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Pseudomonas capable of producing pyrroloquinoline quinone and application thereof

A pyrroloquinoline quinone and Pseudomonas technology, applied in the direction of bacteria, microbe-based methods, microbes, etc., can solve the problem of low PQQ production, and achieve the effect of avoiding flammability and explosion and rapid growth

Active Publication Date: 2021-09-03
BLOOMAGE BIOTECHNOLOGY CORP LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Master's thesis "Study on the Synthesis of Pyrroloquinoline Quinone by Gluconobacter oxidans" used 50g / L glucose as carbon source, and used Gluconobacter oxidans for fermentation, but its PQQ yield was low, the highest being 0.813mg / L

Method used

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  • Pseudomonas capable of producing pyrroloquinoline quinone and application thereof
  • Pseudomonas capable of producing pyrroloquinoline quinone and application thereof
  • Pseudomonas capable of producing pyrroloquinoline quinone and application thereof

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specific Embodiment approach

[0047] As a specific embodiment of the present invention, a composition is obtained by fermenting Pseudomonas HQ-2 of the present invention, which comprises the above-mentioned Pseudomonas HQ-2 and fermentation broth, so The concentration of pyrroloquinoline quinone in the fermentation broth is as high as 5.00 mg / L or more, specifically 10.00 mg / L or more, 15.00 mg / L or more, 20.00 mg / L or more, 25.00 mg / L or more, 30.00 mg / L or more , more than 35.00 mg / L, more than 40.00 mg / L, more than 45.00 mg / L, more than 50.00 mg / L, more than 55.00 mg / L or more than 60.00 mg / L, preferably more than 60.00 mg / L.

[0048] In a specific embodiment, the fermentation medium used to produce the fermentation broth comprises a carbon source, a nitrogen source, and an inorganic salt, wherein the carbon source can be selected from one or a combination of glucose and sucrose, and the nitrogen source can be One or a combination of two or more selected from peptone, yeast powder, soybean peptide, ammo...

Embodiment 1P

[0073] The screening of embodiment 1PQQ producing bacteria

[0074] A total of 61 soil or water samples were collected from Shandong, Henan, and Inner Mongolia. After being enriched by the screening medium, they were diluted and spread on the screening medium plate, and cultured at 30°C for 3 to 5 days to obtain the primary screening strain. The primary screening strains were inoculated into test tubes containing 5mL of screening medium, cultured with shaking at 30°C for 2 days, then transferred to a Erlenmeyer flask containing 50mL of screening medium, and fermented at 30°C and 200rpm for 3 to 5 days. As shown in Table 1, the content of PQQ in the fermentation supernatant was accurately determined by HPLC, and the strain numbered Pseudomonas.spHQ-2 had the highest yield, reaching 22.32 mg / L.

[0075] Table 1 Different isolates producing PQQ

[0076] Strain number PQQ production (mg / L) Strain number PQQ production (mg / L) HQ-1 35.19 HQ-8 18.32 HQ-...

Embodiment 2

[0077] Embodiment 2 morphology observation

[0078] For the morphological characteristics of Pseudomonas (Pseudomonas.sp) HQ-2, see the attached figure 1 . Specifically, it was observed with an OLYMPUSCX33 microscope, and the bacterial cells were observed to be in the shape of short rods.

[0079] The colony morphological characteristics of Pseudomonas (Pseudomonas.sp) HQ-2 are detailed in the attached figure 2 . It can be observed that the colonies are off-white, flat, moist, and round.

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Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to pseudomonas and a method for producing pyrroloquinoline quinine through fermentation of the pseudomonas. According to the invention, a new strain Pseudomonas.sp is screened from soil near a pharmaceutical factory, the new strain is named as HQ-2, and the preservation number of the strain is CCTCC NO: M 2021251. The invention also relates to an application of the pseudomonas (Pseudomonas sp) HQ-2 in fermentation production of the pyrroloquinoline quinone, a method for preparing the pyrroloquinoline quinone, and a composition thereof. The strain is aerobically cultured for 1-2 days in a culture medium taking cane sugar and glucose as carbon sources, and the yield of pyrroloquinoline quinone can reach 60mg / L at a shake flask level. The strain screened by the invention can secrete PQQ out of cells, and compared with a common PQQ producing strain, the strain has the characteristics of rapid growth and capability of utilizing sucrose and glucose, and a new strain is provided for industrial fermentation production of PQQ.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a pyrroloquinoline quinone-producing pseudomonas and application thereof. Background technique [0002] Pyrroloquinoline quinone (Pyrroloquinoline quinone, PQQ, also known as Methoxatin) is a water-soluble quinone compound with heat stability. It was first identified as a cofactor of alcohol and glucose dehydrogenase in bacteria and is a flavin nucleotide After nicotinamide nucleotide, the third prosthetic group found in bacterial dehydrogenase, the world medical community calls it the fourteenth vitamin. As a new type of water-soluble vitamin, PQQ is a prosthetic group of oxidoreductase, which is very rare and exists in some microorganisms, plants and animal tissues. Experimental mice lacking this substance will have low reproductive ability and other phenomena. Studies have shown that PQQ exists in daily vegetables and fruits, such as natto, celery and kiwi. This subs...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P17/18C12R1/38
CPCC12N1/20C12P17/182
Inventor 乔莉苹周宁李帆王秀娟沈延臻郭学平
Owner BLOOMAGE BIOTECHNOLOGY CORP LTD
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