阅读说明：本技术 油藏石油烃降解菌的激活剂的筛选及激活效果评价方法 (Method for screening activating agent of oil reservoir petroleum hydrocarbon degrading bacteria and evaluating activating effect ) 是由 李海兰 张忠智 孙珊珊 来瑞秋 姚甜甜 龚页境 张慧慧 张沛琳 杨宁 于 2019-09-18 设计创作，主要内容包括：本发明提供了一种油藏石油烃降解菌的激活剂的筛选及激活效果评价方法。本发明的筛选方法包括：采用单因素法筛选氮源和/或磷源种类；采用油藏地层水配制待选激活剂,所述激活剂包括碳源、氮源、磷源、硫酸镁、氯化钙、酵母粉,限氧培养,100～200rpm在目标油藏温度振荡培养2d～4d,得到激活后的发酵液；取激活后的发酵液进行OD600、乳化能力和/或表面张力测定,根据测定结果筛选具有定向调控油藏石油烃降解功能菌功效的激活剂。本发明筛选得到的激活剂具有高效、成本低廉、定向激活、绿色环保、稳定传代等特点,有助于进一步提高原油采收率。(The invention provides a method for screening an activator of oil reservoir petroleum hydrocarbon degrading bacteria and evaluating the activation effect. The screening method of the invention comprises the following steps: adopting a single-factor method to screen the types of nitrogen sources and/or phosphorus sources; preparing an activator to be selected by using oil reservoir formation water, wherein the activator comprises a carbon source, a nitrogen source, a phosphorus source, magnesium sulfate, calcium chloride and yeast powder, carrying out oxygen-limited culture, and carrying out shake culture at 100-200 rpm at a target oil reservoir temperature for 2-4 d to obtain activated fermentation liquor; and (3) measuring OD600, emulsifying capacity and/or surface tension of the activated fermentation liquor, and screening an activator with the effect of directionally regulating and controlling the degradation function bacteria of the petroleum hydrocarbon in the oil reservoir according to the measurement result. The activator obtained by screening has the characteristics of high efficiency, low cost, directional activation, environmental protection, stable passage and the like, and is favorable for further improving the crude oil recovery ratio.)

1. A screening method of an activator for directionally activating petroleum hydrocarbon degradation functional bacteria comprises the following steps:

adopting a single-factor method to screen the types of nitrogen sources and/or phosphorus sources;

preparing an activator to be selected by using oil reservoir formation water, wherein the activator comprises a carbon source, a nitrogen source, a phosphorus source, magnesium sulfate, calcium chloride and yeast powder, carrying out oxygen-limited culture, and carrying out shake culture at 100-200 rpm at a target oil reservoir temperature for 2-4 d to obtain activated fermentation liquor;

and (3) measuring OD600, emulsifying capacity and/or surface tension of the activated fermentation liquor, and screening an activator with the effect of directionally regulating and controlling the degradation function bacteria of the petroleum hydrocarbon in the oil reservoir according to the measurement result.

2. The method of claim 1, further comprising:

the activating agent is subjected to directional activation of petroleum hydrocarbon degradation functional bacteria, the activation effect, the action with crude oil after activation and the stability in the activation process are evaluated, and therefore the activating agent is evaluated and screened.

3. The method according to claim 1, wherein the candidate activators comprise a carbon source, a nitrogen source, a phosphorus source, magnesium sulfate, calcium chloride and yeast powder, and the mass ratio of the components is as follows: 1-4 parts of carbon source, 0.17-1.0 part of nitrogen source, 0.4-1.4 parts of phosphorus source, 0.03-0.08 part of magnesium sulfate, 0.003-0.008 part of calcium chloride and 0.03-0.08 part of yeast powder;

preferably, in an activator system prepared by using oil reservoir formation water, the mass fraction of a mixed system is calculated by 100%, wherein the mass fraction of a carbon source is 1-4% of the carbon source.

4. The method of claim 1, further comprising measuring the total number of bacteria, hydrocarbon degrading bacteria, and sulfate reducing bacteria in the activated fermentation broth.

5. The method of claim 2, wherein evaluating the effect on the crude oil after activation comprises observing changes in the saturated hydrocarbon composition and oil droplet size of the crude oil.

