Fermentation method for producing gamatine
A technology of agmatine and fermentation liquid, applied in the field of bioengineering, can solve the problems of low yield, pollution, and many steps in the chemical method route, and achieve the effect of ensuring food safety and broad industrial application prospects
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Embodiment 1
[0041] Example 1 : Preculture and fermentation
[0042] The bacterial strain CGMCC No.12601 preservation solution packed in the glycerol tube was thawed, and inoculated in the seed medium (glucose 75g / L, corn steep liquor 25-30g / L, (NH4) 2 SO 4 20g / L, KH 2 PO 4 1.5g / L, MgSO 4 ·7H 2 O 0.5g / L, urea 1.0g / L, histidine 30mg / L, molasses 25g / L, biotin 100μg / L, pH 7.0), 5000mL Erlenmeyer bottle liquid volume 500mL, 30 ℃ for 18 hours, absorbance value When ΔOD=0.4-0.5, shake the bed to obtain seed culture solution. After testing, no bacteria.
[0043] Inoculate 500 mL of seed culture solution into a 7-liter fermenter with a liquid volume of 5 liters. The composition of the medium is the same as that of the seed medium, pH=6.4-6.7 (after sterilization). Feed medium: ammonium sulfate 500g / L, glucose 650g / L. The culture temperature is 30°C, the tank pressure is 0.05Mpa, and the initial aeration ratio is 1vvm. Stirring 600rpm. During the fermentation, the pH is controlled at...
Embodiment 2
[0044] Example 2 : Separation of fermentation broth
[0045] 5.2 liters of fermented broth obtained in Example 1 was centrifuged at 4500 rpm, and the thallus was discarded. Add 1% activated carbon to the supernatant, stir and decolorize for 30 minutes, then add 1% diatomaceous earth and filter to obtain a clear filtrate.
Embodiment 3
[0046] Example 3 : Extraction of Agmatine
[0047] The filtrate obtained in Example 2 was passed through strongly acidic cation exchange resin 732 at a rate of 1 column volume per hour, and agmatine was exchanged and adsorbed on the resin column. After washing with pure water for one column volume, 5% sulfuric acid was used for elution, and the eluate was collected.
[0048] The eluate was adjusted to pH=6.0 with ammonia water, 1% activated carbon was added, stirred for 30 minutes to decolorize, and suction filtered. The filtrate was collected and concentrated under reduced pressure. When concentrated to about 800g / L, it was cooled to room temperature to crystallize for more than 4 hours, and the crystals were collected by suction filtration to obtain 347 grams of wet crystals of agmatine sulfate.
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