Specific strain and application thereof

A technology of Pseudomonas chloropinus and Aspergillus niger, applied in the field of microorganisms, can solve the problems of high industrialization cost, weak conversion production capacity, and low production efficiency

Active Publication Date: 2019-03-29
BOTON SHANGHAI BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The known prior art has disclosed the ability of Aspergillus niger MIC 373 and Aspergillus niger CGMCC 0774 to transform ferulic acid to produce vanillic acid, but the existing known Aspergillus niger strains have weak transformation and production capacity, and the yield of vanillic acid is lower than 5g / L, making its production efficiency low and the industrialization cost too high

Method used

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  • Specific strain and application thereof
  • Specific strain and application thereof
  • Specific strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Embodiment 1 Aspergillus niger CFFSH005 strain identification

[0071] Aspergillus niger strain CFFSH005 was isolated from the root soil of watermelon in Nanhui, Pudong New District, Shanghai.

[0072] Aspergillus niger CFFSH007 grows on potato medium, the mycelia are colorless at the initial stage, and dark brown to nearly black when mature. Observing under a microscope after taking the mycelia for staining, the mycelia are separated and have black spherical sporangia. According to K.B.Raper and D.I.Fennell "Aspergillus" (The Genus Aspergillus, 1965), the strain has typical characteristics of Aspergillus niger. Aspergillus niger CFFSH005 was deposited in the China Center for Type Culture Collection on December 20, 2016, and the preservation number is CCTCC NO:M 2016767

Embodiment 2

[0073] Example 2 Identification of Pseudomonas chlororaphis CFFSH007 and Pseudomonas chlorophis CFFSH008 strains

[0074] Two strains of Pseudomonas chloropinus were isolated from plant roots, identified by 16s rDNA, and the general primers were synthesized by Suzhou Jinweizhi Biotechnology Co., Ltd. The 27F / 1492R sequence is as follows:

[0075] 27F (5'→3'): AGAGTTTGATCMTGGCTCAG

[0076] 1492R (5'→3'): TACGGYTACCTTGTTACGACTT

[0077] The colony of Pseudomonas chlororaphis CFFSH007 was taken, and its 16s rDNA was subjected to colony PCR with universal primers 27F and 1492R to obtain a full-length 1413bp DNA sequence, which is shown in sequence 1 of the gene sequence attached table. Using NCBI's Sequence 1 was compared, and the results showed that the similarity between Pseudomonas chlororaphis CFFSH007 and the 16s rDNA of Pseudomonas chlororaphis ATCC 13985 was 100% (1413 / 1413); at the same time, the similarity with the 16s rDNA of Pseudomonas chlororaphis Lzh-T5 was 100% (...

Embodiment 3

[0079] Example 3 Screening of strains producing vanillic acid

[0080] (1) Isolation of strains

[0081] Take 50g fresh root soil samples of watermelon field in Nanhui area, Shanghai, disperse and mix with 100mL sterile normal saline, take 100uL for gradient dilution, spread on the prepared sieving agar plate, and place in a 30°C incubator Incubate until a single colony is formed. The formula of the sieving agar plate culture medium is: 2% (w / v) agar powder, 5% (w / v) glucose, 0.5% (w / v) ferulic acid, 1% yeast powder, 0.3% pancreatic Peptone, pH 7.0. The formed single colonies were transferred and preserved, and the strains were identified.

[0082] (2) Identification of vanillic acid transformation ability of wild Aspergillus niger

[0083] Select to obtain 10 strains of Aspergillus niger strains from soil samples, get the mature spores cultivated in the potato dextrose solid medium plate for 7 days, and inoculate them in 90mL culture potato dextrose liquid medium (potato ...

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Abstract

The invention relates to the technical field of biology, and particularly relates to a specific strain and application thereof. Pseudomonas chlororaphis can generate vanillic acid by using ferulic acid as substrate through biotransformation. Aspergillus niger having a preservation number of CCTCC NO:M 2016767 is obtained by screening, has the content of vanillic acid being more than 10g / L. The strain with higher specificity can directly utilize vegetation raw materials, and the concentration of vanillic acid can be 0.43g / L.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to specific bacterial strains and applications thereof. Background technique [0002] 4-Hydroxy-3 methoxybenzoic acid (also known as vanillic acid, vanillic acid) is a class of important natural compounds. It is the main functional component of Chinese herbal medicine Huhuanglian and Angelica, and it is also widely found in gardenia, vanilla, etc. In natural plant extracts. Vanillic acid has natural antioxidant biological activity, and has broad application prospects in the fields of medical treatment, health care, and cosmetics. In the field of antioxidant drugs, vanillic acid has a good effect on reducing and preventing cell damage. For example, in the classic cancer chemotherapy drug cisplatin chemotherapy regimen, the combined application of vanillic acid can greatly reduce the nephrotoxicity of the chemotherapy drug cisplatin (CDDP) and its damage to the antioxidant de...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P39/00C12P7/42C12N1/15C12N1/21C12R1/38C12R1/685
CPCC12N9/0006C12P7/42C12P39/00
Inventor 张鹏何汉平
Owner BOTON SHANGHAI BIOLOGICAL TECH CO LTD
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