6. The method of claim 2, wherein evaluating stability during activation comprises: taking the activated culture solution for transfer; the new transfer culture medium is prepared by deionized water, 5-10% of activated culture solution is added, and the mass concentrations of carbon source, nitrogen source, phosphorus source, magnesium sulfate, calcium chloride and yeast powder are respectively 1-4%, 0.17-1.0%, 0.4-1.4%, 0.03-0.08%, 0.003-0.008% and 0.03-0.08%; limiting oxygen, carrying out shake culture at the target oil reservoir temperature for 2 d-4 d at 100-200 rpm, continuously carrying out passage, and determining the structure composition of the biological community.

7. The method of claim 2, comprising evaluating the targeted activation effect of the petroleum hydrocarbon degrading functional bacteria in the reservoir according to the rules shown in the following table:

in the table, the comprehensive scores of the directional activators of the oil reservoir petroleum hydrocarbon degradation functional bacteria are calculated, wherein the comprehensive scores are more than 80 and belong to the I class, 70-79 belong to the II class, and 50-69 belong to the III class.

8. An activator for directionally activating petroleum hydrocarbon degradation functional bacteria comprises a carbon source, a nitrogen source, a phosphorus source, magnesium sulfate, calcium chloride and yeast powder, wherein the mass ratio of the components is as follows: 1-4 carbon sources, 0.17-1.0 nitrogen source, 0.4-1.4 phosphorus source, 0.03-0.08 magnesium sulfate, 0.003-0.008 calcium chloride and 0.03-0.08 yeast powder.

9. The activator of claim 8, wherein the carbon source is reservoir crude oil, the nitrogen source is a combination of sodium nitrate and ammonium sulfate, and the phosphorus source is a combination of potassium dihydrogen phosphate and sodium dihydrogen phosphate;

preferably, the nitrogen source is a compound of sodium nitrate and ammonium sulfate according to the mass ratio of 1: 1-5: 1, and the phosphorus source is a compound of potassium dihydrogen phosphate and sodium dihydrogen phosphate according to the mass ratio of 5: 1-1: 1.

10. Use of the activator according to claim 8 or 9 for the targeted activation of petroleum hydrocarbon degrading functional bacteria and/or enhanced oil recovery.

Technical Field

The invention relates to a method for screening an activator of oil deposit petroleum hydrocarbon degrading bacteria and evaluating the activation effect, in particular to a method for screening a formula of the activator of the oil deposit petroleum hydrocarbon degrading bacteria with directional regulation and control functions and evaluating the activation effect, and a method for directionally activating the petroleum hydrocarbon degrading bacteria by using an activator system obtained by screening so as to be applied to oil displacement, belonging to the technical field of microbial oil displacement.

Background

Most oil reservoirs contain abundant microbial resources, and the unique oil reservoir environment creates unique microbial resources because the microbial community structure is changed due to slight change of the environment under the extreme oil reservoir environment. Practice shows that the microbial oil production, especially the endogenous microbial oil production technology, is a technology with relatively high application potential, and compared with chemical flooding, polymer flooding and other technologies, the technology has the advantages of low cost, environmental friendliness, strong sustainability and no fluctuation of the crude oil price.

An endogenous microbial community is one that remains relatively stable in number and type over a period of time during oilfield waterflood development. The microorganism in the oil deposit is a complex community, and the microbial oil production technology is actually used for directionally regulating and controlling the microorganism community, limiting and inhibiting the number of non-functional bacteria, increasing and playing the number and the function of functional microorganism populations so as to enable the microorganisms to become a community which is beneficial to oil production. Some microorganisms enter the oil reservoir with the injection of water, and waterflooding is a major factor in determining the distribution of endogenous microbial communities in the reservoir ecosystem. In deeper reservoirs the structure of the endogenous microbial community is relatively simple due to the high temperature and pressure; in a shallow stratum, due to low temperature and pressure, the endogenous microbial community is complex, has multiple types and large quantity, mainly comprises bacteria, and mainly comprises Hydrocarbon Oxidizing Bacteria (HOB), zymocyte (FB), Methanogen (MPB), Sulfate Reducing Bacteria (SRB), Nitrate Reducing Bacteria (NRB), iron bacteria, sulfur bacteria and the like. During the long-term survival process of the bacteria in the specific environment of the oil reservoir, some bacteria die without adapting to the environment of the oil reservoir, such as some surface mixed bacteria; some bacteria are in dormant state, such as Bacillus spp; some bacteria can survive in the specific environment of oil reservoir through self function regulation and utilize limited nutritive matter in oil reservoir stratum for slow growth, propagation and metabolism, such as pseudomonads, etc.

