Application of endophytic cladosporium fungus of liriope spicata var.prolifera Y.T.MA in preparing steroid saponin
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A technology of steroidal saponins and Cladosporium, applied in the field of microorganisms, can solve the problems of long production cycle of tubers, low content of steroidal saponins, and high cost of saponin extraction
Active Publication Date: 2018-02-23
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 through the separation of root extracts
However, in the actual production of Ophiopogon japonicus, there are pr
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Embodiment 1
[0048] The living root tubers of Xiangmaidong used for the isolation of the endophytic fungus EF111 strain of the present invention come 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 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 then dip ...
Embodiment 2
[0055] Utilize above-mentioned endophytic fungus to prepare the step of Ophiopogon steroidal saponin as follows:
[0056] (1) Take the endophytic fungus EF111 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) By 200g potatoes, 20g glucose, the proportioning of 1000ml distilled water prepares the liquid fermentation culture medium, respectively packs in the Erlenmeyer flask of 250ml, and every bottle is about 100ml, sterilizes, and cools for standby; Under aseptic conditions, according to 10 % of the inoculum was inserted into the seeds, and cultured with ...
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Abstract
The invention provides application of endophytic cladosporium fungus of liriope spicata var.prolifera Y.T.MA in preparing steroid saponin, and relates to the technical field of microorganisms. The application is mainly used for solving the problems that most existing steroid saponin of liriope spicata var.prolifera Y.T.MA is from extracts of liriope spicata var.prolifera Y.T.MA, tubers of liriopespicata var.prolifera Y.T.MA are long in production period, low in steroid saponin content and high in saponin extraction cost, and accordingly the market volume of the steroid saponin of liriope spicata var.prolifera Y.T.MA is limited. The fungus is separated from living tubers of liriope spicata var.prolifera Y.T.MA, authenticated as Cladosporium sp.EF111 by means of the 18 S rDNA sequence, andpreserved in the China Center for Type Culture Collection. The preservation date is December 7th, 2016, and the preservation number of CCTCC No:M2016725. According to the application, liriope spicatavar.prolifera Y.T.MA produced by a liriope spicata var.prolifera Y.T.MA GAP base which is the original production place of liriope spicata var.prolifera Y.T.MA in Oumiao town, Xiangyang city in Hubeiprovince, endophytic fungus strain liquid of liriope spicata var.prolifera Y.T.MA is used for fermentation, and the steroid saponin of liriope spicata var.prolifera Y.T.MA is produced. The endophyticfungus is an important microorganism for finding new resources of steroid saponin of liriope spicata var.prolifera Y.T.MA.
Description
technical field [0001] The invention relates to the technical field of microorganisms, in particular to an endophytic fungus of Ophiopogon japonicus, and the endophytic fungus is a fungus of the genus Cladosporium 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...
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