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Method for extracting mycosporine-like amino acid shinorine from microcystis

A microcystis and amino acid technology, which is applied in the fields of biotechnology, resource utilization and sustainable development, can solve the problems of no mycosporin-like amino acids, human skin is easy to cause damage, and is not conducive to human health, so that the extract has less impurities, The effect of convenient operation and simple equipment

Active Publication Date: 2014-04-30
INST OF AQUATIC LIFE ACAD SINICA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the photoprotective substances used in my country are mainly chemically synthesized. Long-term use of chemically synthesized substances is likely to cause damage to human skin, which is not conducive to human health.
[0004] Moreover, the current domestic extraction of mycosporine-like amino acids is rarely involved, and the purity is not high
Especially mycosporin-like amino acid standard products that are not commercially available in the world at present

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Culture of Microcystis:

[0034] 1. Preparation of culture medium: conventional cyanobacteria culture medium;

[0035] 2. Culture of Microcystis:

[0036] (1) Primary culture: Inoculate the original seed in the test tube (4ml) into a 250ml culture bottle (with 200ml of medium in it), culture with ventilation, adjust the airflow to 1-2L / min, and pass the airflow through a needle filter (0.22μm ) aseptic treatment connection, the light intensity is controlled at 40-60μEm -2 the s -1 , the temperature is controlled at 25-28°C, and after 5-8 days of cultivation, transfer to secondary cultivation;

[0037] (2) Secondary culture: Put all the cultures obtained from the first-level culture into a 2L culture bottle (with 1L of culture medium inside), ventilate the culture, adjust the airflow to 3-4L / min, and pass the airflow through the needle filter ( 0.22μm) aseptic connection, the light intensity is controlled at 60-80μEm -2 the s -1 , the temperature is controlled at 2...

Embodiment 2

[0042] A method for extracting and preparing mycosporine-like amino acid Shinorine from Microcystis, the steps are as follows:

[0043] 1. Weigh 10g of Microcystis powder prepared in Example 1 and add 100ml of anhydrous methanol, first place it in an ultrasonic cleaner for 100% ultrasonic treatment for 15 minutes, then place it in a water bath at 45°C for 3 hours, and mix it up and down during the process. Homogenize 10 times, centrifuge at 11000rpm / min for 15min, take the supernatant, add 100ml of anhydrous methanol to the filter residue for extraction, repeat the extraction 3 times, and combine the collected supernatants;

[0044] 2. Place the collected supernatant in a nitrogen blower, dry it with nitrogen, and redissolve it with 10ml of distilled water;

[0045] 3. Take out the reconstituted solution, add an equal volume of chloroform, and mix well;

[0046] 4. Centrifuge at 11000rpm / min for 15min to collect the supernatant;

[0047] 5. Pass the supernatant through a 0.2...

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Abstract

The invention discloses a method for extracting mycosporine-like amino acid shinorine from microcystis. The method particularly comprises the following steps: (1) mixing microcystis powder with methanol, treating by ultrasounds and heating, centrifuging and collecting supernatant; (2) blowing the supernatant by using nitrogen to be dried and re-dissolving with distilled water; (3) adding chloroform and uniformly mixing; (4) centrifuging and collecting supernatant; (5) filtering by using a needle type filter; (6) treating by using a strata-x solid-phase extraction small column, and freeze-drying filtrate. The method for extracting the mycosporine-like amino acid shinorine is simple, convenient to operate and safe and reliable; the purity of a final product is up to 85%-95% and the final product can be used as a standard product and lays a firm foundation for follow-up large-scale quantification.

Description

technical field [0001] The invention relates to the fields of biotechnology, resource utilization and sustainable development, and more specifically relates to a method for extracting mycosporine-like amino acid Shinorine from Microcystis. Background technique [0002] At present, the utilization of Microcystis is almost blank, let alone the extraction and utilization of mycosporin-like amino acids from Microcystis. [0003] Mycosporine and mycosporine-like amino acids (mycosporine and mycosporine-like) are a large class of water-soluble substances formed by condensation with different types of amino acids with cyclohexenone as the basic skeleton. They can be used in fungi, marine bacteria, cyanobacteria and eukaryotic algae. Synthesized in cells and accumulated in organisms such as marine invertebrates and fish, its molecular size is generally less than 400Da, and this type of substance has a strong light absorption capacity in the range of 310-360nm. Many Microcystis cont...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C251/20C07C249/02
Inventor 高合意虞功亮李仁辉
Owner INST OF AQUATIC LIFE ACAD SINICA
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