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Aspergillus niger JH-2 and application to biotransformation and synthesis of asiatic acid

A JH-2, Aspergillus niger technology, applied in biochemical equipment and methods, microorganisms, microorganisms, etc., can solve problems such as increasing process complexity, and achieve the effects of eliminating separation and purification steps, high conversion rate, and low production cost.

Active Publication Date: 2015-02-18
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, it has been reported that the crude extract of asiaticoside is converted by microorganisms, and a good conversion rate can be obtained, but the reported methods all require crude extraction of asiaticoside, which increases the complexity of the process

Method used

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  • Aspergillus niger JH-2 and application to biotransformation and synthesis of asiatic acid
  • Aspergillus niger JH-2 and application to biotransformation and synthesis of asiatic acid
  • Aspergillus niger JH-2 and application to biotransformation and synthesis of asiatic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Enrichment and isolation of transformed microbial strains

[0029] Add about 10g of Centella asiatica powder (commercially available), a traditional Chinese medicinal material that has not been sterilized by dry heat, into a 250mL conical flask, add 10mL of sterile water, stir evenly, and place it in an incubator at 28°C for 5 days to convert asiaticoside Enrichment culture of microbial strains until the centella asiatica powder is covered with mold.

[0030] Dilute the enriched culture full of mold with sterile water and spread it on the plate medium, culture it in a biochemical incubator at 28°C for 2 days, pick mold colonies with different shapes and colors and transfer them to the slant medium, Cultured at a constant temperature of 28°C for 3 days, 12 spore-rich slant strains were obtained, numbered JH-1~JH-12, and stored in a refrigerator at 4°C for later use.

[0031] Both the plate medium and the slant medium are potato dextrose agar medium (PDA), pre...

Embodiment 2

[0032] Embodiment 2: Screening of transformed microbial strains

[0033] Centella asiatica powder pretreatment: Centella asiatica powder was crushed through a 75-mesh sieve, and then sterilized by dry heat at 160°C for 2 hours.

[0034] Pick 2 rings of spores of each bacterial strain slant bacterial classification obtained in Example 1 respectively with an inoculation loop, insert in 20mL transformation medium (50mL Erlenmeyer bottle), add 1g of pretreated Centella asiatica powder, at 30 ℃, After 6 days of fermentation and cultivation under the shaking condition of 200r / min, filter with a Buchner funnel, and dry the centella asiatica powder and bacteria on the filter paper at 85°C for 24 hours. After drying, put all the dry matter into a 50mL Erlenmeyer flask. Add 20 mL of 95% ethanol aqueous solution, stand still for extraction for 1 h, and then extract for 1 h at 25° C. and 100 KHz ultrasonic conditions. After the extraction is completed, filter with suction to obtain an et...

Embodiment 3

[0040] Embodiment 3: the application of Aspergillus niger JH-2 strain in improving the content of asiatic acid in Centella asiatica 1

[0041] Using the Aspergillus niger JH-2 obtained by screening in Example 2 as the transformed strain, through seed expansion cultivation, and transforming Centella asiatica, the range of increase in the content of Asiatic acid is equivalent to that in Example 2, indicating that the strain transforms Asiaticoside The performance is stable, and the specific process steps are as follows:

[0042] (1) Inoculate the slant strain of Aspergillus niger JH-2 preserved in the refrigerator at 4°C in a fresh slant medium, and culture the slant in a biochemical incubator at 28°C for 3 days. The composition and preparation method of the slant culture medium are the same as in Example 1.

[0043] (2) Use an inoculation loop to pick up two rings of Aspergillus niger JH-2 spores after activation in step (1) into the seed medium, and culture them at 30°C and 2...

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Abstract

The invention provides a glycosidase producing strain, that is, Aspergillus niger JH-2 and an application of the Aspergillus niger JH-2 to biotransformation and increasing of the content of asiatic acid in centella asiatica. The Aspergillus niger is a common strain in the fermentation industry and is non-toxic, harmless and safe to use, the converting culture medium is simple in component, only five salt components are needed, and the production cost is low; the Aspergillus niger JH-2 grows fast and is high in infectious microbe resistance, easy to culture and stable in batch. Dry centella asiatica powder is taken as a raw material and fed into the culture medium for conversion, asiaticoside separation and extraction operations are omitted, separation and purification steps of enzyme are also omitted, the process is simple, and industrial application is facilitated; the content of the asiatic acid can be increased by 3 times at most after the centella asiatica powder is subjected to biotransformation treatment.

Description

(1) Technical field [0001] The invention relates to a glucosidase-producing Aspergillus niger and its application in biotransformation to increase the content of asiatic acid in Centella asiatica. (2) Background technology [0002] Centella asiatica [Centella asiatica (L.) Urb], also known as peppermint, Lianmocao, Bengdawan, Mandixiang, is a plant of the genus Centella asiatica in the family Umbelliferae. It has the effects of clearing away heat and dampness, detoxifying and reducing swelling. Our folks are commonly used to treat colds, tonsillitis, infectious hepatitis, dysentery, bruises, boils, swollen toxins, and traumatic bleeding. Centella asiatica contains triterpene saponins and flavonoids, among which the active pharmacological components are mainly saponins and their saponins, including asiaticoside, asiatic acid, madecassoside and madecassic acid, etc. Therefore, the extraction and purification of these substances has always been the focus of modern application ...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P33/00C12R1/685
CPCC12P33/00C12N1/145C12R2001/685
Inventor 梅建凤金航李靓陈虹应国清王鸿易喻
Owner ZHEJIANG UNIV OF TECH
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