this phenomenon keeps the o / w emulsion stable. In the petroleum industry, it is necessary to separate the emulsified phases, and for this purpose, treatment with polymers, surfactants, and other chemicals are recognized as effective and cost-effective techniques.

[0012] W / O demulsifiers are molecules that adsorb at the interface, weaken the intermolecular bonding forces of the species involved in the formation of the film surrounding the droplet, destabilize and break the film, releasing water droplets that can eventually approach and coalesce with each other, forming larger droplets. Large water droplets precipitate due to gravity, while the density difference acts to separate the aqueous from the oily phases. In o / w emulsions, the aim is to neutralize the surface charge of the oil droplets and replace the crude oil species at the interface to promote oil droplet coalescence.

[0013] Surfactants are amphiphilic molecules that have a chemical structure with two clearly differentiated functional groups: a polar region or hydrophilic head and a non-polar region or hydrophobic tail or chain. According to the charge of the head in aqueous solution, the following surfactants are recognized: i) non-ionic (0); i) anionic (-); iii) cationic (+); and iv) zwitterionic (±). A complementary classification of surfactants focuses on the number of polar regions and the number of non-polar regions that the molecule has, as well as their arrangement in space; such that the following main structures are recognized:

[0014] Conventional They present a classic structure composed of a hydrophilic polar head covalently linked to one or two hydrophobic tails.

[0015] Bolaform: They have two polar regions or hydrophilic heads, which can be nonionic, ionic, zwitterionic, or mixtures of these. The heads are linked to one or more hydrophobic chains. Bolaform architecture can therefore be symmetrical or asymmetrical.

[0016] Gemini: Also called dimeric or bipolar, these molecules have two hydrophobic tails of the same type and size and two identical polar heads connected by a spacer (rigid or flexible). Therefore, their architecture necessarily exhibits symmetry.

[0017] Polyvalent: they have multiple hydrophobic chains and several hydrophilic groups interconnected by a multiple spacer.

[0018] The wide variety of surfactants on the market is related to the diversity of possible applications, the multiplicity of possible chemical structures, architectures, and spatial configurations; the origin of their basic components, whether renewable or nonrenewable; and their production, whether through chemical synthesis in the synthesis laboratory or biochemical synthesis in the cells of the species.

[0019] The evolution in surfactant development is moving toward the generation of biodegradable, green chemicals that are harmless to health and environmentally friendly. New and improved amphiphilic molecules do not have to emerge from raw materials derived from petroleum compounds or from conventional chemical synthesis processes; it is easy to deduce then that living beings, prokaryotes and eukaryotes, both their metabolites and their structural components or their biomolecules, constitute a virtually inexhaustible source for the discovery and development of new natural biochemical products.

[0020] Natural products have great virtues and advantages over synthetic chemical products, some of which are: i) very high specificity, granted by the molecular conformation and enantioselectivity of its molecules, i) evolutionary optimization in relation to the biotic and abiotic conditions of the habitat and the selection pressure that the environment has exerted on the species, its molecular components and its metabolites, i¡¡) enormous structural, conformational and electronic diversity of countless biomolecules, iv) source of renewable basics.

[0021] When it comes to searching for and obtaining natural surfactants, microorganisms living in oil fields represent an attractive source for obtaining new, diverse, different and complex biomolecules potentially useful as new surfactants with multifunctionality, biodegradability, low toxicity, stability at high temperature, salinity, and in the presence of divalent cations such as Ca ++ and Mg ++ Of course, they also serve as inspiration for imitating or improving their molecular structure through ad hoc modifications in the chemical synthesis laboratory.

[0022] Indeed, oil fields can support microscopic life, corresponding to extremophile prokaryotes, thermophiles, hyperthermophiles, halophiles, and barophiles, with chemoautotrophic or chemoheterotrophic nutrition and anaerobic respiration. Deeply ancestral species of bacteria and archaea, which inhabit high-temperature, salinity, and pressure environments, such as oil fields, possess a membrane composed of bola-shaped lipid monomers with the following characteristics:

[0023] Bolaform lipids (BL) are composed of three structurally distinct regions: i) a hydrophilic polar region or head, which can be ionic, non-ionic, or zwitterionic; heads in membrane lipids are highly conserved and universal, such as: phosphoethanolamine (PE), phosphocholine (PC), phosphatidylglycerol (PG), and phosphatidic acid (PA);These heads are found in high proportion in the membrane lipids of Bacteria, Archaea and Eucaria, ¡i) a region characterized by the quinal molecule of glycerol, which is formed by the reduction -hydrogenation- of the precursor DHAP in the 2-OH position by NADH and stereospecific enzymatic action of dehydrogenases and acetyltransferases -G1PDH and G3PDH- that form the enantiomers sn-glycerol-1-phosphate (G-1-P) for Archaea and sn-glycerol-3-phosphate (G-3-P) for Bacteria and Eucaria, and i¡¡) a non-polar region characterized by two isoprenoid chains that are linked to glycerol by preferably ether bonds in the sn-2,3 position for Archaea or by alkyl chains or fatty acids that are linked to glycerol by preferably ester bonds in the sn-2,3 position sn-1,2 for Bacteria and Eukarya. In LB, the hydrophobic chains extend along the membrane and form a monolayer and not a conventional bilayer characteristic of Eukarya.

