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A technique for producing adenine and bacteria, which is applied in the field of fermentation and production of adenine, and can solve problems such as difficulty in extracting adenine
Pending Publication Date: 2022-07-29
TIANJIN UNIV OF SCI & TECH
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The current production method of adenine is to produce adenine by enzymatically hydrolyzing adenosine. Using this method to produce adenine will produce a large amount of D-ribose, which will cause great difficulties in the extraction of adenine in the later stage.
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Embodiment 1
[0031] A method for producing adenine by fermentation, the concrete steps are as follows:
[0032] 1. Bacillus subtilis XGL culture
[0033] (1) Activation of strains: Bacillus subtilis XGL was inserted into the slanted medium from the glycerol storage tube for activation and cultivation (two generations of activation), and the temperature was maintained at 34 ° C; the composition and content of the slanted medium were: glucose 2g / L, beef extract 10g / L, sodium chloride 2.5g / L, peptone 10g / L, yeast powder 2g / L, agar powder 25g / L, xanthine 50mg / L, histidine 50mg / L;
[0034] (2) Seed culture (2L): Pour about 200 mL of sterile water into the above-mentioned eggplant-shaped bottle near the flame of the ultra-clean bench, scrape the colonies into sterile water with an inoculating ring and disperse to prepare a bacterial suspension, and place in a fermenter. The bacterial suspension is inoculated into the fermented seed culture tank beside the fire ring. During the cultivation proc...
Embodiment 2
[0046] A method for producing adenine by fermentation, with reference to Example 1, the difference is: the time of three mixing operations is selected as 35h, 55h and 75h respectively;
[0047] When the fermentation was put in the tank for 85 hours, the total adenosine amount was 132 g, the total adenine production was 47 g, and the total D-ribose production was 0 g.
Embodiment 3
[0049] A method for producing adenine by fermentation, with reference to Example 1, the difference is: the time of the three mixing operations is selected as 40h, 60h and 80h respectively;
[0050] When the fermentation was put in the tank for 85 hours, the total adenosine amount was 138 g, the total adenine production was 45 g, and the total D-ribose production was 0 g.
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Abstract
The invention provides a method for producing adenine by fermentation, and the adenine is produced by adopting a mixed fermentation method, so that D-ribose generated after adenosine enzymolysis can be reutilized by bacteria, the consumption of sugar after bacteria mixing is reduced, the later extraction difficulty of adenine is also reduced, and the yield of adenine is improved. Meanwhile, the BL21-ADH thallus culture solution subjected to ultrasonic treatment not only can retain the activity of adenosine hydrolase, but also can further improve the thallus activity of the bacillus subtilis XGL in the later fermentation period after being subjected to ultrasonic crushing, and meanwhile, the thallus activity of the bacillus subtilis XGL in the later fermentation period is further improved by dialyzing the thallus liquid in the later fermentation period through a dialysis fermentation method; the living environment of the thalli is purified; meanwhile, the feedback inhibition of the final product on the thalli is relieved.
Description
technical field [0001] The invention relates to the technical field of microorganisms and fermentation engineering, in particular to a method for producing adenine by fermentation. Background technique [0002] Adenine, also known as 6-aminopurine, is a base in deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), abbreviated as A. In DNA, it pairs with thymine (T). In ribonucleic acid, it is paired with uracil (U). Formerly known as vitamin B4. Adenine also has many different biochemical functions. In cellular respiration, it acts in the form of energy-rich adenosine triphosphate (ATP), as well as cofactors Chemicalbook nicotinamide adenine dinucleotide (NAD) and flavin adenine dinucleotide (FAD); In the process of protein biosynthesis as a component of DNA and RNA. The current production method of adenine is to produce adenine by enzymatic hydrolysis of adenosine. Using this method to produce adenine will produce a large amount of D-ribose, which causes great diffic...
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