In summary, in the prior art, the oil reservoir petroleum hydrocarbon degrading bacteria have low growth and metabolic capability, and the oil recovery functional bacteria have low survival number, action period, thallus activity, product amount and the like in the whole oil reservoir microbial metabolic chain. In terms of enlarging the application scale of the technology, the injection mode still has high cost, and the cost control is a key point of long-term attack of the microbial oil recovery technology.

Disclosure of Invention

The invention aims to provide a method for screening an activator of oil reservoir petroleum hydrocarbon degrading bacteria and evaluating the activation effect, so as to screen the activator which has the characteristics of high efficiency, low cost and the like and has the function of activating the oil reservoir endogenous petroleum hydrocarbon degrading bacteria.

In one aspect, the invention provides a screening method of an activator for directionally activating petroleum hydrocarbon degradation functional bacteria, which comprises the following steps:

adopting a single-factor method to screen the types of nitrogen sources and/or phosphorus sources;

preparing an activator to be selected by using oil reservoir formation water, wherein the activator comprises a carbon source, a nitrogen source, a phosphorus source, magnesium sulfate, calcium chloride and yeast powder, carrying out oxygen-limited culture, and carrying out shake culture at 100-200 rpm at a target oil reservoir temperature for 2-4 d to obtain activated fermentation liquor;

and (3) measuring OD600, emulsifying capacity and/or surface tension of the activated fermentation liquor, and screening an activator with the effect of directionally regulating and controlling the degradation function bacteria of the petroleum hydrocarbon in the oil reservoir according to the measurement result.

According to the specific embodiment of the invention, the screening method of the activating agent for directionally activating the petroleum hydrocarbon degradation functional bacteria further comprises the following steps:

the activating agent is subjected to directional activation of petroleum hydrocarbon degradation functional bacteria, the activation effect, the action with crude oil after activation and the stability in the activation process are evaluated, and therefore the activating agent is evaluated and screened.

According to the specific embodiment of the invention, in the screening method of the activator for directionally activating the petroleum hydrocarbon degradation functional bacteria, the to-be-selected activator comprises a carbon source, a nitrogen source, a phosphorus source, magnesium sulfate, calcium chloride and yeast powder, and the mass ratio of the components is as follows: 1-4 carbon sources, 0.17-1.0 nitrogen source, 0.4-1.4 phosphorus source, 0.03-0.08 magnesium sulfate, 0.003-0.008 calcium chloride and 0.03-0.08 yeast powder. Preferably, in an activator system prepared by using oil reservoir formation water, the mass fraction of a mixed system is calculated by 100%, wherein the mass fraction of a carbon source is 1-4% of the carbon source.

According to the specific embodiment of the invention, the screening method of the activating agent for directionally activating the petroleum hydrocarbon degradation functional bacteria further comprises the process of measuring the total bacteria number, the hydrocarbon degradation bacteria and the sulfate reduction bacteria by taking the activated fermentation liquor.

According to the specific embodiment of the invention, in the screening method of the activating agent for directionally activating the petroleum hydrocarbon degradation functional bacteria, the evaluation of the action of the activated activating agent and the crude oil comprises the observation of the composition change of the saturated hydrocarbon of the crude oil and the grain size of oil drops.

According to a specific embodiment of the invention, in the screening method of the activating agent for directionally activating the petroleum hydrocarbon degradation functional bacteria, the evaluation of the stability in the activating process comprises the following steps: taking the activated culture solution for transfer; the new transfer culture medium is prepared by deionized water, 5-10% of activated culture solution is added, and the mass concentrations of carbon source, nitrogen source, phosphorus source, magnesium sulfate, calcium chloride and yeast powder are respectively 1-4%, 0.17-1.0%, 0.4-1.4%, 0.03-0.08%, 0.003-0.008% and 0.03-0.08%; limiting oxygen, carrying out shake culture at the target oil reservoir temperature for 2 d-4 d at 100-200 rpm, continuously carrying out passage, and determining the structure composition of the biological community.