[0024] LB monomers possess particular characteristics that differentiate them from conventional and gemini surfactants. These include the universal presence of the quinone molecule glycerol, in which the heads can have zwitterionic, anionic, nonionic charges, or mixtures thereof, depending on the length and type of hydrocarbon chain. Outside the cell, in aqueous solution, bola-shaped lipid monomers show a strong tendency to self-associate, which leads to the spontaneous formation of supramolecular aggregates with spherical geometry in the form of vesicles. They do not form micelles; self-assembly is mediated by intra- and intermolecular interactions.

[0025] Patent US7798219B1 protects an enhanced recovery process that consists of injecting into the reservoir a modified 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine surfactant where an acyl group is replaced by a phosphoric acid ester, which manages to reduce the interfacial tension to ultra-low values. Patent US20080196892A1 protects an enhanced recovery process that consists of injecting into the reservoir a composition based on enzymes extracted from oil-degrading microorganisms, which reduces the interfacial tension and modifies the wettability of the formation.

[0026] Patent US10415355B1 protects an enhanced recovery process that consists of injecting a nutrient composition into the reservoir to inhibit the growth of an unwanted group of microorganisms and simultaneously stimulate the growth of a specific group of resident microorganisms capable of increasing their interaction with the crude oil rather than with the brine, with the aim that their increased presence in situ significantly decreases the interfacial tension between the oil formation and the water, as well as the wettability of the formation.

[0027] US10889766B2 describes the use of mannosylerythritolipids (MELs) formed by a 4-O-[3-D-mannosyl-pyranosyl-D-erythritol] carbohydrate motif, with acetylation at C4' and C6', linked to fatty acids of different chain lengths, as biodegradable demulsifiers for petroleum emulsions. However, the composition includes a surfactant of non-renewable origin and different from MEL.

[0028] Document WO2013041879 A1 describes the use of the reaction product of alkoxylated poly(vinyl alcohols) and dicarboxylic acids as green, biodegradable, and low-toxicity demulsifiers, taking into account the natural origin of poly(vinyl alcohols) and dicarboxylic acids. However, the composition includes anionic surfactants of non-renewable origin, as well as alkylene oxide block copolymer.

[0029] None of the patents mentioned mention the application of a composition of bolaform lipids as viscoelastic surfactants of natural origin in enhanced hydrocarbon recovery and deemulsification processes. The invention presented surpasses the patents described by virtue of the following main aspects: The LB composition comes from biochemical synthesis, is a biodegradable, non-toxic, environmentally friendly product; LB is not a microorganism that requires nutrients, does not modify the chemical composition of the oil, and does not form micelles or emulsions. In aqueous solution, it forms nanometric-sized vesicles called liposomes or archaeosomes; the LB composition has viscoelastic, dispersing, deemulsifying, wettability-modifying, viscosity-reducing, and interfacial tension properties.It shows stability, compatibility and functionality under conditions of high temperature, salinity, divalent ions, pH variations; functional in carbonate reservoirs, sandstones or mollites, shales or heterogeneous lithographies.

[0030] BRIEF DESCRIPTION OF THE FIGURES

[0031] Figure 1. Thermogravimetric analysis with a ramp of 10 °C / min, and a temperature range of 25 to 510 °C showing the maximum stability at 220 °C of the bolaform lipid composition.

[0032] Figure 2. Micrograph of Thermotoga hypogea taken with transmission electron microscopy.

[0033] Figure 3. Image showing the change in contact angle on impregnated Bedford sheet immersed in brine a) brine without LB and b) brine with 0.005 wt % of the LB composition.

[0034] Figure 4. Image showing the appearance of the Amott cells after 24 h from the start of the comparative study of spontaneous imbibition with impregnation in Bedford limestone to irreducible water, where the LB composition shows the highest oil recovery at the lowest concentration.

[0035] Figure 5. Graph showing oil production behavior at reservoir conditions, where bolaform lipid composition increases oil recovery by up to 23%, after recovering 61% with surfactant-free brine. Figure 6. Image showing an oil-flooded Bedford calcite lamella with LB composition and its desorption by a macroscopic drag, stretching, and droplet formation effect driven by the presence of viscoelastic bolaform lipids.

[0036] Figure 7. Shows the spectral data and structure of the lipid bolaform (2R,36R)- 2-methylene-phosphatidic acid-36-methylene-phosphatylglycerol- 19,20,53,54-tetramethyl- 1,4,35,38- tetraoxocyclooctahexacosane-34,68-dione.

[0037] DETAILED DESCRIPTION OF THE INVENTION

[0038] The present invention provides a novel composition of bolaform lipids as natural surfactants that show high performance at very low concentrations in petroleum industry processes; either as demulsifying agents or as surfactants for enhanced hydrocarbon recovery; the LB composition has viscoelastic, dispersing, demulsifying, wettability-modifying, viscosity-reducing, and interfacial tension properties; stability, compatibility, and functionality under conditions of high temperature, high total dissolved solids, divalent ion, and salinity content. Functionality in carbonate reservoirs, sandstones, mudstones, shales, or in heterogeneous lithologies.

