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Secretions of marine halomonas and extraction method and application of secretions

A Halomonas, extraction method technology, applied in the directions of botanical equipment and methods, microorganism-based methods, biochemical equipment and methods, etc. Fast action time, simple culture conditions, and good instant inhibition effect

Inactive Publication Date: 2015-04-29
SHANTOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the sea, adding algae-inhibiting substances to inhibit algae species must have an instant algicidal effect. If the effect lasts for a long time, it will be diluted quickly due to the dilution of seawater. Therefore, no effective anti-red tide has been found so far. algae matter
Even if some substances have this effect, they cannot be applied in practice due to the cumbersome extraction process and high cost

Method used

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  • Secretions of marine halomonas and extraction method and application of secretions
  • Secretions of marine halomonas and extraction method and application of secretions
  • Secretions of marine halomonas and extraction method and application of secretions

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The specific process of extracting the secretion of Halomonas marine in the present embodiment is as follows: figure 1 shown.

[0029] Step 1, prepare the medium according to the following ratio: 5g of yeast powder, 10g of tryptone and 1L of seawater;

[0030] Step 2: Sterilize the prepared medium at 121°C for 15 minutes;

[0031] Step three, will be as figure 2 The shown Halomonas marine bacteria were inoculated into the culture medium under sterile conditions, and cultured on a shaker at a constant temperature of 120 rpm at 30°C for 10 days, and the bacteria could fully secrete metabolites;

[0032] Step 4, use a 0.45 μm membrane to filter and sterilize to obtain the bacterial secretion supernatant, OD 267 is 1.4;

[0033] Step 5, the obtained filtrate is vacuum freeze-dried, then extracted with pure acetic acid, and desalted by filter paper filtration;

[0034] Step 6, after evaporating the obtained filtrate to dry acetic acid, the bold substance of the secreti...

Embodiment 2

[0037] The specific process of extracting the secretion of Halomonas marine in the present embodiment is as follows: figure 1 shown.

[0038] Step 1, prepare the medium according to the following ratio: 5g of yeast powder, 10g of tryptone and 1L of seawater;

[0039] Step 2: Sterilize the prepared culture medium at 121°C for 20 minutes;

[0040] Step three, will be as figure 2 The shown Halomonas marine bacteria were inoculated into the culture medium under sterile conditions, and cultured on a shaker at 25°C and 150 rpm for 15 days, and the bacteria could fully secrete metabolites;

[0041] Step 4, use a 0.22 μm membrane to filter and sterilize to obtain the bacterial secretion supernatant, OD 267 is 1.6;

[0042] Step 5, the obtained filtrate is vacuum freeze-dried, then extracted with pure acetic acid, and desalted by filter paper filtration;

[0043] Step 6, after evaporating the obtained filtrate to dry acetic acid, the bold substance of the secretion of Halomonas m...

Embodiment 3

[0045] The crude extract and the algae liquid were mixed and cultivated according to the ratio of 1:50, 1:100, 1:200, 1:500, cultured for 8 days, and the concentration was 14.0mg / 100mL, 7.0mg / 100mL, 3.5mg / 100mL, 1.4mg / 100mL secretion of Halomonas marine. Bold objects used to study the effects of different concentrations of secretions on the inhibition of different algae (blank control).

[0046] image 3 The pairs in bold that are the secretions of Halomonas marine bacteria at different concentrations are the cases of inhibition by green algae, diatoms and cyanobacteria, where green algae include Chlorella, diatoms include Phaeodactylum tricornutum, Chaetoceros gracilis, Nitzschia minor and Skeletons costata, cyanobacteria including Cylindrocystis and Microcystis aeruginosa, their initial OD 680 About 0.04 (these cells are relatively small, the absorbance is used as the size of the algae cell concentration); experiments show that after cultivating for 8 days, the secretion ...

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Abstract

The invention relates to secretions of marine halomonas, an extraction method of the secretions and the application of the secretions. The extraction method comprises the following steps: preparing a culture medium according to a formula of 5 g of yeast powder, 10 g of tryptone and 1L of seawater, and sterilizing the culture medium for 15-20 minutes at the temperature of 121 DEG C; inoculating the marine halomonas into the culture medium under aseptic conditions, culturing the culture medium inoculated with the marine halomonas for 10-15 days at the constant temperature of 25-30 DEG C on a backward rotating shaking table at the speed of 120-150 rpm, performing filtration with a film of 0.22-0.45 micrometers, and performing sterilization so as to obtain bacterial exudate supernatant; vacuum freezing and drying a filtrate, performing extraction through a pure acetic acid, and performing filtration and desalination through filter paper; volatilizing the acetic acid of the obtained filtrate so as to obtain coarse substances of the secretions of the marine halomonas. The secretions disclosed by the invention can be used for governing red tides, and have the effect of restraining dinoflagellates like Alexandrium tamarenses, prorocentrum donghaiense and Karenia mikimotoi of the red tides. According to the invention, the culture conditions are simple, products do not need to be refined, only the filtration is needed for removing thalli, and the secretions have a good effect of restraining the dinoflagellates of the red tides.

Description

technical field [0001] The present invention relates to the secretion of marine halomonas and its extraction method and application. Background technique [0002] In recent years, there have been frequent outbreaks of dinoflagellate blooms in the ocean, and most red tide dinoflagellates produce toxins, such as hemolysin, paralytic shellfish poisoning (PSP), brevetoxin- group, BTX), not only the red tide itself has serious consequences on the marine ecological environment, but also these toxins produced by algae will cause great harm to marine animals such as fish and shellfish, and even threaten people's health, so they are highly valued by coastal countries . Judging from the existing conditions, once a large area of ​​red tide occurs, there is no particularly effective method to stop it. For some local and small-scale methods of preventing and controlling red tide, although many experiments have been done, the effect is not ideal enough. Mainly use chemical drugs (copper...

Claims

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Application Information

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IPC IPC(8): C12P1/04A01N63/02A01P13/00C12R1/01
Inventor 朱兴彪谢丽玲郑玉忠黄文浩张振霞朱炎坤李培培
Owner SHANTOU UNIV
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