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Method for rapidly determining content of polyunsaturated fatty acids in oil-containing microorganism cells

A technology for unsaturated fatty acids and microbial cells, which is applied in the field of quickly determining the content of unsaturated fatty acids in cells and the content of polyunsaturated fatty acids in cells. The effects of manpower and material resources, sample preparation and simple detection process

Active Publication Date: 2015-09-30
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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Problems solved by technology

The samples required by these methods usually need to be prepared through complicated steps such as solvent extraction and derivatization, which not only takes a long time, but also may affect the detection accuracy and cause large errors

Method used

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  • Method for rapidly determining content of polyunsaturated fatty acids in oil-containing microorganism cells
  • Method for rapidly determining content of polyunsaturated fatty acids in oil-containing microorganism cells
  • Method for rapidly determining content of polyunsaturated fatty acids in oil-containing microorganism cells

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Embodiment 1

[0023] The present embodiment is to use the method of the present invention to compare a kind of microalgae, the polyunsaturated fatty acid content of Schizochytrium sp. under different culture conditions, and concrete steps are as follows:

[0024] 1. Sample preparation: take 1 ml each of 9 Schizochytrium cell samples under different culture conditions and centrifuge at a speed of 8000 g to remove the supernatant; in phosphate buffer. The bacterial suspension was centrifuged at a speed of 8000 g to remove the supernatant; the bacterial pellet was resuspended in phosphate buffer equal to the volume of the bacterial cells. The phosphate buffer used in sample preparation has the same pH and ionic strength as the culture medium.

[0025] 2. Nuclear magnetic resonance detection: take 500 microliters of cell resuspension, add 50 microliters of heavy water, place it in a nuclear magnetic resonance sample tube with an outer diameter of 5 mm, and measure it on a 600 MHz nuclear magne...

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Abstract

The invention discloses a method for rapidly determining the content of polyunsaturated fatty acids in oil-containing microorganism cells. The method is used for mainly detecting the relative content of polyunsaturated fatty acids in the cells in microorganisms including microalgae, bacteria, fungi, yeast and the like; after a cell culture solution is centrifuged, the bacteria are re-suspended in a buffering solution, and a nuclear magnetic resonance 1H spectrum or 13C spectrum is determined; and the contents of polyunsaturated fatty acids of the different samples can be compared according to the strength ratio of a characteristic peak on a nuclear magnetic resonance spectrogram. Compared with a traditional fatty acid content determination method, operations including sample drying, cell crushing, grease extraction, transesterification and the like are not needed and an organic solvent is not needed, so that manpower and material resources are saved and the determination efficiency is greatly improved; and the method is suitable for high-throughput rapid detection of a lot of samples, such as microorganism screening, optimization of culture conditions and monitoring of an industrial production process.

Description

technical field [0001] The invention belongs to the technical field of detection methods, in particular to a method for the content of polyunsaturated fatty acids in cells, and in particular to a rapid determination suitable for strain screening, optimization of microbial culture conditions, and monitoring of microbial culture processes for industrial production of polyunsaturated fatty acids Methods for intracellular unsaturated fatty acid content. Background technique [0002] Polyunsaturated fatty acids (mainly including DHA, EPA, ARA, etc.) have many important physiological functions such as maintaining cell homeostasis and normal growth, preventing cardiovascular diseases, immune regulation, and anticancer. Polyunsaturated fatty acids have long been used as important additives in infant milk powder, health food, and medicines. In recent years, they have also begun to be used as additives in edible oils, dairy products and other foods to improve their nutritional value. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N24/08
Inventor 冯银刚兰君谭延振宋晓金崔球
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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