[0039] Bolaform lipids form the cell membrane of deeply ancestral bacterial species, mostly anaerobic and (hyper)thermohalophilic. LB monomers are characterized by being composed of three structurally distinct regions; i) two zwitterionic heads, anionic or non-ionic, where both heads can be the same or different; i) a region characterized by the presence of the quinone molecule glycerol, iii) a non-polar region characterized by two alkyl chains that are linked to the glycerol by ether or ester bonds in the sn-1,2 position. The two alkyl chains extend along the membrane and form a characteristic monolayer membrane and not a conventional bilayer membrane characteristic of eukaryotes.

[0040] Currently, LBs are receiving increased attention as they have demonstrated significant advantages, such as stability, compatibility, and functionality under conditions of high temperature, salinity, dissolved solids, the presence of divalent ions, pH variations, and high resistance to oxidation. The LBs described in the present invention could be used similarly to other surfactants, for example, as detergents, dispersants, wetting agents, or foaming agents; the most common use is as versatile drug nanocarriers. To advance the application of LBs, the following considerations must be addressed:

[0041] • They occur in a complex environment, complex culture media and uncontrolled shake flask cultures and are therefore very heterogeneous,

[0042] • LB composition cannot be controlled,

[0043] • Large quantities are required, in volumes of tons,

[0044] • They cannot be produced profitably.

[0045] Accordingly, the present invention provides a homogeneous and reproducible composition of tailor-made LBs for the petroleum industry considering the following important aspects: i) choice, production and maintenance of the master cell line, and targeted stimulation of the biochemical pathways responsible for LB synthesis; i) increase in LB production, through growth of the selected population using ad-hoc, low-cost, metabolically directed, controlled and kinetically characterized culture medium, iii) recovery, separation, isolation and purification of LBs, iv) formulation and conditioning of LBs for their final application.Bolaform lipids possess specific characteristics, highly differentiated chemically, structurally, and conformationally from Gemini surfactants and other conventional surfactants. They do not form micelle aggregates and therefore do not form emulsions of any kind in any solvent. They are physically and chemically stable; compatible and multifunctional under variable reservoir conditions; and offer high performance at very low concentrations.

[0046] Thermal stability', bolaform lipids have a macrocyclic structure as a thermoadaptive strategy to environments where the temperature is above 65 ° C, the structure and molecular architecture of LB provides microorganisms with a strong packing of LB monomers that form the membrane and reduce its fluidity. The ether bonds and the length of the hydrophobic chain also contribute to the stability of LB in high temperature environments. In this invention, thermogravimetry tests demonstrated that the LB composition is stable at a maximum temperature of 220 ° C. Figure 1 shows the thermogravimetric analysis graph with a ramp of 10 ° C / min, and a temperature range of 25 to 510 ° C and showing stability up to 220 ° C of the bolaform lipid composition.

[0047] Salinity stability: LB monomers possess a structure, molecular conformation, and macrocyclic architecture that present a very low ion permeability coefficient, making the LB composition stable in highly saline environments and in the presence of divalent cations. In vitro tests demonstrated that the LB composition is stable at a maximum salinity of 300,000 ppm at 25°C.

[0048] pH Stability: The composition of LB is stable and functional across the entire pH spectrum, this is due to the ether or ester bonds of the hydrocarbon chains to the glycerol molecule, as well as the presence of zwitterionic heads with neutral charge, which can be in equilibrium in the presence of [H + ] as well as with [OH'] ions.

[0049] Storage stability: LBs have saturated alkyl chains and are therefore not oxidized by air. Consequently, they can be stored at room temperature, without deterioration, for years. More specifically, the present invention provides a method for producing a bolaform lipid composition from a cell culture of a thermohalophilic anaerobic microorganism comprising: a) Growing the bolaform lipid-producing microorganism in an optimized culture medium in a controlled environment provided by a bioreactor. The present invention can be applied in continuous or batch mode, preferably in batch mode. The preferred temperature for the growth of the species is in the range of 70°C to 80°C.b) Standardize the anaerobic inoculum based on the recovery of cells in the pre-exponential growth time, after at least four subcultures, and preferably comprising 10% to 20% of the total fermentation volume, a preferred concentration of 10. 4cells / mL, and a preferred concentration of 0.3 g / L of biomass determined as protein. c) Grow the inoculum from step (b) in a bioreactor, with a culture medium comprising carbohydrates as a carbon source, preferably xylose, glucose or molasses at a concentration between 10 and 25 g / L; a reducing agent as an electron acceptor, preferably sodium sulfide or sodium thiosulfate at a concentration between 2 and 5 g / L; a pH buffer, preferably sodium bicarbonate at a concentration between 3 and 5 g / L. In addition to casein peptone, yeast extract and cysteine ​​as growth cofactors and nitrogen source. As well as salts and divalent ions. d) Supply the bioreactor with a continuous flow of N2, to maintain the anaerobic condition, and agitation, preferably in turbulent regime, to favor the exchange of matter and energy.e) Recovering the bolaform lipids by separating the cells from the culture medium in the late stationary phase of growth; resuspending the cells in a buffer support medium; subsequently lysing them, preferably by ultrasound. f) Extraction and isolation of LB, preferably with a mixture of selected solvents, such as: water, hexane, chloroform, acetone, diethyl ether, methanol, ethanol, ethyl acetate, dichloromethane, and mixtures of these; more specifically and preferably chloroform-methanol-water. g) Storing the bolaform lipid composition in chloroform-methanol and darkness. h) Separation of organic phases from aqueous, recovery of the organic phase containing the LB monomers in the desired proportion, subsequent evaporation of the organic phase to dryness to obtain solvent-free LB, and carrying out the ad hoc solvent formulation for the desired application.

