A Corynebacterium glutamicum producing high citrulline and its application in fermentative production of citrulline
A technology of Corynebacterium glutamicum and citrulline, applied in the field of microorganisms, can solve the problems of affecting the quality of fermentation products and increasing the separation load, and achieve the effect of easy process, process control and single fermentation product
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Embodiment 1
[0030] Plate medium: peptone 10 g / L, yeast extract 5 g / L, NaCl 10 g / L, agar 20 g / L, pH7.0.
[0031] Aerobic fermentation medium: glucose 40g / L, K 2 HPO 4 •3H 2 O 1.5g / L, KH 2 PO 4 0.5g / L, MgSO 4 •7H 2 O0.4 g / L, FeSO 4 •7H 2 O 20mg / L, urea 5g / L, biotin 100μg / L, vitamin B1 200μg / L, initial pH 7.0. 500mL Erlenmeyer bottle with 30mL volume, sterilized at 115°C for 10 minutes.
[0032] The bacteria with the preservation numbers ATCC13032 and CGMCC No.11567 were first 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 and 200rpm for 12h, and then according to 3% (v / v) The amount of inoculum is transferred to a 30ml fermentation broth / 500ml Erlenmeyer flask, cultured at 30°C, and the rotation speed is 200rpm. After 48 hours of fermentation, the fermentation broth of CGMCCNo.11567 contained 5.43g / L citrulline. Compared with ATCC13032, the yield of citrulline is increased by 10.24 tim...
Embodiment 2
[0034] Plate culture medium: with embodiment 1;
[0035] Aerobic fermentation medium: corn mash liquid 500mL / L (the sugar content of corn mash liquid is about 100g / L), K 2 HPO 4 •3H 2 O 1.5g / L, KH 2 PO 4 0.5g / L, MgSO 4 •7H 2 O 0.4 g / L, FeSO 4 •7H 2 O 20mg / L, urea 2.5g / L, biotin 100μg / L, vitamin B1 200μg / L, initial pH 7.0. 5L fermenter with 4L liquid, sterilized at 115°C for 10 minutes.
[0036] The bacteria with the preservation numbers ATCC13032 and CGMCC No.11567 were first 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 and 200rpm for 12h, and then according to 3% (v / v) Transfer the inoculum amount into 4L of fermentation medium, ventilate with air at 1v / vm, rotate at 800rpm, and culture at 30°C. After 60 hours of fermentation, the fermentation broth of CGMCC No. contained 12.8g / L citrulline. Compared with ATCC13032, the yield of citrulline is increased by 24.2 times,...
Embodiment 3
[0038] Plate culture medium: with embodiment 1;
[0039]Aerobic fermentation medium: sucrose 40g / L, corn steep liquor dry powder 10g / L, initial pH 7.0. 5L fermentation tank contains 4L liquid, sterilized at 115°C for 10 minutes.
[0040] The bacteria with the preservation numbers ATCC13032 and CGMCC No.11567 were first 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 and 200rpm for 12h, and then according to 3% (v / v) Transfer the inoculum amount into 4L of fermentation medium, ventilate with air at 1v / vm, rotate at 800rpm, and culture at 30°C. After 60 hours of fermentation, the fermentation broth of CGMCC No.11567 contained 11.8g / L citrulline. Compared with ATCC13032, the yield of citrulline is increased by 22.2 times, and there is no by-product L-lysine and L-glutamic acid.
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