Liriope spicatavar endogenous aspergillus niger and application thereof in preparing steroid saponin
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A technology of Aspergillus niger and steroidal saponins, which is applied in the field of microorganisms and can solve the problems of long root production cycle, low content of steroidal saponins, and high cost of saponin extraction
Active Publication Date: 2018-04-24
HUBEI UNIV OF ARTS & SCI
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[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 by separating 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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Embodiment 1
[0048] The Xiangmaidong live tuber used for the isolation of the Xiangmaidong endophytic fungus EF026 strain of the present invention comes from the Xiangmaidong GAP base, the place of origin of Xiangmaidong in Oumiao Town, Xiangyang City, Hubei Province.
[0049] Xiangmaidong 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...
Embodiment 2
[0055] The steps of preparing Xiangmaidong steroidal saponins by utilizing the above-mentioned Xiangmaidong endophytic fungus:
[0056] (1) Take the endophytic fungus EF026 strain 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, activate and cultivate for 72 hours at 28°C;
[0057] (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;
[0058] (3) Put the prepared liquid fermentation medium into 250ml Erlenmeyer flasks, about 100ml per bottle, sterilize and cool for later use; 180rpm shaking culture for 7 days;
[0059] (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 st...
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Abstract
The invention discloses liriope spicatavar endogenous aspergillus niger and application thereof in preparing steroid saponin, relates to the technical field of microorganisms and mainly solves the problem that liriope spicatavar steroid saponin market release quantity is limited caused by the fact that liriope spicatavar is long in tuberous root production period, low in steroid saponin content and high in saponin extraction cost because most liriope spicatavar steroid saponin directly comes from liriope spicatavar extract at present. Endogenous fungus is separated from living tuberous root ofLiriope spicatavar. Prolifera Y. T. Ma, identified as Aspergillus niger through 18S rDNA sequence and already collected at the China center for Type Culture Collection on Dec. 15, 2016 under the number of CCTCC No:M2016753. Liriope spicatavar produced by a liriope spicatavar GAP base which is a liriope spicatavar origin in Oumiao Town, Xiangyang City of Hubei Province is utilized, and liriope spicatavar steroid saponin is generated through liquid fermentation of liriope spicatavar endogenous fungus strain. The endogenous fungus is an important microorganism for searching new resource of liriope spicatavar steroid saponin.
Description
technical field [0001] The invention relates to the technical field of microorganisms, in particular to an endophytic fungus of Xiangmaidong, which is an endophytic Aspergillus niger capable of producing Xiangmaidong steroidal saponins. 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 ring) four types. Now 14 kinds of saponins...
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