[0050] The lipid-producing thermohalophilic anaerobic microorganism bolaform is selected from the domain Bacteria, of the phylum Termotogae of the order Thermotogales, of the genera that include Geotoga, Kosmotoga, Fervidobacterium, Mesotoga, Petrotoga and Thermotoga, more specifically and preferably Thermotoga hypogea. A transmission electron micrograph of the species Thermotoga hypogea is presented in Figure 2.

[0051] The selected organism can be obtained, for example, from the Japanese Collection of Microorganism Strain, Biological Resource Center, National Institute of Technology and Evaluation, NBRC, catalogue number NBRC 106472. An alternative source of supply is, for example, the Deutsche Sammlung von Mikroorganismen und Zellkulturen, DSMZ, catalogue number DSM 11164. (Hyper)thermophilic members of the phylum Thermotogae of the order Thermotogales possess bola-shaped lipids that are characterized by a non-polar region consisting of two saturated alkyl chains ranging from C2 to C34, with methyl substituents on half of the chains; the chains can form cyclic (I) and (II) or open (III) structures; have both the same (I) and (III) or different carbon numbers (II); The alkyl chains are attached to glycerol at the sn-1,2 position by ester or ether bonds.At both ends (I), (II) and (III) is the polar region or head of the LB monomers, the heads can be both zwitterionic (IV) and (V), anionic (VI) and (VII) or non-ionic (VIII), or have different combinations of the polar regions or heads mentioned.

[0052] The molecular structure of the bolaform lipids object of the present invention is presented below. Where the alkyl chains can be linked to the quinal glycerol molecule in the following ways: tetraether triether / monoester triether / monoester dietether / diester triester / monoether tetraester Where X1 and X2 in structures (I), (II) and (III) is the polar head, both heads can be the same or different.

[0053] Xi: (IV); (V); (VI); (Vile); (VIII)

[0054] X2: (IV); (V); (VI); (Vile); (VIII)

[0055] (IV) (V) phosphoethanolamine (PE) phosphocholine (PC) phosphatidic acid (PA) phosphatidylglycerol (PG)

[0056] (VIII) oc,D-(1 -^4)-glucopyranosyl EXAMPLES

[0057] The following description and examples illustrate certain preferred embodiments and aspects of the present invention, and should not be construed as limiting the scope thereof, since many variations are possible.

[0058] EXAMPLE 1

[0059] This section describes the biotechnology for producing bolaform lipids from Thermotoga hypogea, with a specific and reproducible composition based on strict control of process parameters. Examples include temperature, inoculum quality and quantity, agitation conditions and strategies for maintaining anaerobic conditions during fermentation, culture medium and preparation method, and / or growth phase and rate for cell harvesting. An example of the biotechnological production route and the critical parameters for achieving the desired composition with strict quality control for reproducible LB composition are described below.

[0060] Prior to cultivation in the bioreactor, precultures of the ilized Thermotoga hypogea NBRC 106472 strain were carried out in serological bottles. They were incubated at 70°C, with shaking at 150 rpm, and culture medium 1150 NITE-NBRC was used. Once the strain was reactivated, modifications were made to the culture medium until the best support medium and culture conditions were found that favored LB production, growth rate, amount of biomass, and physiological activity of the strain; as well as the economy of the process. Table 1 shows an example of the composition of the culture medium for obtaining customized bolaform lipids from Thermotoga hypogea for application in the petroleum industry.

[0061] The preculture is aseptically transferred to a bioreactor, which contains the culture medium described in Table 1. The volume of inoculum used for fermentation corresponds, preferably, to 10% of the working volume and the initial cell concentration is preferably X10. 4 , and 0.3 g / L of biomass determined as protein. Fermentation is preferably carried out using batch cultures; N2 is bubbled through the sparger at a preferred flow rate of 50 mL / min; the medium is stirred at turbulent flow; the preferred operating temperature is maintained at 70 e C

[0062] 5 and is operated without pH control or adjustment.

