阅读说明：本技术 复合微生物菌剂、其制备方法及应用 (Compound microbial agent, preparation method and application thereof ) 是由 郑爱萍 夏文杰 杜雪彪 冀楠 石磊 巨世昌 徐绍轩 梁向进 于 2020-05-28 设计创作，主要内容包括：本发明公开了一种复合微生物菌剂、其制备方法及应用。其中,该复合微生物菌剂由微生物和辅料制成,其中,微生物由芽孢杆菌、肠杆菌、乳酸菌和假单胞菌混合制成。本发明的复合微生物菌剂包含特定的四种菌种,能够避免因单功能菌种不耐受环境冲击的缺陷,并且四种菌株之间的功能表达没有受到抑制,且制备方法简单、成本低。根据不同的微生物菌种代谢特性不同,产生的代谢产物不同,形成的驱油采油机理不同,由此也形成了不同的微生物采油工艺,四种微生物相互协同,产生了很好的技术效果。(The invention discloses a compound microbial agent, a preparation method and application thereof. The compound microbial agent is prepared from microorganisms and auxiliary materials, wherein the microorganisms are prepared by mixing bacillus, enterobacter, lactobacillus and pseudomonas. The compound microbial agent comprises four specific strains, can avoid the defect that the single-function strains cannot tolerate environmental impact, has no inhibition on functional expression among the four strains, and has simple preparation method and low cost. According to different microbial species, the metabolic characteristics of the produced metabolites are different, and the formed oil displacement and oil extraction mechanisms are different, so that different microbial oil extraction processes are formed, and the four microbes are synergistic to produce a good technical effect.)

1. The compound microbial agent is characterized by being prepared from microorganisms and auxiliary materials, wherein the microorganisms are prepared by mixing bacillus, enterobacter, lactobacillus and pseudomonas.

2. The complex microbial inoculant according to claim 1, wherein the ratio of the number of bacillus, enterobacter, lactic acid bacteria and pseudomonas is (1-6): (1-6): 1-6).

3. The compound microbial inoculant according to claim 1, wherein the auxiliary material is a microbial liquid culture medium.

4. The complex microbial inoculant according to claim 3, wherein the liquid culture medium is prepared from the following raw materials in percentage by weight: 0.5-2.5% corn syrup, 0.5-2.5% vitamins, 0.5E5% of vegetable oil and 0.1-0.5% of KH₂PO₄0.01 to 0.1% of NaNO₃0.1-0.2% of yeast powder, 0.01-0.1% of benzene, 0.1-0.2% of xanthan gum and the balance of water.

5. The complex microbial inoculant according to claim 3 or 4, wherein the pH of the liquid culture medium is between 7 and 8.

6. A method for preparing the complex microbial inoculant according to any one of claims 1 to 5, comprising the following steps: respectively inoculating bacillus, enterobacter, lactobacillus and pseudomonas into a liquid culture medium for culture, and then taking the culture medium containing the strains as the bacillus according to the strain quantity proportion: enterobacteria: mixing lactobacillus and pseudomonas in the ratio of (1-6) to obtain the compound microbial agent.

7. The preparation method according to claim 6, wherein the liquid culture medium is prepared from the following raw materials in percentage by weight: 0.5-2.5% of corn syrup, 0.5-2.5% of vitamins, 0.5-5% of vegetable oil, and 0.1-0.5% of KH₂PO₄0.01 to 0.1% of NaNO₃0.1-0.2% of yeast powder, 0.01-0.1% of benzene, 0.1-0.2% of xanthan gum and the balance of water.

8. The production method according to claim 7, wherein the pH of the liquid medium is 7 to 8.

9. The method according to claim 6, wherein the culturing of the Bacillus, Enterobacter, lactic acid bacteria, and Pseudomonas is carried out at 30 to 40 ℃ for 36 to 50 hours.

10. The method according to claim 6, wherein the amounts of the Bacillus, Enterobacter, lactic acid bacteria, and Pseudomonas are 0.1 to 10%, 0.2 to 10%, and 0.1 to 10% by weight, respectively, of the liquid medium;

preferably, the inoculation amounts of the bacillus, the enterobacteria, the lactic acid bacteria and the pseudomonas are respectively 1%, 3% and 4% of the weight fraction of the liquid culture medium.

11. Use of the complex microbial inoculant of any one of claims 1 to 5 in oil recovery;

preferably, the complex microbial inoculant is injected into the oil well in one or more steps.

Technical Field

The invention relates to the technical field of microbial oil recovery, in particular to a compound microbial agent, and a preparation method and application thereof.

