Method for quantifying microorganisms for producing lipopeptide surfactant in microbial flooding reservoir

Abstract

The invention relates to a method for quantifying indigenous microorganisms for producing lipopeptide surfactant in microbial flooding reservoir. Primers are designed according to lipopeptide synthetase genes of microorganisms for producing lipopeptide surfactant, and fluorescent quantitative PCR (polymerase chain reaction) is performed. In optimized reaction procedures and reaction system, the lipopeptide synthetase genes obtained from a standard lipopeptide-producing strain is connected with a carrier to construct a recombinant plasmid used as a standard sample for real-time quantitative PCR; the standard sample is subjected to 10-time gradient dilution and used as a template for fluorescent quantitative PCR, and a standard curve is drawn; a microbial flooding reservoir sample is subjected to simple treatment and used as a template for fluorescent quantitative PCR; and whether data of replicate samples are identical and whether initial templates having different numbers of copies have the same amplification efficiency are judged according to linear relationship R2 and amplification efficiency E. When being used for detecting microorganisms for producing lipopeptide surfactant in reservoir under optimized reaction conditions, the lipopeptide synthetase specific primers have higher stability and sensitivity, thereby being widely used in the field of microbial enhanced oil recovery.

Description

technical field [0001] The invention relates to a quantitative method for microorganisms producing lipopeptide surfactants in oil flooding reservoirs, belonging to the field of microbial oil recovery. Background technique [0002] The production of biosurfactants by microorganisms is one of the main mechanisms for microorganisms to enhance oil recovery. Microbial surfactants mainly include lipopeptides and glycolipids. Lipopeptide is one of the most effective biosurfactants reported so far. Due to its special structure, it can help microbial cells adhere to the surface of hydrocarbons to decompose hydrocarbons, reduce surface tension, and maintain Its activity is of extremely important value to the microbial enhanced oil recovery technology. At present, biosurfactants are produced by microorganisms to enhance oil recovery. There are two methods. One is called surface method, which is to inject biosurfactants produced by microbial surface fermentation into oil reservoirs; t...

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Problems solved by technology

The above two documents respectively use the size of the diameter of the oil discharge circle and the size of the surface tension to characterize the amount of biosurfactant, and then qualitatively reflect the amount of microorganisms producing surfactant. The operation method is simple and easy, but its disadvantage is that it can only qualitatively Judging the presence or amount of surfactant-producing microorganisms, the amount of surfactant-producing microorganisms cannot be quantitatively judged, and there are deficiencies in poor repeatability and low accuracy

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