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CN-121991812-A - Haemophilus with high yield of active spores and culture method thereof

CN121991812ACN 121991812 ACN121991812 ACN 121991812ACN-121991812-A

Abstract

The invention provides a haemoglobin of high yield active spore and a culture method thereof, belongs to the technical field of microorganisms, and particularly provides a haemoglobin strain HSJ1029 which is preserved in China general microbiological culture Collection center (China general microbiological culture Collection center) for 9 months and 14 days in 2023, and has the address of North Star Xiya No. 1,3 in the Beijing Kogyo area, and the preservation number of CGMCC No. 40866.

Inventors

  • YANG PENG
  • PENG LIZENG
  • HAN JIANDONG
  • PENG CHUNE
  • Lodi Ratna Silvia
  • HUANG CHUNYAN
  • LI YUNMEI

Assignees

  • 山东省农业科学院

Dates

Publication Date
20260508
Application Date
20260202
Priority Date
20250417

Claims (10)

  1. 1. The haemoglobin bacterium (Trametes sanguinea) HSJ1029 is preserved in China general microbiological culture Collection center (China general microbiological culture Collection center) with the address of XILU No. 1 and No. 3 of the Korean area North Star of Beijing city at the 9 th month 14 of 2023, and the preservation number is CGMCC No.40866.
  2. 2. The culture method of the high-yield haemoglobus spores is characterized by comprising the following steps of: Inoculating the haemoglobus hystericus HSJ1029 of claim 1 onto a modified PDA culture medium for culturing, and collecting spores after culturing until spores are produced; The improved PDA culture medium is prepared by adding maltose 0-30g/L and yeast extract 0-5g/L into PDA culture medium, and pH is 4-8.
  3. 3. The method according to claim 2, wherein the culture conditions are 28 to 30 ℃; Preferably, the culture conditions are 30 ℃.
  4. 4. The culture method according to claim 2, wherein the modified PDA culture medium is prepared by adding 20g/L maltose and 4g/L yeast extract to PDA culture medium, and has pH of 5.
  5. 5. The method of culturing of claim 2, wherein the method of spore collection comprises the steps of: Adding culture medium with spores into siloxane solution for treatment to disperse spores into siloxane solution, removing culture medium and mycelium to obtain liquid containing spores, centrifuging the liquid containing spores, and freeze-drying to obtain spores; Preferably, the spore-containing liquid is centrifuged at 8000-10000r/min for 8-10min.
  6. 6. The method of claim 5, wherein the concentration of the siloxane solution is 0.01% to 0.08% by volume.
  7. 7. The culture method as claimed in claim 5, wherein the spores are dispersed in a silicone solution by adding a medium having spores grown therein to a 0.01% silicone solution, then removing the medium and mycelia to obtain a liquid containing spores, then adding a 0.08% silicone solution to the medium and mycelia to treat the liquid containing spores, then removing the medium and mycelia again to obtain a liquid containing spores, and centrifuging all the obtained liquid containing spores at 10000r/min for 10min, and freeze-drying to obtain spores.
  8. 8. The method of claim 5, wherein the siloxane is a poly (dimethylsiloxane) hydride-terminated.
  9. 9. The culture method according to claim 2, wherein the haemoglobus HSJ1029 is inoculated in a cake form onto a modified PDA medium for culturing.
  10. 10. A composition comprising the spore powder of haemoglobus HSJ1029 of claim 1 and/or an extract of spore powder of haemoglobus HSJ 1029; preferably, the spore powder of the haemoglobus HSJ1029 is wall-broken spore powder; Preferably, the composition has at least one of immunomodulating, antibacterial or antioxidant effects.