Background

Along with the rapid development of economy in China, the demand for petroleum is increasing day by day, the shortage of petroleum is also getting worse, the proportion of unconventional oil gas is getting larger and larger, and the thickened oil is one of five unconventional resources at present.

Most of petroleum resources left in the world are heavy oil resources, the resource amount is more than 8000 hundred million tons, but heavy oil exploitation mainly takes steam injection thermal exploitation as main development, and the problems of extremely high development cost, high energy consumption and short effective period exist, particularly in the middle and later stages of development, the oil-steam ratio is greatly reduced, the cost is greatly increased, large-scale low-efficiency heavy oil cannot be economically and effectively developed, and the problem of how to improve the quality and the efficiency is urgently to be solved.

The Microbial Enhanced Oil Recovery (MEOR) technology is a biological technology for improving the yield of crude oil by using the life activities and metabolites of microbes in oil reservoirs, has the characteristics of wide application range, low implementation cost, remarkable economic benefit and the like, and is increasingly paid more attention by people.

The mechanism required by the microbial oil displacement for the development of the thick oil mainly comprises the following mechanisms: gas generated in the microbial growth and metabolism process increases formation pressure and reduces the viscosity of crude oil; the biosurfactant and the organic solvent are generated to reduce the oil-water interfacial tension and improve the oil washing efficiency; degrading heavy components of the crude oil to reduce the viscosity of the crude oil, improve the fluidity of the crude oil, and the like.

The metabolic characteristics of different microbial strains are different, the produced metabolites are different, and the oil displacement mechanisms of the microbial strains are also different, so that different microbial oil recovery processes are formed.

Disclosure of Invention

The invention aims to provide a compound microbial agent, a preparation method and application thereof, and aims to solve the technical problem that the complex preparation or single strain preparation of the existing compound microbial agent in the prior art is not resistant to environmental impact.

In order to achieve the above object, according to one aspect of the present invention, there is provided a complex microbial inoculant. The compound microbial agent is prepared from microorganisms and auxiliary materials, wherein the microorganisms are prepared by mixing bacillus, enterobacter, lactobacillus and pseudomonas.

Further, the number ratio of bacillus, enterobacter, lactic acid bacteria and pseudomonas is (1-6): (1-6).

Furthermore, the auxiliary material is a microorganism liquid culture medium.

Further, the liquid culture medium is prepared from the following raw materials in percentage by weight: 0.5-2.5% of corn syrup, 0.5-2.5% of vitamins, 0.5-5% of vegetable oil and 0.1-0.5% of KH₂PO₄0.01 to 0.1% of NaNO₃0.1-0.2% of yeast powder, 0.01-0.1% of benzene, 0.1-0.2% of xanthan gum and the balance of water.

Further, the pH of the liquid medium is 7 to 8.

According to another aspect of the present invention, there is provided a method for preparing any one of the above-mentioned complex microbial agents. The preparation method comprises the following steps: respectively inoculating bacillus, enterobacter, lactobacillus and pseudomonas into a liquid culture medium for culture, and then taking the culture medium containing the strains as the bacillus according to the strain quantity proportion: enterobacteria: mixing lactobacillus and pseudomonas in the ratio of (1-6) to obtain the composite microbial agent.

Further, the liquid culture medium is prepared from the following raw materials in percentage by weight: 0.5-2.5% of corn syrup, 0.5-2.5% of vitamins, 0.5-5% of vegetable oil and 0.1-0.5% of KH₂PO₄0.01 to 0.1% of NaNO₃0.1-0.2% of yeast powder, 0.01-0.1% of benzene, 0.1-0.2% of xanthan gum and the balance of water.

Further, the pH of the liquid medium is 7 to 8.

Further, the conditions for culturing the bacillus, the enterobacteria, the lactic acid bacteria and the pseudomonas are all culturing for 36-50 h at 30-40 ℃.

Further, the inoculation amounts of the bacillus, the enterobacter, the lactobacillus and the pseudomonas are respectively 0.1-10%, 0.2-10% and 0.1-10% of the weight fraction of the liquid culture medium; preferably, the inoculation amounts of bacillus, enterobacter, lactobacillus and pseudomonas are respectively 1%, 3% and 4% of the weight fraction of the liquid culture medium.

According to a further aspect of the present invention, there is provided a use of any one of the above-mentioned complex microbial agents in oil recovery; preferably, the complex microbial inoculant is injected into the oil well in one or more portions.

