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Method for preparing ciguatoxin by utilizing fermenting of marine toxin-producing dinoflagellate symbiotic bacteria

A technology of xiga toxin and bacterial fermentation, applied in the field of marine microorganism fermentation, can solve the problems of high cost and long cultivation period, and achieve the effects of low cost, short period and small workload

Active Publication Date: 2018-12-11
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method not only has a long cultivation cycle and high cost, but also requires special algae cultivation equipment

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] (1) Shake flask seed solution: Inoculate the marine dinoflagellate symbiotic bacterial strain (Haliea sp.) LZ16-2 preserved on a solid slope into a 1L volumetric Erlenmeyer flask with a liquid volume of 200mL, and incubate at 28°C for 12 hours. Light microscope observation shows that the growth of the bacteria is good, and there is no contamination by bacteria, so it can be inoculated into the seed tank;

[0027] (2) Preparation of seed solution: Prepare 1L of culture solution according to the culture medium formula of the shake flask seed solution, add it to a 5L volume seed tank, heat it with steam, sterilize at 121°C for 30 minutes, cool to 28°C, and inoculate with flame method. The shake flask seed solution is inoculated on the seed medium in the seed tank. The composition of the seed medium is: 5g of yeast extract, 4g of peptone, 1g of glucose and 1000mL of natural seawater; the pH of the seed medium is 6.8; the culture conditions are: ventilation 50L / min, stirring...

Embodiment 2

[0032] (1) Shake flask seed solution: Inoculate the marine dinoflagellate symbiotic bacterial strain (Haliea sp.) LZ16-2 preserved on a solid slope into a 1L volumetric Erlenmeyer flask with a liquid volume of 200mL, and incubate at 28°C for 12 hours. Light microscope observation shows that the growth of the bacteria is good, and there is no contamination by bacteria, so it can be inoculated into the seed tank;

[0033] (2) Preparation of seed solution: Prepare 1L of culture solution according to the culture medium formula of the shake flask seed solution, add it to a 5L volume seed tank, heat it with steam, sterilize at 121°C for 30 minutes, cool to 28°C, and inoculate with flame method. The shake flask seed solution is inoculated on the seed medium in the seed tank. The composition of the seed medium is: 5g of yeast extract, 4g of peptone, 1g of glucose and 1000mL of natural seawater; the pH of the seed medium is 6.8; the culture conditions are: ventilation 60L / min, stirring...

Embodiment 3

[0038] (1) Shake flask seed solution: Inoculate the marine dinoflagellate symbiotic bacterial strain (Haliea sp.) LZ16-2 preserved on a solid slope into a 1L volumetric Erlenmeyer flask with a liquid volume of 200mL, and incubate at 28°C for 12 hours. Light microscope observation shows that the growth of the bacteria is good, and there is no contamination by bacteria, so it can be inoculated into the seed tank;

[0039] (2) Preparation of seed solution: Prepare 1L of culture solution according to the culture medium formula of the shake flask seed solution, add it to a 5L volume seed tank, heat it with steam, sterilize at 121°C for 30 minutes, cool to 28°C, and inoculate with flame method. The shake flask seed solution is inoculated on the seed medium in the seed tank. The composition of the seed medium is: 5g of yeast extract, 4g of peptone, 1g of glucose and 1000mL of natural seawater; the pH of the seed medium is 6.8; the culture conditions are: ventilation 55L / min, stirring s...

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Abstract

The invention relates to the technical field of marine microorganism fermentation, in particular to a method for preparing ciguatoxin by utilizing fermenting of marine toxin-producing dinoflagellate symbiotic bacteria. The method comprises the following steps of (1) shaking of a seed liquid bottle; (2) preparation of a seed liquid; (3) fermenting and culture; (4) extracting and purifying of the fermenting liquid; (5) purifying of crude extract: eluting by preparation of liquid phase chromatography, collecting, rotating and evaporating, so as to obtain a white powder, namely the ciguatoxin. Themethod has the advantages that the marine dinoflagellate symbiotic bacteria straini Haliea sp.LZ16-2 is used as a production bacterial strain; by utilizing the bacterial strain to ferment, the yieldrate of ciguatoxin is higher, and can reach 21.1 to 25.3mug / L, the fermenting time is 60 to 64h, and the energy consumption is decreased; the steps are simple, the workload is small, the cycle is shorter, the used reagents and materials are common reagents for microorganism fermenting test, the injury to the human body is avoided, the environment-friendly effect is realized, the cost is low, and the higher market prospect and economic value are realized.

Description

technical field [0001] The invention relates to the technical field of marine microbial fermentation, in particular to a method for preparing sija toxin by fermenting marine toxin-producing dinoflagellate symbiotic bacteria. Background technique [0002] Ciguatoxin (CTX) is a heat-stable macromolecular polyether neurotoxin with strong toxicity, more than 100 times stronger than tetrodotoxin, and is one of the more serious red tide biotoxins known. CTX is a voltage-dependent Na + Channel agonists have important effects on the nervous system, digestive system and cardiovascular system. It can be used as a molecular tool to study the structure and function of excited cell membranes and the contraction mechanism of local anesthetics. In addition, CTX has important applications in the detection of harmful red tides, neurobiology, medical diagnosis, and biochemical warfare agents. [0003] At present, sijatoxin is mainly obtained through large-scale cultivation of dinoflagellat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/18C12R1/01
CPCC12P17/181
Inventor 杨桥张晓玲穆军蒋志伟张若男
Owner ZHEJIANG OCEAN UNIV
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