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Method for inhibiting fungal growth by adjusting calcium and iron content and application thereof

A technology for inhibiting the content of fungi and iron, applied in the field of pathogenic fungi growth research, to achieve the effect of enhanced inhibition, good safety and less toxic and side effects

Pending Publication Date: 2022-05-10
NANJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Iron nutrition is necessary for the growth of fungi, and it is also an important element for the survival of pathogens and hosts. Both pathogenic fungi and hosts need iron nutrition. Therefore, a single restriction of iron is obviously not a very effective and safe way to inhibit fungal growth.

Method used

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  • Method for inhibiting fungal growth by adjusting calcium and iron content and application thereof
  • Method for inhibiting fungal growth by adjusting calcium and iron content and application thereof
  • Method for inhibiting fungal growth by adjusting calcium and iron content and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] To prepare Aspergillus fumigatus liquid / solid medium:

[0031] Liquid minimal medium (MM): glucose (Dextrose) 20g / L, trace elements (Trace elements) 1ml / L, sodium nitrate (NaNO 3 ) 6.0g / L, potassium chloride (KCl) 0.52g / L, potassium dihydrogen phosphate (KH 2 PO 4 ) 1.52g / L, magnesium sulfate heptahydrate (MgSO 4 ·7H 2 O) 0.52g / L, uracil (Uracil) 1.2g / L, uridine (Uridine) 1.1g / L. Adjust the pH to 6.5. To prepare solid basic medium, add 2g of agar powder to every 100mL of liquid medium.

[0032] Liquid low-iron plus calcium medium (MM-Fe+Ca): Add 100 μM iron chelator BPS and 10 mM calcium chloride on the basis of removing the trace element ferrous sulfate heptahydrate in the basic medium (MM) formula. If testing other metal ions, replace calcium chloride with corresponding 10mM other metal ions. If preparing a solid low-iron medium, add 2g of agar powder to every 100mL of liquid medium.

[0033] Trace elements: Ferrous sulfate heptahydrate (FeSO 4 ·7H 2 O) 1.0g...

Embodiment 2

[0037] To prepare liquid media for Candida albicans and Cryptococcus neoformans:

[0038] YNB basic medium (YNB): YNB powder (Sigma, Y0626) 6.7g / L, glucose 5g / L.

[0039] YNB low iron medium (YNB-Fe): 20 μM iron chelator BPS was added on the basis of YNB basic medium.

[0040] YNB plus calcium medium (YNB+Ca): Add 100mM exogenous calcium chloride on the basis of YNB basic medium.

[0041] YNB low iron plus calcium medium (YNB-Fe+Ca): 20 μM iron chelator BPS and 100 mM exogenous calcium chloride were added on the basis of YNB basic medium.

[0042] Inoculate 1x10 5 Candida albicans ATCC10231 and Cryptococcus neoformans H99 cells in 100mL liquid YNB basic medium, YNB low iron medium (YNB-Fe), YNB calcium medium (YNB+Ca), YNB low iron calcium medium ( YNB-Fe+Ca), placed in an incubator at 37°C at 200 rpm for 32 hours, and then cultured with shaking at 200 rpm for 32 hours, and the growth was characterized by the absorbance value measured by OD600. Such as figure 2 As shown,...

Embodiment 3

[0044] Establish a nutritional model of mice with low iron and calcium supplementation, and study the ability of calcium and iron combined regulation to inhibit the growth of Aspergillus fumigatus. According to the in vitro culture test, low iron and high calcium have inhibitory effects on the three pathogenic fungi Aspergillus fumigatus, Candida albicans and Cryptococcus neoformans. In the mouse experiment, only Aspergillus fumigatus infection was used as a representative to carry out in vivo experiments.

[0045] Feed 6-week-old C57 mice with iron-deficient feed (Nantong Trofee Feed Technology Co., Ltd., TP0303) with an iron content of only 3 ppm, and gavage 200 μl of calcium chloride with a concentration of 270 mM for 12 consecutive days to establish A mouse model of iron deficiency and calcium supplementation. Inject the immunosuppressant cyclophosphamide to experimental mice, with 2x10 6 Immunosuppressed mice were challenged directly via the trachea with spores of an Asp...

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Abstract

The invention discloses a method for inhibiting fungal growth by adjusting calcium and iron contents and application thereof. The growth inhibition phenomenon of pathogenic fungi is caused by adding exogenous calcium in a low-iron environment, and it is proved that high calcium can inhibit absorption of fungi to iron, so that deletion of iron necessary for fungus growth is aggravated. According to the invention, the iron chelating agent is adopted to reduce the supply of iron nutrition, and calcium nutrition with a certain concentration is combined, so that the growth of aspergillus fumigatus can be obviously inhibited, and the inhibition effect is proved in an in-vitro culture medium and in a mouse body. The invention further finds that high-calcium and low-iron combined regulation can also inhibit pathogenic fungi-candida albicans and cryptococcus neoformans. The low-iron and high-calcium combined nutrition treatment technology disclosed by the invention can be popularized to control the growth of other types of pathogenic fungi such as plants, animals and the like, and is also effective to antifungal drug resistant strains.

Description

technical field [0001] The invention belongs to the field of research on the growth of pathogenic fungi, and in particular relates to a method for regulating calcium and iron content to inhibit fungal growth and an application thereof. Background technique [0002] Pathogenic fungi have a huge impact on humans, animals, plants and ecosystems, and understanding the survival mechanisms of fungal pathogens in different host niches is very important for controlling the invasion of fungal pathogens. Iron is an essential micronutrient involved in many important cellular biological processes as a cofactor for several enzymes, including oxidoreductases. In order to overcome the low bioavailability of iron during the invasion of the host, pathogens have evolved a variety of sophisticated systems to compete with the host for iron, control their own iron absorption, consumption and storage, and maintain iron homeostasis. Process matters. [0003] The commonly used methods for inhibit...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N1/16A01N43/90A01N37/20A01N59/06A01P3/00A61K31/164A61K31/4745A61K33/14A61P31/10C12R1/68C12R1/725C12R1/645
CPCC12N1/14C12N1/16A01N43/90A01N37/20A01N59/06A61K31/164A61K31/4745A61K33/14A61P31/10A61K2300/00
Inventor 陆玲张元炜叶婧万沁怡
Owner NANJING NORMAL UNIVERSITY
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