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Fungal production of fdca

A fungal cell and wild-type technology, applied in the fields of molecular genetics, biotransformation and fermentation, and metabolic engineering, can solve the problems of FDCA produced by yeast or filamentous fungi that have not been described

Inactive Publication Date: 2018-08-03
PURAC BIOCHEM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the production of FDCA by yeast or filamentous fungi has not been described

Method used

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  • Fungal production of fdca
  • Fungal production of fdca
  • Fungal production of fdca

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0381] Example 1: Isolation of Penicillium brasiliensis Batista

[0382] Fungal strains with the ability to grow at low pH to consume HMF were enriched and isolated from Dutch soil. 1 g of air-dried soil was provided to contain 20 ml of K-omitted 2 HPO 4 solution in a 100ml Erlenmeyer flask. The pH of the medium was lowered to pH=3 by titration with HCl solution. The initial HMF concentration was 1 g / l. To inhibit bacterial growth, naladixic acid (20 mg / l) was included in the medium. The vials were incubated at 30°C under air. After 2 weeks of incubation, the material (1 ml) from the enriched culture was transferred to fresh medium with the same initial composition. From the second culture, agar plates were streaked, containing mineral medium and 0.5 g / l HMF. Colonies that emerged after 10 days of incubation at 30°C under air were restreaked on plates with the same initial medium until the organisms were purified. Isolates were tested for HMF-depleted growth by growi...

Embodiment 2

[0385] Example 2: Limitation of Penicillium brasiliensis Growth at Low pH

[0386] The Penicillium brasiliensis C1 strain was isolated by incubating the soil in an initial pH=3 medium. However, pH changes in the medium during growth also varied in subsequent batch experiments with pure cultures. To verify the pH limitation of growth, chemostat cultures were performed to allow a predetermined growth rate at a constant predetermined pH. The growth rate was set at 0.08h -1, and the pH was maintained at pH=2.9. In 3 separate experiments, HMF, HMFCA or FDCA were used as carbon source. Under each of the three conditions the organism was able to establish itself at steady state; no washout occurred. The concentration of any of the 3 furans in the culture vessel was below the limit of detection in any of the three runs. The results show the ability of the organism to grow at pH values ​​below 3, which is a desirable property for FDCA production from HMF at low pH values. In ad...

Embodiment 3

[0387] Example 3: Identification of the Penicillium brasiliensis C1 gene encoding an enzyme involved in HMF catabolism

[0388] Sequencing and Annotation of the Penicillium brasiliensis C1 Genome

[0389] DNA from Penicillium brasiliensis C1 was isolated and sent to BaseClear for paired-end sequencing using the Illumina HiSeq2500 system. After quality checking, filtering, and adapter removal, read sequences were assembled into contigs and ligated and placed in scaffolds.

[0390] Genome annotation was performed using the BaseClear annotation pipeline based on Augustus for structural annotation (Stanke and Waack, 2003, Bioinformatics.19 Supp1 2:ii215-25) and the Prokka Prokaryotic Annotation system for functional annotation (http: / / vicbioinformatics .com) combination. A set of Penicillium species was used as a reference for annotations that included information on rRNA, tRNA, signal peptides, Pfam protein family predictions, cellular localization, and conserved domains.

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Abstract

The invention relates to fungal cells for the production of FDCA. The fungal cell is genetically modified to have at least one of a) a genetic modification that confers to or increases in the cell theability to oxidize 5-hydroxymethyl-2-furancarboxylic acid to 5-formyl-2-furoic acid; and, b) a genetic modification that reduces catabolism of 2,5-furandicarboxylic acid in the cell. The fungal cellcan further be genetically modified to increase the cell's ability to oxidize furanic aldehydes to the corresponding furanic carboxylic acids. The invention also relates to a process for the production of 2,5-furandicarboxylic acid (FDCA) wherein the cells of the invention are used for oxidation of a furanic precursors of FDCA.

Description

technical field [0001] The invention relates to the technical fields of molecular genetics, metabolic engineering, biotransformation and fermentation. In particular, the present invention relates to the production of 2,5-furandicarboxylic acid from hydroxymethylfurfural by genetically modified fungi. The invention further relates to the use of this fungus in a process for the bioconversion of hydroxymethylfurfural to 2,5-furandicarboxylic acid. Background technique [0002] 2,5-Furandicarboxylic acid (FDCA) is a monomeric compound that can be used to create polyesters with enormous economic impact. A very important compound in this field is polyethylene terephthalate (PET), produced from terephthalic acid (PTA) and ethylene glycol. FDCA can replace PTA in polyester PET, in this case yielding polyethylene furandicarboxylate (PEF). PEF has good potential to replace PET in the large polyester market. Not only because of its superior properties when compared to PET, but also...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/06C12P1/02C12P7/44
CPCC12N9/0006C12N9/0073C12P7/44C12N15/52C12P7/42Y02E50/10C12P1/02
Inventor J·A·M·德邦塔H·J·鲁伊塞纳尔斯J·韦赖伊
Owner PURAC BIOCHEM
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