Method for promoting burkholderia sp. to synthesize toxoflavin by utilizing signal molecules

A technology of Holderella, signaling molecules, applied in the field of microorganisms

Pending Publication Date: 2020-09-15
INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current biosynthetic methods have not been widely used in the synthesis of toxin

Method used

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  • Method for promoting burkholderia sp. to synthesize toxoflavin by utilizing signal molecules
  • Method for promoting burkholderia sp. to synthesize toxoflavin by utilizing signal molecules
  • Method for promoting burkholderia sp. to synthesize toxoflavin by utilizing signal molecules

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Effects of Amino Acids and Mineral Elements on Burkholderia HDXY-02 Toxin Synthesis Ability

[0049] According to the fermentation characteristics and physiological and metabolic characteristics of Burkholderia HDXY-02, amino acids and mineral elements were selected to be added to the medium, and their effects on the synthesis ability of toxin of Burkholderia HDXY-02 were analyzed . The relative value of toxin synthesis in units of bacteria (OD 393 / OD 600 ) to evaluate the toxin synthesis ability of Burkholderia HDXY-02.

[0050] The concentrations of amino acids used are as follows:

[0051] Amino acids: phenylalanine (0.5g / L), tyrosine (0.5g / L), aspartic acid (0.5g / L).

[0052] The selected inorganic salts are as follows: NaCl, KCl, MgSO 4 , CaCl 2 , FeSO 4 , CdCl 2 , MnCl 2 , ZnCl 2 , CuCl 2 .

[0053] The result is as figure 1 As shown, phenylalanine, tyrosine, and aspartic acid cannot significantly improve the toxin synthesis ability of Burk...

Embodiment 2

[0055] Example 2Na 2 SO 4 and K 2 SO 4 Effects on the ability of toxin synthesis in Burkholderia HDXY-02

[0056] NaCl and KCl can improve the toxin synthesis ability and toxin production of Burkholderia HDXY-02, in order to clarify the NaCl + and K + For the promotion of toxin synthesis, choose Na 2 SO 4 and K 2 SO 4 , added to the medium at a certain concentration. Such as Figure 4 As shown, Na 2 SO 4 (85mM-256mM) and K 2 SO 4 (67mM-201mM), like NaCl and KCl, can improve the toxin synthesis ability of Burkholderia HDXY-02, NaCl 2 SO 4 The improvement times are 1.50, 1.54 and 1.39 times of that without adding, K 2 SO 4 The times of improvement were 1.55, 1.39 and 1.34 times when no addition was made. The toxin content in the fermentation supernatant can be increased by 1.28 to 1.63 times respectively. The above results further proved that Na + and K + Promotion of toxin synthesis as a signaling molecule.

Embodiment 3

[0057] Example 3 Other osmotic substances have no promoting effect on the synthetic ability of toxin of Burkholderia HDXY-02

[0058] K + and Na + In addition to being a signaling molecule that can cause osmotic stress, in order to clarify whether the increase in toxin synthesis ability of Burkholderia HDXY-02 is the result of changes in osmotic pressure, different concentrations of manna were added to the medium Alcohol (mannitol), sorbitol (sorbitol) and polyethylene glycol (PEG).

[0059] Permeate substances were used at the following concentrations: mannitol (100 mM, 180 mM, 340 mM), sorbitol (100 mM, 180 mM, 340 mM), PEG (% w / v: 12, 18, 22), NaCl (100 mM, 200 mM, 300 mM). The osmotic pressures corresponding to the three concentrations of each substance are consistent.

[0060] From Figure 5 It can be seen that mannitol, sorbitol, and PEG have no significant effect on the synthesis ability of toxin of Burkholderia HDXY-02, on the contrary, some can inhibit the synthesis...

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Abstract

The invention discloses a method for promoting burkholderia sp. to synthesize toxoflavin by utilizing signal molecules. The signal molecules for promoting the burkholderia to synthesize the toxoflavinare Na<+>, K<+>, Ca<2+> and nitric oxide. The method comprises the following specific steps: carrying out shaking culture on burkholderia sp. HDXY-02 with a preservation number of CGMCC (China General Microbiological Culture Collection Center) N0.14054 overnight at 30 DEG C, and inoculating a sterilization culture medium with the cultured burkholderia sp. HDXY-02 according to an inoculation amount of 1%; and adding one or a combination of two or more of signal molecular compounds (NaCl, Na2SO4, KCl, K2SO4, CaCl2 and nitric oxide donor sodium nitroprusside) with a certain concentration into the culture medium. According to the method, signal molecules are added into the culture solution, so that the capacity of synthesizing the toxoflavin by the burkholderia sp. HDXY-02 and the yield of the toxoflavin are improved to 1.32-3.56 times and 1.28-2.21 times of those of untreated one respectively.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a method for promoting the synthesis of toxoflavin by Burkholderia by using signal molecules. Background technique [0002] Burkholderia (Burkholderia) widely exists in water, soil, plants and human body and other environments, and is a Gram-negative bacterium. Burkholderia can be used in agriculture for biodegradation, biocontrol and promotion of plant growth. Some Burkholderia bacteria can synthesize a small molecule antibacterial substance - toxin. Toxoflavin has been proven to have good inhibitory activity on a variety of tumor cells, and its antifungal activity is good, especially for mutants of Aspergillus fumigatus that can tolerate azoles and Cryptococcus neoformans that can pass through the blood-brain barrier Activity (Xiaodan Li, Yikui Li, Ren Wang, Qizhi Wang, Ling Lu. Toxoflavin produced by Burkholderia gladioli from Lycoris aureais a new broad-spectrum fu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/18C12N1/20C12N1/38C12R1/01
CPCC12P17/182C12N1/20C12N1/38
Inventor 汪仁李晓丹徐晟王蓉周佳宇
Owner INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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