Pyrroloquinoline-quinone-producing Hyphomicrobium strain and application thereof

A technology of pyrroloquinoline quinone and filamentous microbe, which is applied in the directions of bacteria, microorganisms, microorganisms, etc., can solve problems such as yield decline, and achieve the effect of stable fermentation performance and favorable purification.

Active Publication Date: 2015-07-01
福建力多利生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, as a coenzyme, PQQ can irreversibly react with amino acids in the culture medium to form stable inactive oxazole compounds, which will cause a sharp drop in production

Method used

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  • Pyrroloquinoline-quinone-producing Hyphomicrobium strain and application thereof
  • Pyrroloquinoline-quinone-producing Hyphomicrobium strain and application thereof
  • Pyrroloquinoline-quinone-producing Hyphomicrobium strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] The composition of the fermentation medium in embodiment 1: methanol 20g / L, ammonium sulfate 2g / L, potassium dihydrogen phosphate 1.2g / L, disodium hydrogen phosphate 4g / L, magnesium sulfate 1g / L, trace element solution 1mL / L L. Vitamin auxiliary solution 2mL / L, the balance is distilled water, pH 6.8.

[0063] Embodiment 1, the screening of pyrroloquinoline quinone producing bacteria

[0064] 1. The chemical sewage water samples collected from Quangang Petrochemical Zone, Quanzhou City were cultured with enrichment medium at 30°C for 48 hours to obtain a culture solution, which was diluted and spread on a screening medium plate, and a single colony (methyl After vegetative bacteria), single colonies were picked and inoculated in the fermentation medium, 30°C, 220r / min shaking culture for 96h, and the growth of the bacteria was observed. Among the more than 120 isolates of methylotrophs, 47 strains could change the color of the screening medium (red, brown or yellow).

...

Embodiment 2

[0079] Embodiment 2, identification of bacterial strain FJNU-R8

[0080] 1. According to "Bergey's Handbook of Systematic Bacteriology" (Second Edition, 2004), the numbered FJNU-R8 strain obtained by screening in Example 1 was identified for its morphological characteristics and physiological and biochemical characteristics. The results are shown in Table 2.

[0081] FJNU-R8 strain is aerobic, Gram-negative, suitable growth temperature is 25-32°C, suitable growth pH is 6.0-8.0; can utilize arabinose, sucrose, xylose, fructose, trehalose and raffinose, but cannot Alcohols such as mannitol, arabitol, and inositol, and cannot utilize amino acids such as ornithine, arginine, and lysine; positive for oxidase, negative for urease and tryptophan deaminase; methyl red, V-P, and starch The hydrolysis test is negative; it can reduce nitrite without producing indole and hydrogen sulfide; it has no fluorescence reaction.

[0082] 2. The colonies of the FJNU-R8 strain are small, round, regu...

Embodiment 3

[0090] Embodiment 3, the influence of culture time on pyrroloquinoline quinone output

[0091] One, pick the FJNU-R8 single bacterium colony of embodiment 2 in the Erlenmeyer flask that 30mL of the seed medium shown in A below is housed, 30 ℃, 220rpm shaking culture 24-36 hour (OD 650 Between 2.5-3.0, adjust to zero with the seed medium) to obtain the seed solution; then transfer the inoculum of 5% of the seed solution by volume to a 250mL Erlenmeyer flask equipped with 50mL of the fermentation medium shown in B below in 30°C, 150-220rpm (Day 0-2: 220rpm; Day 2-6: 150rpm) for 5-6 days to obtain a fermentation broth, which was taken every 8 hours from the start of the transfer, Determination of the biomass of the fermentation broth (OD 650 , adjusted to zero with the fermentation medium shown in B), and the fermentation broth was centrifuged at 10000 × g for 10 min, and the supernatant was taken to measure the pyrroloquinoline quinone (PQQ) yield, methanol and protein content ...

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Abstract

The invention discloses a pyrroloquinoline-quinone-producing Hyphomicrobium strain and application thereof. The invention discloses a Hyphomicrobium denitrificans strain of which the collection number is CGMCC No.10620. The culture solution of the pyrroloquinoline-quinone-producing Hyphomicrobium FJNU-R8 has higher pyrroloquinoline quinone yield. After the strain is cultured in a culture medium using methanol as a carbon source for 4-6 days, the pyrroloquinoline quinone yield is 150-350 mg / L. The protein content is low, and the fermenting property is stable. The Hyphomicrobium disclosed by the invention can be used in producing pyrroloquinoline quinone by microbial fermentation.

Description

technical field [0001] The invention relates to a pyrroloquinoline quinone-producing mycelium and application thereof, belonging to the field of biotechnology. Background technique [0002] Pyrroloquinoline quinone (PQQ) is a reddish-brown water-soluble ortho-quinone compound that is easily decomposed by exposure to light and is thermally stable. It is the third oxidoreductase discovered after NAD / NADP and FMN / FAD (oxidordeuctase) coenzyme, involved in respiratory chain electron transfer. The earliest research on PQQ can be traced back to 1959 when Hauge found that the glucose dehydrogenase activity of Acinetobacter calcoaceticus did not depend on pyridine nucleotides and flavin nucleotides, but on a new separable coenzyme. In 1979, Duine et al. used electron spin resonance, nuclear magnetic resonance and mass spectrometry to find that this coenzyme was an ortho-quinone compound. Subsequently, Salisbury (1979) clarified the chemical structure of the coenzyme by using X-ra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P17/18C12R1/01
CPCC12P17/182C12N1/205C12R2001/01
Inventor 柯崇榕黄建忠杨诗颖杨欣伟钟璐芳
Owner 福建力多利生物科技有限公司
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