The compound microbial agent comprises four specific strains, can avoid the defect that the single-function strains cannot tolerate environmental impact, has no inhibition on functional expression among the four strains, and has simple preparation method and low cost. According to different microbial species, the metabolic characteristics of the produced metabolites are different, and the formed oil displacement and oil extraction mechanisms are different, so that different microbial oil extraction processes are formed, and the four microbes are synergistic to produce a good technical effect.

Drawings

The accompanying drawings, which are incorporated in and constitute a part of this application, illustrate embodiments of the invention and, together with the description, serve to explain the invention and not to limit the invention. In the drawings:

fig. 1 shows a visual chart (upper chart) of crude oil emulsification and dispersion experiments performed by four bacteria alone and mixed bacteria in example 6 and a particle size analysis chart of crude oil emulsification by the four bacteria alone and mixed bacteria.

Detailed Description

It should be noted that the embodiments and features of the embodiments in the present application may be combined with each other without conflict. The present invention will be described in detail below with reference to the embodiments with reference to the attached drawings.

According to an exemplary embodiment of the present invention, a complex microbial inoculant is provided. The compound microbial agent is prepared from microorganisms and auxiliary materials, wherein the microorganisms are prepared by mixing Bacillus (Bacillus), enterobacter (Enterobacter), lactobacillus (Lactobacillus) and Pseudomonas (Pseudomonas).

The compound microbial agent comprises four specific strains, can avoid the defect that the single-function strains cannot tolerate environmental impact, has no inhibition on functional expression among the four strains, and has simple preparation method and low cost. According to different microbial species, the metabolic characteristics of the produced metabolites are different, and the formed oil displacement and oil extraction mechanisms are different, so that different microbial oil extraction processes are formed, and the four microbes are synergistic to produce a good technical effect.

Preferably, the number ratio of bacillus, enterobacter, lactic acid bacteria and pseudomonas is (1-6): (1-6). In this case, the synergistic effect among the four microorganisms can be better embodied.

According to a typical embodiment of the invention, the auxiliary material is a microbial liquid culture medium, namely, the cultured microbial liquid can be directly mixed to serve as a compound microbial agent, and the preparation method is simple and low in cost.

Preferably, the liquid culture medium is prepared from the following raw materials in percentage by weight: 0.5-2.5% of corn syrup, 0.5-2.5% of vitamins, 0.5-5% of vegetable oil and 0.1-0.5% of KH₂PO₄0.01 to 0.1% of NaNO₃0.1-0.2% of yeast powder, 0.01-0.1% of benzene, 0.1-0.2% of xanthan gum and the balance of water. The four kinds of bacteria can be well propagated in the liquid culture medium of the above components, and the raw materials areWide source, easy obtaining, low preparation cost and easy industrialized production. More preferably, the pH of the liquid medium is 7 to 8, and the strain has a strong ability to proliferate at this pH range.

According to a typical embodiment of the present invention, there is provided a method for preparing the complex microbial agent. The preparation method comprises the following steps: respectively inoculating bacillus, enterobacter, lactobacillus and pseudomonas into a liquid culture medium for culture, and then taking the culture medium containing the strains as the bacillus according to the strain quantity proportion: enterobacteria: mixing lactobacillus and pseudomonas in the ratio of (1-6) to obtain the composite microbial agent.

The compound microbial agent comprises four specific strains, can avoid the defect that the single-function strains cannot tolerate environmental impact, has no inhibition on functional expression among the four strains, and has simple preparation method and low cost. According to different microbial species, the metabolic characteristics of the produced metabolites are different, and the formed oil displacement and oil extraction mechanisms are different, so that different microbial oil extraction processes are formed, and the four microbes are synergistic to produce a good technical effect.

Preferably, the liquid culture medium is prepared from the following raw materials in percentage by weight: 0.5-2.5% of corn syrup, 0.5-2.5% of vitamins, 0.5-5% of vegetable oil and 0.1-0.5% of KH₂PO₄0.01 to 0.1% of NaNO₃0.1-0.2% of yeast powder, 0.01-0.1% of benzene, 0.1-0.2% of xanthan gum and the balance of water. In the liquid culture medium with the components, the four bacteria can be well propagated, and the raw materials are wide in source and easy to obtain, low in preparation cost and easy for industrial production. More preferably, the pH of the liquid medium is 7 to 8, and the strain has a strong ability to proliferate at this pH range.