[0063] TABLE 1. COMPOSITION OF THE CULTURE MEDIUM

[0064] Component Concentration

[0065] K2HPO4 0.28 g / L n NH4CI 0.5 g / L U NaCl 1 1.8 g / L

[0066] KCI 0.812 g / L

[0067] BaCI 0.044 g / L

[0068] MgCI22.07 g / L

[0069] CaCI20.44 g / L

[0070] FeCI20.05 mg / L

[0071] MnCI20.9 mg / L

[0072] CuCI20.01 mg / L 5 AICI3 0.21 mg / L

[0073] ZnCI20.01 mg / L

[0074] Peptone-casein 4 g / L

[0075] Yeast extract 4 g / L

[0076] Cysteine ​​- HCI 1 g / L

[0077] Molasses 22.8 g / L

[0078] NaHCO33.64 g / L

[0079] Na2S2O3 4.60 g / L

[0080] For the production of tailor-made LBs for the petroleum industry, cells are harvested in late stationary phase, after at least 3 generation times; they are concentrated in a buffer solution, and lysed by ultrasound. 5 The bolaform lipids are preferably extracted with a chloroform-methanol-water mixture. The organic phase is separated and evaporated with nitrogen gas to obtain the solvent-free lipid extract, resuspended in a chloroform-methanol mixture, and stored in the dark until imminent use. EXAMPLE 2

[0081] Contact angle, the effect of the lipid ball-shape composition on wettability is performed indirectly considering the contact angle measurement; a Krüss DSA100 drop shape analyzer equipment is used, crude oil from a Mexican oil well in the northern region is used, and synthetic brine whose characteristics are presented in tables 2 and 3. 1x1x0.5 cm Bedford calcite sheets are conditioned in an aging cell, the calcite sheets are saturated with crude oil at 2500 psi and 80 ° C for eight days. An oil-saturated sheet is flooded with synthetic brine, the LB composition is added at a concentration of 0.005% by weight at room temperature. This is done in triplicate.

[0082] The contact angle of the oil with respect to the rock changes after 2 hours from the start of the test. The oil desorbs from the surface in the form of small drops with an angle less than 180°, more precisely 75°, indicating a change in the wettability of the surface, as observed in Figure 3, where a change in the contact angle is observed on an impregnated Bedford sheet immersed in brine. Small drops continuously detach from the surface and spontaneously move towards the oil bank and become integrated into it. Desorption in the form of droplets, rather than threads, indicates viscoelastic behavior driven by the presence of LB, as well as a substantial decrease in interfacial tension.

[0083] TABLE 2. CHARACTERISTICS OF CRUDE OIL FROM A MEXICAN FIELD IN THE NORTHERN REGION asphaltenes saturates aromatics resins acid number total gravity

[0084] (% by weight) (% by weight) (% by weight) (mgKOH / g) °API

[0085] 23.43 15.63 28.73 32.21 0.13 15.18 density = 958.685 kg / m 3 ; viscosity = 2401.59 cP @ 25°C. TABLE 3. SYNTHETIC BRINE DESIGN BASED ON THE CONGENITAL WATER COMPONENTS OF A MEXICAN RESERVOIR IN THE NORTHERN REGION reagent concentration

[0086] NaCl 29.9953 g / L

[0087] KCI 0.26980 g / L

[0088] CaCh 0.4577 g / L

[0089] MgCI20.2695 g / L

[0090] BaCI20.0665 g / L

[0091] SrCI20.03897 g / L

[0092] MnCI22.0614 mg / L

[0093] FeCI20.1135 mg / L

[0094] CuCI 0.0156 mg / L

[0095] AICI31 .0378 mg / L

[0096] ZnCl20.0208 mg / L

[0097] EXAMPLE S

[0098] Spontaneous imbibition. This evaluation was carried out in Amott cells at 80° for 20 days, four cores of 68 cm length by 3.82 cm diameter and 80 mD permeability of Berea limestone were used, synthetic brine was used, with characteristics presented in table 3, and crude oil with characteristics presented in table 2. The following procedure was performed: i) core cleaning with a mixture of organic solvents, ¡i) core drying, i¡¡) saturation with synthetic brine, i¡¡) brine displacement by injection of crude oil, iv) conditioning in an aging cell at confinement conditions of 2500 psi and 80°C for 11 days, v) removal of excess oil, vi) each of the four cores was introduced into an Amott cell and put in contact with a) synthetic brine without any type of product; b) synthetic brine plus commercial enzyme-based product at a concentration of 3.0% by weight; c) synthetic brine plus zwitterionic gemini surfactant at a concentration of 0.2% by weight; d) synthetic brine plus bolaform lipid natural surfactant composition at a concentration of 0.01% by weight; i) finally, the oil production in each of the four Amott cells was determined. The results are shown in Table 4. TABLE 4. OIL RECOVERY RESULTS BY SPONTANEOUS IMBIBITION IN AMOTT CELLS.

[0099] Product Concentration Porosity Permeability Recovery factor (%) (mD)

[0100] (% by weight) (%)

[0101] Synthetic brine 0 20.05 78.03 7

[0102] Enzymes 3.0 19.53 75.19 17

[0103] Gemini 0.2 19.44 79.99 24

[0104] Ballform lipids 0.01 19.30 79.04 36

[0105] The results of the comparative study of spontaneous imbibition with impregnation in Bedford limestone with irreducible water show that LB produced the highest oil recovery at the lowest concentration, five times greater than that with brine alone and no surfactant. Gemini enzymes and surfactants also increased oil recovery compared to brine alone, but did so to a lesser extent and at a higher product concentration compared to LB. Figure 4 shows the appearance of the Amott cells 24 h after the start of the comparative study. Note the cap, which corresponds to LB completely impregnated with oil desorbed from the rock.