Description

Haemophilus with high yield of active spores and culture method thereof Technical Field The invention belongs to the technical field of microorganisms, and particularly relates to a haemoglobin having high yield of active spores and a culture method thereof. Background The haemoglobus is a common medicinal fungus belonging to genus trametes of Polyporaceae of class Agaricales of basidiomycetes. The haemoglobin has wide distribution range in China, is mainly concentrated in Jilin, hebei, fujian and other places, and is mainly dried after picking in summer and autumn, and the fruiting body of the haemoglobin has high medicinal value, and can stop bleeding, detoxify, remove dampness, resist tumors and the like. Haemoglobins are also white rot fungi, sensitive to the ratio of carbon source to nitrogen source, and conventional media (such as potato dextrose agar medium, PDA for short) may not be effective in inducing sporogenesis. In the prior art, the flat plate culture of the haemoglobin has a plurality of other technical problems, such as long spore production period (for example, a strain of the haemoglobin Trametes sanguinea WTFA for producing laccase is disclosed in plasma mutagenesis Trametes sanguinea WTFA laccase production medium optimization and enzyme property research), the strain is inoculated on a YPD solid flat plate, 14d spore production is performed, spore collection is difficult, and spore activity and uniformity are poor. At present, the existing plate culture conditions of the haemoglobus are single, the screening effect is poor, and most researches on fruiting bodies and liquid fermentation broth are concentrated, so that spore activity and yield are limited, and research on the haemoglobus spores is hindered. The haemoglobus (Trametes sanguinea) has a related report, is mainly obtained by field collection, has extremely low proportion of fungus substances existing in the nature, has fewer developed types and is urgently required to be developed and utilized for a large amount of wild strain resources. The fungi are even the same genus and the same species, and the growth characteristics, metabolic activities and the like of the strains are obviously different. For example, "identification and domestication cultivation of a wild Polyporus strain" (Hebei society of science and technology, gao Zhiyuan, shuoshi paper) discloses that a wild Polyporus strain XHMK is trametes hemsleyanum Trametes sanguinea, the most suitable carbon source of the strain is fructose, the most suitable nitrogen source is yeast extract powder, the mycelium grows fastest at pH7.0, and the mycelium grows fastest at 35 ℃. A strain of haemoglobus (Trametes sanguinea) is disclosed in deep fermentation research of Wuling mountain haemoglobus (2010, 21 st, 6 th, changjiang society of life sciences and technology, chen Jinchao, wang Xinhui and the like), an optimal carbon source is glucose, an optimal nitrogen source is beef extract, the mycelium yield is highest at pH6.0, and the mycelium yield is highest at 25 ℃. In the prior art, the researches on laccase production and polysaccharide production of the haemoglobus are relatively more, but the researches on preparation of the haemoglobus spores are not reported, and the haemoglobus spores are used as main propagation organs of fungi because the haemoglobus is a common medicinal fungus and the fruiting body of the haemoglobus has high medicinal value, and the researches are also of great significance. Disclosure of Invention Aiming at the defects of the prior art, the invention provides the haemoglobin having high yield of active spores and a culture method thereof. The haemoglobin bacterium (Trametes sanguinea) HSJ1029 is preserved in China general microbiological culture Collection center (China general microbiological culture Collection center) with the address of XILU No.1 and No. 3 of the Korean area North Star of Beijing city at the 9 th month 14 of 2023, and the preservation number is CGMCC No.40866. A method for culturing high-yield hemoglobus spores comprises the following steps: Inoculating the haemoglobus HSJ1029 to an improved PDA culture medium for culturing until spores are generated, and collecting the spores; The improved PDA culture medium is prepared by adding maltose 0-30g/L and yeast extract 0-5g/L into PDA culture medium, and pH is 4-8. According to the invention, the culture conditions are preferably 28-30 ℃. Further preferably, the culture conditions are 30 ℃. According to a preferred embodiment of the present invention, the modified PDA culture medium is prepared by adding 20g/L maltose and 4g/L yeast extract to PDA culture medium, and the pH is 5. The PDA culture medium can be prepared according to a conventional formula or can be prepared by using a commercial PDA culture medium. According to a preferred embodiment of the invention, the method for collecting spores comprises the steps of: adding culture medium with spores into siloxane solution, tr