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Active extract of facultative anaerobic sea Pseudomonas stuszeri as well as production method and use thereof

A Pseudomonas stutzeri, facultative anaerobic technology, applied in the field of active extracts of facultative anaerobic marine denitrifying bacterial cultures

Inactive Publication Date: 2009-08-05
DALIAN JIAOTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The denitrifying bacteria in seawater belong to the group of facultative anaerobic bacteria. These microorganisms perform aerobic respiration under aerobic culture conditions and nitrate respiration under anoxic culture conditions. Many secondary metabolites are different from aerobic respiration, but so far there is no report to isolate and extract pharmacologically active components from marine denitrifying bacterial taxa in anoxic culture

Method used

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  • Active extract of facultative anaerobic sea Pseudomonas stuszeri as well as production method and use thereof
  • Active extract of facultative anaerobic sea Pseudomonas stuszeri as well as production method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1. liquid culture medium is:

[0049] Sodium acetate 20.0g;

[0050] KNO 3 2.0g;

[0051] K 2 HPO 4 0.5g;

[0052] MgSO 4 0.2g;

[0053] NaCl 25.0g;

[0054] MgSO 4 ·7H 2 O 3.0g;

[0055] CaCl 2 2H 2 O 0.05g;

[0056] Distilled water 1000ml;

[0057] Adjust the pH to 7.2±0.2 with sodium hydroxide at a concentration of 0.1 mol / L.

[0058] A. Seed culture: Fill the liquid medium into the serum bottle, leave a gap of 2cm in the bottle, tighten the cap of the serum bottle slightly to facilitate ventilation, and sterilize in a high-pressure sterilizer at 121°C and 0.105Mpa for 20min; after cooling Inoculate the marine strain DN7, fill it up with sterile medium, and tighten the bottle cap tightly; place the serum bottle in a constant temperature incubator for cultivation at 35-37°C. The culture time is 10-15 days;

[0059] B. Expanded cultivation: inoculate with 5-10% seed culture solution...

Embodiment 2

[0062] Embodiment 2. liquid culture medium is:

[0063] Sodium Potassium Tartrate 20.0g

[0064] KNO 3 2.0g

[0065] K 2 HPO 4 0.5g

[0066] MgSO 4 0.2g

[0067] Artificial sea water (or old sea water) 1000ml

[0068] Use sodium hydroxide with a concentration of 0.3mol / L to adjust the pH to 7.2-7.5.

[0069] Artificial Seawater Formula:

[0070] NaCl 25.0g

[0071] Na 2 SO 4 4.0g

[0072] KCl0.7g

[0073] NaHCO 3 0.20g

[0074] KBr 0.10g

[0075] h 3 BO 3 0.03g

[0076] NaF0.003g

[0077] 1.0mol / L MgCl 2 Solution 53mL

[0078] 1.0mol / l CaCl 2 Solution 10mL

[0079] 0.1mol / L SrO 2 Solution 0.90ml

[0080] Distilled water 1000ml

[0081] Use sodium hydroxide with a concentration of 0.2mol / L to adjust the pH to 7.2-7.5.

[0082] A. Seed culture: Fill the liquid medium into the serum bottle, leave a gap of 2cm in the bottle, tighte...

Embodiment 3

[0086] Example 3. Active component activity assay results

[0087] Using the agar dilution method (National Committee for Clinical Laboratory Standards. 2000. Approved standard: M2 ~ A7, 7th ed. NCCLS, Wayne, Pa.) to determine the MIC value of Pseudomonas aeruginosa and Bacillus subtilis, confirm the culture medium The ethyl acetate extract phase has antibacterial activity: the MIC values ​​of the prepared active components against Pseudomonas aeruginosa were 0.64 μg / mL, and the MIC values ​​against Bacillus subtilis were 1.28 μg / mL.

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Abstract

The invention relates to an active extract for facultative anaerobic marine Pseudomonas stutzeri and a preparation method and application thereof. A marine strain DN7 which is separated from the Bohai Sea of China is identified as the Pseudomonas stutzeri. An n-butyl alcohol extract phase of a culture solution after the propagation of the DN7 under the anoxic condition is subjected to silica gel thin-layer chromatography and reversed high-efficiency liquid phase chromatographic resolution to obtain white powder of two effective parts, and the white powder of the two effective parts has antibacterial activity on pseudomonas aeruginosa and Bacillus subtilis and is potentially used as an antibacterial medicine.

Description

technical field [0001] The invention relates to a pharmaceutical preparation derived from bacteria, in particular to an active extract derived from a culture of facultative anaerobic marine denitrifying bacteria and a preparation method thereof. Background technique [0002] The development of new drugs derived from marine microorganisms is a worldwide research hotspot in the 21st century. It has attracted widespread attention due to its characteristics of new strain sources, production of new structural active substances, overcoming traditional antibiotic resistance, and realizing resource sustainability. . At present, many novel antibacterial antibiotics, antifungal antibiotics, antiviral antibiotics, antitumor antibiotics, and some anti-inflammatory and analgesic active substances, enzyme inhibitors, etc. have been isolated from various marine microorganisms. However, at present, the research and development objects of new life active substances are mainly focused on mar...

Claims

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Application Information

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IPC IPC(8): A61K35/74A61P31/04C12P1/04
Inventor 穆军顾晓洁张翼付步飞董学伟
Owner DALIAN JIAOTONG UNIVERSITY
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