Preferably, the conditions for culturing the bacillus, the enterobacter, the lactobacillus and the pseudomonas are all culturing for 36-50 h at 30-40 ℃, the culturing temperature condition is not harsh, and the large-scale culture is easy. More preferably, the inoculation amounts of the bacillus, the enterobacter, the lactobacillus and the pseudomonas are respectively 0.1-10%, 0.2-10% and 0.1-10% of the weight fraction of the liquid culture medium, so that the activation and the value increase of the strains are facilitated, and further preferably, the inoculation amounts of the bacillus, the enterobacter, the lactobacillus and the pseudomonas are respectively 1%, 3% and 4% of the weight fraction of the liquid culture medium.

According to a typical embodiment of the invention, the application of the compound microbial agent in oil extraction is provided; preferably, the complex microbial inoculant is injected into the oil well in one or more portions.

The following examples are provided to further illustrate the advantageous effects of the present invention.

Example 1

A preparation method of a compound microbial agent comprises the following steps:

(1) preparing a liquid culture medium, wherein the culture medium is prepared from the following raw materials in percentage by weight: 0.5% corn syrup, 0.5% vitamins, 2% vegetable oil, 0.1% KH₂PO₄0.01% of NaNO₃0.1% of yeast powder, 0.01% of benzene, 0.1% of xanthan gum and the balance of water; adjusting the pH value of the culture medium to 7, and sterilizing; wherein, benzene can be used as a carbon source and a solubilizer;

(2) respectively carrying out inoculation culture according to the ratio that the inoculation amounts of bacillus, enterobacter, lactobacillus and pseudomonas are respectively 1.456 percent of the weight fraction of the liquid culture medium, wherein the culture time is 36 hours, and the culture temperature is 30 ℃;

(3) and mixing the cultured strains according to the ratio of 1:1:1:1 to obtain the compound microbial agent.

Example 2

A preparation method of a compound microbial agent comprises the following steps:

(1) preparing a liquid culture medium, wherein the culture medium is prepared from the following raw materials in percentage by weight: 2.5% corn syrup, 2.5% vitamins, 5% vegetable oil, 0.5% KH₂PO₄0.1% of NaNO₃0.2% of yeast powder, 0.1% of benzene, 0.2% of xanthan gum and the balance of water; adjusting the pH value of the culture medium to 7, and sterilizing; wherein benzene can be used as both carbon source and carbon sourceIs a solubilizer;

(2) respectively carrying out inoculation culture according to the ratio that the inoculation amounts of bacillus, enterobacter, lactobacillus and pseudomonas are respectively 10 percent of the weight fraction of the liquid culture medium, wherein the culture time is 36h, and the culture temperature is 30 ℃;

(3) and mixing the cultured strains according to the ratio of 1:1:1:1 to obtain the compound microbial agent.

Example 3

A preparation method of a compound microbial agent comprises the following steps:

(1) preparing a liquid culture medium, wherein the culture medium is prepared from the following raw materials in percentage by weight: 1.5% corn syrup, 1.5% vitamins, 2% soybean oil, 0.2% KH₂PO₄0.05% of NaNO₃0.15% of yeast powder, 0.05% of benzene, 0.15% of xanthan gum and the balance of water; adjusting the pH value of the culture medium to 7.5, and sterilizing; wherein, benzene can be used as a carbon source and a solubilizer;

(2) respectively carrying out inoculation culture according to the inoculation amounts of the bacillus, the enterobacter, the lactobacillus and the pseudomonas which are respectively 2 percent of the weight fraction of the liquid culture medium, wherein the culture time is 45h, and the culture temperature is 35 ℃;

(3) and mixing the cultured strains according to the ratio of 2:1:1:4 to obtain the compound microbial agent.

Example 4

A preparation method of a compound microbial agent comprises the following steps:

(1) preparing a liquid culture medium, wherein the culture medium is prepared from the following raw materials in percentage by weight: 0.5% corn syrup, 0.5% vitamins, 0.5% vegetable oil, 0.1% KH₂PO₄0.01% of NaNO₃0.1% of yeast powder, 0.01% of benzene, 0.1% of xanthan gum and the balance of water; adjusting the pH value of the culture medium to 7.5, and sterilizing; wherein, benzene can be used as a carbon source and a solubilizer;

(2) respectively carrying out inoculation culture according to the ratio that the inoculation amounts of bacillus, enterobacter, lactobacillus and pseudomonas are respectively 5 percent of the weight fraction of the liquid culture medium, wherein the culture time is 45h, and the culture temperature is 35 ℃;

(3) and mixing the cultured strains according to a ratio of 4:1:1:2 to obtain the compound microbial agent.