According to the specific embodiment of the invention, the screening method of the activating agent for directionally activating the petroleum hydrocarbon degradation functional bacteria comprises the following steps of evaluating the directional activation effect of the petroleum hydrocarbon degradation functional bacteria according to the rules shown in the following table 1:

TABLE 1 evaluation rule for directional activation effect of oil reservoir petroleum hydrocarbon degradation functional bacteria

In the table, the comprehensive scores of the directional activators of the oil reservoir petroleum hydrocarbon degradation functional bacteria are calculated, wherein the comprehensive scores are more than 80 and belong to the I class, 70-79 belong to the II class, and 50-69 belong to the III class.

On the other hand, the invention also provides an activator for directionally activating the petroleum hydrocarbon degradation functional bacteria, which comprises a carbon source, a nitrogen source, a phosphorus source, magnesium sulfate, calcium chloride and yeast powder, wherein the mass ratio of the components is as follows: 1-4 carbon sources, 0.17-1.0 nitrogen source, 0.4-1.4 phosphorus source, 0.03-0.08 magnesium sulfate, 0.003-0.008 calcium chloride and 0.03-0.08 yeast powder.

According to the specific embodiment of the invention, in the activator, the carbon source is crude oil in an oil reservoir, the nitrogen source is a compound of sodium nitrate and ammonium sulfate, and the phosphorus source is a compound of potassium dihydrogen phosphate and sodium dihydrogen phosphate. Preferably, the nitrogen source is a compound of sodium nitrate and ammonium sulfate according to the mass ratio of 1: 1-5: 1, and the phosphorus source is a compound of potassium dihydrogen phosphate and sodium dihydrogen phosphate according to the mass ratio of 5: 1-1: 1.

The invention also provides application of the activator in directionally activating the petroleum hydrocarbon degradation functional bacteria.

The invention also provides application of the activator in improving the recovery ratio of the oil reservoir.

The invention mainly evaluates the activating characteristic, the emulsifying effect and the stable passage of the oil deposit petroleum hydrocarbon degradation functional bacteria and has the function of directionally regulating and controlling the petroleum hydrocarbon degradation functional bacteria activating agent. The method can effectively evaluate the propagation metabolism condition and the emulsification capacity change of the oil deposit petroleum hydrocarbon degradation functional bacteria, and can evaluate the stable passage of the activated petroleum hydrocarbon degradation functional bacteria, thereby enhancing the stability of the petroleum hydrocarbon degradation functional bacteria in the microbial oil displacement technology for further expanding the application.

The activating agent obtained by screening has the characteristics of high efficiency, low cost, directional activation, environmental protection, stable passage and the like, heavy components in the crude oil are degraded by activating endogenous microorganisms in the oil reservoir to reduce the viscosity of the crude oil, enhance the flow capacity of the crude oil, or metabolize to generate products such as a biosurfactant and the like, the interfacial tension of the crude oil and the surface of rock of the oil reservoir is reduced, dominant bacteria in oil reservoir petroleum hydrocarbon degradation functional bacteria are converted into bacillus subtilis with good oil displacement effect, the passage is stable, and the recovery ratio of the crude oil is further improved.

Drawings

FIG. 1 is a schematic diagram of an evaluation method of an anaerobic inorganic salt activation system of a directional activator of oil reservoir petroleum hydrocarbon degradation functional bacteria.

FIG. 2 is a schematic diagram of an evaluation method of an aerobic-oxygen limited system of a directional activator of petroleum hydrocarbon degradation functional bacteria in an oil reservoir.

FIG. 3 is a schematic diagram of an evaluation method of an oil reservoir petroleum hydrocarbon degradation functional bacteria directional activator oxygen limiting system.

FIG. 4 is a photograph of the particle size distribution of crude oil before and after the action of the activator.

Fig. 5A and 5B are graphs showing the change in the composition of normal paraffins and naphthenes in the crude oil saturates before and after activation.

Detailed Description

The following examples are intended to illustrate the practice and advantageous effects of the present invention, but are not to be construed as limiting the scope of the present invention. The procedures and operating conditions not specified in detail in the examples were carried out according to the usual procedures in the art.

Whether the oil is removed by endogenous microorganisms or exogenous microorganisms, the microorganisms play a role in oil displacement only by the existence of a certain number of functional microorganism populations through 'winning by quantity'. The invention optimizes the formula by inspecting the number of the petroleum hydrocarbon degrading bacteria activated by endogenous microorganisms and inspects the stable passage of the petroleum hydrocarbon degrading bacteria.