[0106] In this test, as in the contact angle test, oil production occurs in the form of a multiplicity of small drops and not threads. The small oil drops indicate viscoelastic behavior at the interface driven by the presence of LB.

[0107] EXAMPLE 4

[0108] Production at reservoir conditions. Displacement tests are performed at reservoir conditions, including enhanced hydrocarbon recovery assisted by natural lipid-type surfactants. The test exemplified here was carried out at 80°C, injection pressure of 1000 psi, and confining pressure of 2500 psi; and an injector well-producer well system was considered. TABLE 5. RELEVANT PROPERTIES OF CRUDE OIL FROM A MEXICAN FIELD IN THE NORTHERN REGION. asphaltenes saturates aromatics resins acid number total gravity

[0109] (% by weight) (% by weight) (% by weight) (mgKOH / g) °API

[0110] 7.19 41 .98 25.94 24.89 0.15 25.43 density = kg / m 3 ; viscosity = 35 cP @ 25°C.

[0111] Fluids. Crude oil from a Mexican field in the northern region was used; its properties are presented in Table 5; as well as synthetic brine, the composition of which is shown in Table 3.

[0112] Core preparation. A core from a low-permeability Mexican reservoir with a shale / sandstone lithology, measuring 8.89 cm in diameter and 10.03 cm in length, was used. The core was cleaned by successively injecting various pore volumes of hexane, toluene, chloroform, and methylene chloride; then hexane, toluene, and finally a dichloromethane / water mixture, ensuring maximum oil release. The cleaned core was dried in a high-vacuum oven at 100°C. The core characteristics are presented in Table 6.

[0113] TABLE 6. CHARACTERISTICS OF THE CORE FROM A MEXICAN DEPOSIT IN THE NORTHERN REGION.

[0114] Feature _ value

[0115] Pore ​​volume (cm 3 ) 136.30

[0116] Volume of irreducible brine in core (cm 3 ) 23.60

[0117] Oil volume in core (cm 3 ) 112.7

[0118] Porosity (%) 21.89

[0119] Permeability with distilled water (mD) 1 .84

[0120] Permeability with oil + irreducible water (mD) 0.58

[0121] Initial brine saturation (%) 17.32

[0122] Initial saturation of crude oil in core (%) 82.68

[0123] First Stage: Core Conditioning After determining porosity and permeability, the core was saturated with synthetic brine; the saturated core was aged for one week. The rock was then saturated with dead crude oil at a constant rate of 0.5 mL / h until irreducible water saturation was reached; it was aged for three weeks. Secondary Stage: Oil Displacement: Synthetic brine was injected at a constant rate of 0.5 mL / h until oil production stopped and only brine was observed in the collection vessels. This was achieved after 1.5 pore volumes and 61% oil recovery.

[0124] Third stage: enhanced recovery The residual oil saturation value of the secondary recovery stage was determined prior to starting the enhanced recovery. The bolaform lipid composition was injected at a constant flow rate of 0.5 mL / h and a concentration of 0.01% by weight; when oil production reached 80% and after 2.5 pore volumes, the injection of only brine without LB was resumed, thus verifying the benefit of improved oil production due to the LB composition and ruling out any possible damage to the core. Under this 80% condition, production increased to 83% due to the residual presence of LB in the core. Once it was observed and determined that there was no further increase in oil production, the LB composition was injected again, one more pore volume and we observed an increase in production up to 84%, where the test was concluded.The graph in Figure 5 shows the oil production behavior at reservoir conditions, where the composition of bolaform lipids increases oil recovery by up to 23%, after the recovery of 61% with surfactant-free brine.

[0125] Then, under reservoir conditions, a composition of LB is injected, forming nanosized liposomal vesicles; nanoliposomes and LB spontaneously target the oil-brine interface in the bulk and into rock cavities.

[0126] LB monomers have an inherent tendency to self-associate; they form aggregates mediated by intra- and intermolecular interactions, the hydrophobic effect, and hydrogen bonding; the aggregates tend to close in on themselves, grow, and extend. Thus, in aqueous solution, LB monomers aggregate and form vesicles. The vesicles, in turn, aggregate and form cylindrical or tubular structures called worms. The worms, in turn, intertwine and untwist with each other. This leads to a dynamic rearrangement of the supramolecular association of LB. The intertwined worm-like supramolecular structure of LB at the interface provides viscoelastic properties that influence fluid mobility. Flooding with LB retards mobility in fracture and high-permeability zones, where the interface becomes oily and viscosity increases; whereas the viscoelastic interface thins and its viscosity decreases upon entering low-permeability zones.Thus, the preferential fluid flow towards areas of greater permeability is overcome with great effectiveness by the LB composition.

