Penicillium purpurescens QL-9204 and application for preparing phloretin during phlorizin conversion

A QL-9204, Penicillium violaceum technology, applied in biochemical equipment and methods, microorganism-based methods, microorganisms, etc., can solve the problems of less conversion by-products, low conversion yield, poor specificity, etc. Easy, high conversion yield, batch stable effect

Active Publication Date: 2016-05-04
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a microbial strain producing glycosidase—Penicillium purpurogenum (Penicillium purpurogenum) QL-9204, and its application in converting phlorizin to prepare phloretin, which can overcome the existing acid-base hydrolysis method Compared with the problems of poor specificity and low conversion yield in the preparation of enzymatic hydrolysis, this process has the advantages of low cost, simple process, high conversion yield and less conversion by-products.

Method used

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  • Penicillium purpurescens QL-9204 and application for preparing phloretin during phlorizin conversion
  • Penicillium purpurescens QL-9204 and application for preparing phloretin during phlorizin conversion
  • Penicillium purpurescens QL-9204 and application for preparing phloretin during phlorizin conversion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Enrichment and isolation of transformed bacterial strains

[0024] Add about 50 g of crushed fresh apple peel into a 250 mL Erlenmeyer flask, and incubate at a constant temperature of 28°C for 5 days. Dilute the moldy enrichment 1 x 10 with sterile water 6 After doubling, spread it on the PDA plate medium, culture at 28°C for 3 days, pick mold colonies with different shapes and colors and transfer them to PDA slant medium, and place them at 28°C for 3 days to obtain slant strains rich in spores The 11 strains were numbered (QL-9201~QL-9211) respectively, and stored in a refrigerator at 4°C for later use.

[0025] Both the plate medium and the slant medium are potato dextrose agar medium (PDA), prepared according to the following composition and method: potatoes are washed, peeled and cut into small pieces, weighed 200g, added tap water 1000mL, boiled for 30min, 4 Filter with a layer of gauze to remove slag, make up the filtrate to 1000mL, then add 20g of su...

Embodiment 2

[0026] Example 2: Screening and classification identification of transformed strains

[0027] Use an inoculation loop to pick 2 rings of spores of the slant strains of each strain obtained in Example 1, insert them into a 100mL fermentation medium (packed in a 250mL Erlenmeyer bottle), and cultivate them for 5 days at 28°C and 200r / min constant temperature shaking conditions for enzyme production , filtered with a Buchner funnel, and the collected filtrate is the crude enzyme solution. Take 50mL of crude enzyme solution and place it in a 250mL Erlenmeyer flask, dissolve 1mg of phlorizin in 0.1mL of methanol, and then add it to the crude enzyme solution to form a reaction system (total volume is 50mL), shake at 28°C and 200r / min After down-conversion for 8 hours; take 10 mL of the conversion solution and extract twice with the same volume of ethyl acetate, combine the ethyl acetate in a round-bottomed flask, evaporate the ethyl acetate to dryness under reduced pressure, dissolv...

Embodiment 3

[0034] Embodiment 3: the application of Penicillium purpurea QL-9204 strain transforming phlorizin to generate phloretin 1

[0035] The Penicillium purpurea QL-9204 screened and obtained in Example 2 was used as the transformed bacterial species, and the crude enzyme liquid prepared by fermentation was transformed into phloridzin through seed expansion cultivation, and the molar conversion yield of phloridin was slightly higher than that in Example 2. Improve, the result of repeated experiment 3 batches has no significant difference, shows that the performance that this bacterial classification transforms phloridin to produce phloretin is stable, and concrete process step is as follows:

[0036] (1) Inoculate the slant strain of Penicillium violaceum QL-9204 preserved in a refrigerator at 4°C in fresh PDA slant medium, and culture the slant at a constant temperature of 28°C for 3 days. Described PDA slant culture medium composition and preparation method are with embodiment 1;...

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Abstract

The invention discloses penicillium purpurescens QL-9204 and application for preparing phloretin during phlorizin conversion. The penicillium purpurescens QL-9204 is a strain free of toxicity and harm and safe to use. The penicillium purpurescens QL-9204 grows rapidly, and is high in hybrid bacteria resistance, easy to culture and stable in batch; a fermentation medium is simple in components and low in market price, thereby being low in production cost; a substrate plorizin is directly put into a crude enzyme for conversion, the enzyme separation and purification steps are omitted, the technology is simple, and the penicillium purpurescens QL-9204 is easy to apply industrially; the phloretin conversion yield is high and can reach 92.5% to the maximum, and the penicillium purpurescens QL-9204 has the advantages that few by-products are produced, and the product is easy to extract.

Description

(1) Technical field [0001] The invention relates to a method for preparing phloretin, in particular to a strain of Penicillium purpurogenum QL-9204, and the application of the strain in the field of transforming phloridzin to prepare phloretin. (2) Background technology [0002] Phloretin (phloretin), also known as trihydric phenol acetone, CAS No. 60-82-2, belongs to dihydrochalcone compounds. The chemical structure is 3-(4-hydroxyphenyl)-1-(2,4,6-trihydroxyphenyl)-1-propanone, and the molecular formula is C 15 h 14 o 5 , the molecular weight is 274.27, the pure product is pink, soluble in methanol, ethanol and acetone, almost insoluble in water, soluble in alkaline solution. Phloretin has good anti-oxidation and anti-free radical effects, can inhibit the activity of melanocytes, and has a good effect on lightening spots; not only that, but also has good anti-inflammatory and immunosuppressive effects; the latest Studies have shown that phloretin also has anti-tumor and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P7/26C12R1/80
CPCC12N1/14C12P7/26C12N1/145C12R2001/80
Inventor 梅建凤李靓金航应国清王鸿易喻
Owner ZHEJIANG UNIV OF TECH
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