Example 5

A preparation method of a compound microbial agent comprises the following steps:

(1) preparing a liquid culture medium, wherein the culture medium is prepared from the following raw materials in percentage by weight: 1.0% corn syrup, 2.13% vitamins, 0.45% KH₂PO₄4 percent of rapeseed oil and 0.1 percent of NaNO₃0.4% of yeast powder, 0.11% of benzene, 0.12% of xanthan gum and the balance of water; adjusting the pH value of the culture medium to 8, and sterilizing; wherein, benzene can be used as a carbon source and a solubilizer;

(2) respectively carrying out inoculation culture according to the ratio that the inoculation amounts of bacillus, enterobacter, lactobacillus and pseudomonas are respectively 5 percent of the weight fraction of the liquid culture medium, wherein the culture time is 50h, and the culture temperature is 40 ℃;

(3) and mixing the cultured strains according to the ratio of 1:1:3:6 to obtain the compound microbial agent.

The experimental results are as follows: injecting the compound microbial agent into a low-yield heavy oil well at one time or in batches, determining the injection amount according to the oil-water property and the geological oil reservoir characteristics of a test block, wherein the injection amount is suitable for heavy oil with underground viscosity of below 5000 centipoise, the concentration of the injected bacteria is 1-10%, and if the injection is performed in batches, the injection times are 4-12 times a year; selecting low-yield heavy oil wells in Hongkong and Xinjiang oil fields, performing microorganism single-well huff and puff test, adding 200 parts of compound microorganism bacterium agent (with the concentration of 10) by adopting an oil pipe adding method⁸cfu/ml), the well is opened to recover the production after 20 days of closing the well, the daily oil production is increased from 0.2-1.5 t/day to 1.6-2.69 t/day, and the oil increasing effect is obvious.

Table 1 shows the experimental results of culture media containing Bacillus, Enterobacter, lactic acid bacteria and Pseudomonas in different proportions:

TABLE 1 Experimental Effect of complex bacteria in different proportions

TABLE 2 Experimental comparison of Single Strain and Complex microbial Agents

The field application method comprises the following steps:

the composite microbial agent is injected into a heavy oil well at one time or in batches, the injection amount is determined according to the oil-water property and the geological oil reservoir characteristics of a test block, the composite microbial agent is suitable for heavy oil with the underground viscosity of below 20000 centipoise, the concentration of the injected bacteria is 1-10%, and if the composite microbial agent is injected in batches, the injection times are 4-12 times a year.

Example 6

The synergistic effect of the four strains improves the oil extraction function of the composite microbial inoculum.

Crude oil emulsification and dispersion experiments (upper diagram in figure 1) are carried out on four independent bacteria and mixed bacteria, and particle size analysis (lower diagram in figure 1) of the corresponding four independent bacteria and mixed bacteria emulsified crude oil is carried out, so that the effect difference of the single bacteria is large, a good coordination effect is formed after the four microbial agents are mixed, the emulsified particle size is uniform, the dispersion is fine, and the mixed bacterial strain has the best emulsification effect on the crude oil; from the viewpoint of the emulsification grade (tables 3 and 4), the emulsification grade of the mixed bacteria is superior to that of the single bacteria in different culture media.

Wherein, the emulsification experiment step: 30ml of the bacterial liquid is inoculated into a shake flask, and 300ml of the culture medium and 60 g of crude oil are filled in a 1000ml shake flask and are cultured for 7 days in a 36 ℃ shaking table.

TABLE 3

	Pseudomonas sp	Bacillus	Enterobacter	Lactic acid bacteria	Mixed bacteria
						Emulsification rating in Medium (i)	++++	+++	+++	++	+++++
Emulsification rating in Medium-	++++	++	+++	++	+++++

Note: the culture medium I and the culture medium II are conventional inorganic salt culture media using crude oil carbon sources, and the difference between the culture medium I and the culture medium II is that 0.1 percent of glucose is added in the culture medium II; preparing the mixed bacteria in a ratio of 1:1:1: 1; crude oil emulsification rating was determined by visual observation according to industry conventions.

TABLE 4

In addition, the grade 5 was not separated by standing for 48 hours or more.

From the above description, it can be seen that the above-described embodiments of the present invention achieve the following technical effects:

1) the compound microbial agent prepared by the invention can improve the recovery ratio of oil recovery;

2) the defect that the single-function strain cannot tolerate environmental impact is avoided, and the functional expression among the four strains is not inhibited;

3) the preparation method is simple and low in cost, simplifies the preparation process of the compound microbial agent with the oil extraction function, and effectively improves the oil extraction effect.

The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.