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A kind of endophytic Penicillium oxalicum and its application in the preparation of steroidal saponins

A technology of Penicillium oxalicum and steroidal saponins, which is applied in the field of microorganisms and can solve the problems of long production cycle of tubers, environmental pollution, and restrictions on the market launch of Ophiopogon japonicus steroidal saponins.

Active Publication Date: 2019-12-27
HUBEI UNIV OF ARTS & SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Although with the progress of chiral synthesis and oligosaccharide synthesis methods, there have been some reports on the total synthesis of steroidal saponin compounds, but due to the complexity of the steroidal saponin structure, the synthesis is very difficult, and the chemical synthesis is harmful to the environment. Huge pollution, currently mainly obtained directly through the separation of root extracts
However, in the actual production of Ophiopogon japonicus, there are problems such as "small planting area of ​​continuous cropping obstacles", "greatly affected by climatic conditions", "long production cycle of roots", "low content of steroidal saponins", "high cost of saponin extraction". " and other factors have led to restrictions on the market release of xiangmaidong steroidal saponins
At present, there is no report about a kind of endophytic fungus capable of producing xiangmaidong steroidal saponins through microbial fermentation and its application

Method used

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  • A kind of endophytic Penicillium oxalicum and its application in the preparation of steroidal saponins
  • A kind of endophytic Penicillium oxalicum and its application in the preparation of steroidal saponins

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Embodiment 1

[0048] The Xiangmaidong living tuber used for the isolation of the Xiangmaidong endophytic fungus EF036 strain of the present invention comes from Xiangmaidong GAP base, the place of origin of Xiangmaidong in Oumiao Town, Xiangyang City, Hubei Province.

[0049] The endophytic fungus of the present invention is separated and obtained according to the following steps:

[0050] (1) Use a large amount of tap water to fully clean the soil on the surface of the root tuber, then put it into a clean beaker, add deionized water, and place it in an ultrasonic cleaner for repeated cleaning until the water after cleaning becomes extremely clear.

[0051] (2) After dipping the moisture on the surface of the tuber with sterile filter paper, proceed as follows: first soak the tuber with 75% (V / V) ethanol for 2 minutes, rinse it with sterile water for 3 times; Soak in -6% sodium hypochlorite solution for 2 minutes, then rinse continuously with a large amount of sterile water for 4 times, and...

Embodiment 2

[0055] (1) Take the endophytic fungus EF036 bacterial classification of the present invention, under aseptic conditions, pick a small amount of hyphae with an inoculation needle, insert into a sterilized PDA medium test tube, and activate and cultivate for 72 hours at 28°C;

[0056] (2) Take the strains after the activated culture, transfer them into the sterilized liquid potato seed culture medium under aseptic conditions, and shake and cultivate them at 180 rpm for 72 hours at 28° C. to obtain seeds;

[0057] (3) The prepared described liquid fermentation medium is packed into 250ml triangular flasks respectively, about 100ml of each bottle, sterilized, and cooled for standby; under aseptic conditions, insert seeds according to 10% inoculum size, 28 Cultivate with shaking at 180rpm for 7 days at ℃;

[0058] (4) After the fermentation is completed, the fermented solid-liquid mixture is placed in an ultra-low temperature refrigerator and quickly frozen into a solid state, and ...

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Abstract

The name of the invention is an endophytic Penicillium oxalate from Ophiopogon japonicus and its application in preparing steroidal saponins, and relates to the technical field of microorganisms. It is mainly to solve the problem that most of the steroidal saponins of Ophiopogon japonicus come directly from the extract of Ophiopogon japonicus. Due to the "long production cycle of root tubers", "low content of steroidal saponins" and "high cost of saponin extraction", Ophiopogon japonicus The problem of limited availability of winter steroidal saponins in the market. Penicillium oxalate of the present invention is isolated from Ophiopogon japonicus ( Liriope spicata var. prolifera Y.T.Ma), identified by 18S rDNA sequence as Penicillium oxalicum ( Penicillium oxalicum) , has been deposited in the China Center for Type Culture Collection, the preservation date is: December 7, 2016, and the preservation number is CCTCC No. M2016722. The present invention utilizes the Ophiopogon japonicus produced from the origin of Ophiopogon japonicus GAP base in Oumiao Town, Xiangyang City, Hubei Province, and produces Ophiopogon japonicus steroidal saponins through liquid fermentation of endophytic fungal strains of Opiocarpus japonicus; The endophytic fungi of the present invention are important microorganisms for finding new resources of Ophiopogon japonicus steroidal saponins.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to an endophytic fungus of Ophiopogon japonicus. The endophytic fungus is an endophytic Penicillium oxalicum capable of producing steroidal saponins from Ophiopogon japonicus. Background technique [0002] Xiangmaidong steroidal saponins are national geographical indication products, authentic Chinese medicinal plants in Hubei——Xiangmaidong Liriope spicatavar. prolifera The main active ingredient of Y. T. Ma tuber root, its saponin basic skeleton belongs to the derivative of Spirostane (Spirostane, which has 27 carbon atoms in total), according to the configuration of C25 and the ring closure state of the ring in the Spirostane structure, it can be It is divided into spirostanols (C25 is S configuration), isospirostanols (C25 is R configuration), furostanols (F ring is an open-chain derivative), deformed spirostanols Alcohols (F ring is a five-membered tetrahydrofuran rin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12N1/02C12P33/20C12R1/80
CPCC12N1/02C12N1/14C12P33/20C12N1/145C12R2001/80
Inventor 余海忠叶国栋潘婉莲黄立明
Owner HUBEI UNIV OF ARTS & SCI
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