A strain of Saccharomyces cerevisiae and its application in fermentative production of s-adenosylmethionine
A technology for adenosylmethionine and Saccharomyces cerevisiae strains, applied in the field of microorganisms, can solve the problems of affecting the yield of S-adenosylmethionine, increasing the burden of S-adenosylmethionine production condition control, and achieving simple and easy-to-control culture conditions and clear and simple process. Easy to control and good stability
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Embodiment 1
[0031]The inventor used low-energy ion beam implantation technology to screen the Saccharomyces cerevisiae that can obtain the product S-adenosylmethionine under the induction of L-methionine in the soil; at 30°C, the screened Saccharomyces cerevisiae Inoculate into YPD culture medium and cultivate for 4 hours, centrifuge to remove the supernatant, add appropriate amount of normal saline to make S. After dilution, the strained bacterial suspension was spread on the YPD solid medium for culture; a single strain with good growth was selected and inoculated in the liquid fermentation medium for culture, and after three rounds of screening, the stable genetic traits capable of high-yielding S-adenosine were selected. Saccharomyces cerevisiae strains of methionine, that is, Saccharomyces cerevisiae strains with high production of S-adenosylmethionine.
Embodiment 2
[0033] Plate medium: 20 g / L peptone, 10 g / L yeast extract, 20 g / L glucose, 20 g / L agar, natural pH.
[0034] Aerobic fermentation medium: glucose 37.5 g / L, corn steep liquor 45g / L, malt extract powder 15g / L, K 2 HPO 4 •3H 2 O1.6 g / L, KH 2 PO 4 1.25 g / L, MgSO 4 •7H 2 O 3.75 g / L, initial pH 5.0. Fill a 500mL Erlenmeyer flask with a volume of 30mL, and sterilize at 121°C for 15 minutes.
[0035] The initial strain and CGMCC No. 13760 bacteria were firstly activated on the plate at 30°C, and after 24 hours, two rings of well-grown bacteria were put into 30mL fermentation medium, cultured at 30°C, 200rpm for 12h, and then 3% (v / v ) the inoculum amount is transferred to a 30ml fermentation broth / 500ml Erlenmeyer flask, cultured at 30°C, and the rotation speed is 200rpm. After the bacteria grow to the mid-logarithmic growth phase (that is, fermentation for 48 hours), add 0.1 g of D / L-methionine to the fermentation broth. After 30 hours of fermentation, the fermentation broth...
Embodiment 3
[0037] Plate culture medium: with embodiment 1;
[0038] Aerobic fermentation medium: glucose 60g / L, corn steep liquor 10g / L, malt extract powder 5g / L, yeast powder 15g / L, acidified molasses 40 g / L, K 2 HPO 4 •3H 2 O1.6 g / L, KH 2 PO 4 1.25 g / L, MgSO 4 •7H 2 O 3.75 g / L, FeSO 4 •7H 2 O 10mg / L, (NH 4 ) 2 HPO 4 5 g / L, initial pH 5.0. 7 L fermenter with 3 L volume, sterilized at 121°C for 15 minutes.
[0039] The initial strain and CGMCC No.13760 bacteria were firstly activated on the plate at 30°C, and after 24 hours, two rings of well-grown bacteria were put into 30mL fermentation medium, cultivated at 30°C, 200rpm for 12h, and then according to 10% (v / v ) inoculum amount was transferred into 3L fermentation medium, ventilated with air at 2v / vm, rotated at 800rpm, and cultured at 30°C. After 16 hours of fermentation, start to add glucose, and control the concentration of glucose to 10-15g / L. After the bacteria grow to the mid-logarithmic growth stage (ie, 36 hours...
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