[0127] LBs have a strong electrostatic interaction energy with resins and asphaltenes, so great that they weaken the interactions of these crude oil components with the rock formation and therefore with the cohesive forces of the crude oil to the surface. Once the interaction energy between the oil and the rock is weakened, small oil droplets are released from the formation cavities; a macroscopic drag, stretching, and droplet formation effect occurs, driven by the presence of viscoelastic spherical lipids. The released droplets flow spontaneously toward the oil bank and coalesce there; this macroscopic effect is illustrated in Figure 6.

[0128] LB and asphaltenes form a supramolecular complex with greater energy than the interaction energy of asphaltenes with each other, and therefore prevents their stacking, favors their dispersion, and reduces the viscosity of the oil.

[0129] LB monomers possess chirality and the asymmetric LB wormlike supramolecular structure, which leads to the formation of directional domains and preferential fluidity of discrete crude oil droplets that flow toward and increase the oil bank. The elasticity of the interface maintains the continuity of the oil and aqueous phases, which flow as continuous phases separated from each other. The EOR and demulsification mechanisms of LB are a synergistic effect of reducing interfacial tension, changing wettability, and affecting crude oil mobility. The fluid flow proceeds without clogging or blocking channels, pores, or pore throats; LB improves sweeping, displacement, and recovered oil volume; minimizes formation damage and maintains formation permeability; does not cause preferential paths or fingering, and does not produce emulsions.

[0130] EXAMPLE S

[0131] The objective of this example is to show the effect of LB in demulsifying brine into oil, with different brine / oil ratios.

[0132] Preparation of the emulsion - The emulsion was formed using as fluids an oil from a Mexican field with the characteristics described in Table 2 and brine whose components are listed in Table 3. An IKA labortechnik homogenizer was used and the whole mixture was stirred with an Ultra-Turrax mixer at 11,600 rpm for 2 minutes.

[0133] Demulsification test -, a LB stock solution is prepared in brine, the volume of the LB stock solution is adjusted according to the desired -water / oil- ratio to ensure the same concentration of 0.01% by weight in all treatments. The necessary quantities of brine stock solution plus LB at a concentration of 0.01% by weight and volumes from 80 to 10 ml) and emulsified oil from 20 to 90 ml were introduced into a container, the mixtures were stirred with an Ultra-Turrax type mixer at 11,600 rpm for 2 minutes. Each sample obtained was poured into 100 ml graduated conical-bottom tubes to control and quantify the separation of oil and water at room temperature.

[0134] Water volumes were read after 1 and 24 hours of settling at room temperature. The percentages of water recovered correspond to the volumes of water read in the test tube relative to the volumes of aqueous solutions introduced. Tests were performed in triplicate, and controls without the addition of LB were also prepared. In the controls, brine recovery was less than 90% for all test variables.

[0135] Table 7 presents the brine recovery percentage (%vol) of the brine / oil mixture, calculated based on the total brine content.

[0136] TABLE 7. PERCENT OF BRINE RECOVERED FROM EMULSIFIED CRUDE OIL.

[0137] Tempo i

[0138] .. . Brine / ; oil

[0139] (hours)

[0140] 80 / 20 60 / 40 50 / 50 40 / 60 20 / 80 10 / 90

[0141] 1 >85 >85 >90 >95 >95 >95

[0142] 24 >90 >90 >95 >95 >95 >95

[0143] EXAMPLE 6

[0144] Identification of bolaform lipids The identification of I [bolaform lipids was determined by ultrahigh performance liquid chromatography coupled to electrospray ionization source time-of-flight tandem mass spectroscopy; LC-QTOF / MS / MS-ESI. A WATERS ACQUITY UPLC-I-CLASS-SYNAPT G2 equipment, equipped with a gas collision chamber, was used, grounded for high-energy CID experiments. All analytes were characterized by high-energy LC / QTOF-MS / MS / R / ESI+ CID, mass calibration was performed with a commercial peptide mixture supplied by Waters allowing mass calibration up to m / z 3600. In positive ion mode, mainly [M+H]+ adduct ions of the analyte molecules were detected. The solvent-free bolaform lipid samples were redissolved in a CHChMeOH mixture, 5pL of sample with MeOH / h mobile phase in HCOOH were injected and 90 min runs were performed.Figure 7 shows the spectral data and structure of the lipid bolaform (2R,36R)-2-methylene-phosphatidic acid-36-methylene-phosphatylglycerol-19,20,53,54-tetramethyl-1,4,35,38-tetraoxocyclooctahexacosane-34,68-dione.

Claims

CLAIMS Having described the present invention, it is considered novel and therefore the content of the following clauses is claimed as our property:

1. A composition of I [bolaform lipids characterized in that the I [lipids have a non-polar region formed by two saturated alkyl chains ranging from C28 to C34, with methyl substituents in the middle of the chains; the chains can form cyclic or open structures; both have the same or different number of carbons; the alkyl chains are attached to the glycerol in the sn-1,2 position by ester or ether bonds, giving rise to structures such as: tetraester; triester / monoether; diester / diether; monoester / trieter or tetraether; at both ends of the described structure is the polar region or head, the heads can both be zwitterionic, anionic or non-ionic, or have different combinations of the polar regions or heads stated.

