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Methylovorus.sp capable of producing pyrroloquinoline quinone and application thereof

A technology of pyrroloquinoline quinone and methyl-eating bacteria, which can be applied to bacteria, microorganism-based methods, microorganisms, etc., and can solve problems such as difficulty in industrial-scale production

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

[0006] Patent CN102061278B discloses the application of Methylovorus sp. MP688 to produce pyrroloquinoline quinone (PQQ), and its PQQ output reaches 125mg / L, but the culture time of the strain is 5 days, which is relatively long, and realizes industrial scale production There is a certain degree of difficulty

Method used

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  • Methylovorus.sp capable of producing pyrroloquinoline quinone and application thereof
  • Methylovorus.sp capable of producing pyrroloquinoline quinone and application thereof
  • Methylovorus.sp capable of producing pyrroloquinoline quinone and application thereof

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Embodiment approach

[0037] As a specific embodiment of the present invention, it relates to a Methylovorus.sp HQ-1, which has been preserved in the China Center for Type Culture Collection (CCTCC) on March 19, 2021, and the preservation address is Wuhan, China, Wuhan University, postcode: 430072, deposit number CCTCCNO: M 2021252, which can be used for fermentative production of pyrroloquinoline quinone.

[0038] Pyrroloquinoline quinone, the English name is methoxatin, referred to as PQQ, the molecular formula is C 14 h 6 N 2 o 8 , with a molecular weight of 330.206 and a CAS registration number of 72909-34-3, a chemical intermediate. PQQ is a new redox prosthetic group, which has various physiological functions such as anti-oxidation, regulating the level of free radicals in the body, and promoting the regeneration of nerve growth factors.

[0039] The 16s rRNA gene sequence of the methylotroph HQ-1 is SEQ ID NO: 1, see attached Figure 4 .

[0040] rRNA has the characteristics of uniquen...

Embodiment 1P

[0067] The screening of embodiment 1PQQ producing bacteria

[0068] 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 strains. The primary screening strains were inoculated into test tubes containing 5 mL of screening medium, cultured with shaking at 30°C for 2 days, then transferred to a Erlenmeyer flask containing 50 mL of screening medium, and cultured at 30°C and 200 rpm 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 Methylovorus.sp HQ-1 had the highest yield, reaching 35.19 mg / L.

[0069] Table 1 Different isolates producing PQQ

[0070] Strain number PQQ production (mg / L) Strain number PQQ production (mg / L) HQ-1 35.19 HQ-8 18.32 ...

Embodiment 2

[0071] Embodiment 2 morphology observation

[0072] For the morphological characteristics of Methylovorus.sp HQ-1, see the attached figure 1 . Specifically, it was observed with an OLYMPUS CX33 microscope, and the bacterial cells were observed to be in the shape of short rods.

[0073] The colony morphological characteristics of Methylovorus.sp HQ-1 are detailed in the attached figure 2 . It can be observed that the colonies are large, flat, moist, round, and pale yellow in color.

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Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to a methylovous.sp HQ-1 with the preservation number of CCTCC (China Center For Type Culture Collection) NO: M2021252. The invention also relates to an application of the Methylovous.sp HQ-1 in fermentation production of the pyrroloquinoline quinone, a method for preparing the pyrroloquinoline quinone, and a composition thereof. The methylovous.sp HQ-1 provided by the invention is aerobically cultured for 1-2 days in a culture medium taking methanol as a carbon source and peptone as a nitrogen source, and the yield of pyrroloquinoline quinone can reach 100mg / 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 advantage of rapid growth, 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 methyl-eating fungus producing pyrroloquinoline quinone and an 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...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P17/18C12R1/01
CPCC12P17/182C12N1/20Y02E50/30
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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