Bacillus subtilis SWB8 for producing diosgenin

A technology of Bacillus subtilis and diosgenin, applied in the field of applied microorganisms, can solve problems such as environmental pollution and backward technology, and achieve the effect of avoiding environmental pollution and stabilizing capacity

Inactive Publication Date: 2011-05-18
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Domestically, strong acid hydrolysis and other technologies are mainly used to extract diosgenin from turmeric plants, but the technology is backward. Statistics show that 140 tons of fresh turmeric can be processed by traditional strong acid hydrolysis, which can produce 1 ton of saponin and produce 400-500 tons of waste water, COD 30000mg / L, BOD 8000mg / L, ammonia nitrogen 300 mg / L and 10 tons of waste residue (Shen Kangrong et al., Turmeric Development and Planting Technology, Hubei Science and Technology Press), causing serious pollution to the environment

Method used

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  • Bacillus subtilis SWB8 for producing diosgenin
  • Bacillus subtilis SWB8 for producing diosgenin
  • Bacillus subtilis SWB8 for producing diosgenin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment one. Bacillus subtilis SWB8 ( Bacillus subtilis SWB8) Isolation and identification of CCTCC M 2010271

[0026] 1. Isolation and purification of strains

[0027] (1) Separation. Medium: 0.3% beef extract; 1% peptone; 0.5% NaCl; 2% agar; 1% turmeric dry powder (the turmeric is dried and crushed into powder after removing the brown skin); distilled water; pH7.0. Select the underground stems of the fresh two-year-old turmeric plant, wash them, burn them with 95% alcohol, peel off the brown skin under aseptic conditions, soak them in 70% alcohol for 10 minutes, wash them with sterile distilled water, and then peel off the outer layer tissue with a thickness of 2-3mm. The internal tissues were cut into 2-3 mm thick slices and pasted on the culture medium, and cultured at a constant temperature at 32°C.

[0028](2) Purification. Beef extract peptone medium: 0.3% beef extract; 1% peptone; 0.5% NaCl; 2% agar; 1% turmeric dry powder; distilled water; pH7.0. Czap...

Embodiment 2

[0039] Embodiment two. Bacillus subtilis SWB8 ( Bacillus subtilis SWB8) CCTCC M 2010271 Diosgenin produced by liquid fermentation

[0040] The standard strains of Bacillus megaterium SWB15 (an endophyte isolated from turmeric) and Bacillus subtilis (ATCC6633) were selected as the control strains of Bacillus subtilis SWB8, and were fermented under the same conditions, and the fermentation products were extracted for the detection of diosgenin.

[0041] 1. Strain activation

[0042] Pick and preserve the strains, dilute and mix them with normal saline, spread them on the seed solid medium (0.3% beef extract; 1% peptone; 0.5% NaCl; 2% agar; distilled water; pH7.0), 30~35℃, Cultivate at constant temperature for 48-72 hours. Under the same conditions, replant once to rejuvenate the strain.

[0043] 2. Liquid fermentation

[0044] Pick the bacterial strains on the seed medium and dilute and mix with normal saline, draw 10 5 Inoculate the bacterial liquid of each cell into 100...

Embodiment 3

[0051] Embodiment three. Bacillus subtilis SWB8 ( Bacillus subtilis SWB8) Antibacterial activity of diosgenin produced by CCTCC M 2010271

[0052] Pick the test strains (Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Salmonella typhi, Salmonella paratyphi A, Shigella dysenteriae) diluted with normal saline and spread on corresponding common peptone solids On the culture medium, culture at 37°C for 48h. Using dimethyl sulfoxide (DMSO) as a solvent, a certain concentration of Bacillus subtilis SWB8 ( Bacillus subtilis The detection solution of diosgenin produced by SWB8) absorbs the The paper sheet of the detection solution is pasted on the coated with 10 7 ~10 8 Incubate at 37°C for 24 hours on the solid medium of each test bacterium with the number of bacterium, and measure the diameter of the inhibition zone. DMSO solution was set as the control in the experiment. The results showed that Bacillus subtilis SWB8 ( Bacillus subtilis SWB8) fermented di...

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Abstract

The invention discloses a Bacillus subtilis WB8 capable of producing diosgenin, and belongs to the technical field of applied microbiology. The strain is separated from internal issues of subterraneous stems of a medicinal plant of Dioscorea zingiberensis wright, can synthesize and secrete a metabolin of a host plant, namely diosgenin, and has the preservation number of CCTCC (China Center For Type Culture Collection) NO:M2010271; and the strain is rod-like, has the size of 1-1.5*3-5 mu m, forms a single-chain sporocarp, releases endospore, has rough colony with grid bulges, forms pellicle through liquid fermentation, has rich foam, and can grow 16SrDNA core sequence 1542bp with the thermophilic temperature of between 30 and 35 DEG C, the pH of 6.0-9.0 and capacity of tolerating 9.0 percent NaCl, wherein the core sequence has the sequence number of HM210636. The strain liquid is fermented by a common culture medium for 48 to 60 hours, the chloroform extract of the fermentation liquor is the diosgenin, and the diosgenin has the activity of inhibiting pathogenic bacteria and resisting tumor cells. The strain provided by the invention can produce the diosgenin; and compared with the conventional process of extracting the diosgenin from yam plants, the method avoids the pollution of strong acid and yam byproducts on environment, and has potential industrial application prospect.

Description

technical field [0001] The invention relates to the technical field of applied microorganisms, in particular to an endogenous Bacillus subtilis SWB8 ( Bacillus subtilis SWB8). Background technique [0002] Diosgenin (Diosgenin, also known as saponin) is the hydrolyzed product of diosgenin, which is mainly used as a raw material for the synthesis of steroidal drugs, and has been widely used in cosmetics, health care, contraceptive analgesia, anesthesia, lowering cholesterol and treating diabetes, etc. drug. At present, diosgenin is mainly derived from the genus Dioscorea ( Dioscorea ), Fenugreek ( Trigonella ) and Zingiber officinalis ( Costus )plant. Dioscorea shield leaf ( Dioscorea zingiberensis C.H. Wright), also known as turmeric, is a perennial herbaceous vine, and its underground stem contains 45-50% starch, 40-50% cellulose and 2-3% diosgenin. In China, turmeric is the main source of extracting diosgenin, with a planting area of ​​1.8 million mu at its p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P17/04A61K31/58A61P31/04A61P35/00C12R1/125
Inventor 金志雄万永继周围王晓强冯伟高红张冉汪静杰
Owner SOUTHWEST UNIVERSITY
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