2. The composition according to claim 1, characterized in that the zwitterionic polar head may include primary, secondary or tertiary amines or quaternary ammonium cations, and sulfonate or phosphate groups.

3. The composition according to claim 2, characterized in that the zwitterionic polar head is phosphoethanolamine.

4. The composition according to claim 2, characterized in that the zwitterionic polar head is phosphocholine.

5. The composition according to claim 1, characterized in that the anionic polar head may contain a sulfate or phosphate group.

6. The composition according to claim 5, characterized in that the anionic polar head is phosphatidic acid.

7. The composition according to claim 5, characterized in that the anionic polar head is phosphatidylglycerol.

8. The composition according to claim 1, characterized in that the non-ionic polar head may contain a carbohydrate selected from the group of saccharides.

9. The composition according to claim 8, characterized in that the non-ionic polar head is α,D-(1 -^4)-glucopyranosyl.

10. A method for obtaining bolaform lipids according to claim 1, characterized in that it comprises the following steps: a) Growing a thermohalophilic anaerobic microorganism producing bolaform lipids in a bioreactor in operating mode, preferably discontinuous, and at a preferred temperature of 70 to 80°C. b) Standardizing the anaerobic inoculum based on the recovery of biomass in the pre-exponential growth phase, and comprising 10% to 20% of the total volume of the fermentation, a preferred concentration of 10 4cells / mL, and a concentration of 0.3 g / L of biomass determined as protein. c) Grow the inoculum from step (b) in a culture medium comprising carbohydrates as a carbon source, a reducing agent to maintain the anaerobic condition, and as a final electron acceptor; a pH buffering agent. This at concentrations such that, the increase cell reaches at least three orders of magnitude, the increase in protein is at least five times, with respect to the biomass inoculated in step (b), and until the initial concentration of the carbon source is reduced by at least 80%; d) Additionally, the culture medium of step (c) comprises casein peptone, yeast extract and cysteine as growth cofactors and a nitrogen source in concentrations such that they allow obtaining at least 30% of bolaform lipids in relation to the biomass concentration quantified as protein. e) Supply a continuous flow of N2 during fermentation to maintain the anaerobic condition, and turbulent flow agitation to promote the exchange of matter and energy.f) Recover the bolaform lipids by separating the cells from the late stationary growth phase culture medium, then lysing them using ultrasound; extraction and isolation using a preferred mixture of chloroform-methanol-water.

11. The method according to claim 10, characterized in that the anaerobic, thermohalophilic, lipid-producing, bolaform microorganism is selected from the domain Bacteria, of the Phylum Termotogae of the order Thermotogales of the genera comprising Geotoga, Kosmotoga, Fervidobacterium, Mesotoga, Petrotoga and Thermotoga.

12. The method of claim 10, characterized in that the carbon source is selected from xylose, glucose and / or molasses.

13. The method of claim 10 characterized in that in step (c) the carbon source is at a concentration between 10 and 25 g / L.

14. The method of claim 10 characterized in that the reducing agent and electron acceptor is selected from sodium sulfide or sodium thiosulfate.

15. The method of claim 10, characterized in that in step (c) the reducing agent and electron acceptor is at a concentration of 2 to 5 g / L.

16. The method of claim 10, characterized in that the pH buffer system is selected from sodium bicarbonate or sodium phosphate salts.

17. The method of claim 10, characterized in that in step (c) the pH buffer system is at a preferred concentration of 2 to 4 g / L.

18. A composition according to claim 1, characterized in that it comprises bolaform lipids, plus polar solvents such as: seawater, or treated water or brines with a high content of divalent ions, salts and total dissolved solids or combinations thereof; and / or organic solvents derived from alcohols such as methanol, ethanol, or isopropanol, or mixtures thereof; and / or aromatic solvents such as benzene, toluene, or xylene or combinations thereof.

19. The composition for the enhanced recovery of hydrocarbons according to claim 18, characterized in that the concentration of bolaform lipids is between 0.005 and 0.01% by weight.

20. The use of the composition according to claim 19, for the enhanced recovery of hydrocarbons.

21. The use of the composition according to claim 19, wherein the oil is supplied to an injection well and the oil is recovered in a production well.

22. The use of the composition according to claim 19, wherein it is supplied in the same well that acts as injector and producer.

23. The use of the composition according to claim 19, wherein the salt concentration can reach up to 300,000 ppm.

24. The use of the composition according to claim 19, wherein the concentration of divalent ions can reach up to 170,000 ppm.

25. The use of the composition according to claim 19, wherein the temperature can reach up to 220°C.

26. The composition for demulsification processes and methods of O / W; W / O; W / O / W; O / W / O emulsions, according to claim 18, characterized in that the concentration of bolaform lipids is between 0.005 and 0.01% by weight.

27. The use of the composition according to claim 26, wherein the emulsified crude oil may have a brine content between 10 and 90%, by volume with respect to the volume of the emulsion, the salinity may reach 300,000 ppm, the content of divalent ions and cations may reach up to 180,000 ppm and the temperature may reach 220°C.

28. The use of the composition according to claim 26, wherein the crude oil may be of light, medium, heavy and extra heavy type or mixtures thereof.

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