A mutant strain of Rhodobacter sphaeroides that produces coenzyme q10 and method for producing coenzyme q10 by fermentation
A technology of Rhodobacter sphaeroides and mutant strains, applied in the biological field, can solve the problem of low product yield and achieve the effect of increasing the fermentation unit
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Embodiment 1
[0055] The screening of embodiment 1 Rhodobacter sphaeroides mutant strain
[0056] Taking the original strain of Rhodobacter sphaeroides YLL-13 isolated from sewage treatment plant sludge as the starting strain to screen the mutant strain of Rhodobacter sphaeroides, the steps are as follows:
[0057] 1. Preparation of culture medium
[0058] 1) Solid medium: glucose 3g / L, yeast powder 8g / L, NaCl 2g / L, KH 2 PO 4 1.3g / L, MgSO 4 125mg / L, biotin 15μg / L, niacin 1mg / L, thiamine hydrochloride 1mg / L, agar powder 15g / L, pH 7.2.
[0059] 2) Seed medium: glucose 3g / L, yeast powder 8g / L, corn steep liquor powder 1.0g / L, NaCl 2.0g / L, KH 2 PO 4 1.3g / L, MgSO 4 125mg / L, FeSO 4 100mg / L, MnSO 4 1.0mg / L, biotin 15μg / L, niacin 1mg / L, thiamine hydrochloride 1mg / L, pH 7.2.
[0060] 2. The original strain YLL-13 was inoculated in the seed medium, and cultured at 32°C and 200r / min for 24h.
[0061] 3. Take 1mL of bacterial suspension, centrifuge at 5000r / min for 5min to obtain bacteria...
Embodiment 2
[0069] Example 2 YLL-13 Fermentation Process for Coenzyme Q10 Production
[0070] 1. Fermentation culture
[0071] Pick activated YLL-13 colonies that were cultured statically at 32°C for 4-5 days on a solid plate, inoculate them in a 250mL Erlenmeyer flask containing 50mL of seed medium, and culture them at 32°C and 200r / min for 24h; then Transfer all 50 mL of bacterial suspension to a 1L Erlenmeyer flask containing 450 mL of seed medium, and continue culturing for 120 h at 32°C and 200 r / min.
[0072] The composition of the seed medium is: glucose 3g / L, yeast powder 8g / L, corn steep liquor dry powder 1.0g / L, NaCl 2.0g / L, KH 2 PO 4 1.3g / L, MgSO 4 125mg / L, FeSO 4 100mg / L, MnSO 4 1.0mg / L, biotin 15μg / L, niacin 1mg / L, thiamine hydrochloride 1mg / L, pH 7.2.
[0073] 2. Extraction of coenzyme Q10
[0074] Take 1mL of fermentation bacteria suspension, add 200μL of 20mmol / L HCl and mix well, bathe in water at 75°C for 15min, centrifuge at 5000r / min for 10min, discard the s...
Embodiment 3
[0083] Example 3 YLL-13 fermentation production of coenzyme Q10 optimization process
[0084] 1. Preparation of culture medium
[0085] 1) Seed medium
[0086] Glucose 3g / L, yeast powder 8g / L, corn steep liquor dry powder 1.0g / L, NaCl 2.0g / L, KH 2 PO 4 1.3g / L, MgSO 4 125mg / L, FeSO 4 100mg / L, MnSO 4 1.0mg / L, biotin 15μg / L, niacin 1mg / L, thiamine hydrochloride 1mg / L, pH7.2.
[0087] 2) Fermentation medium
[0088] Glucose 40g / L, corn steep liquor dry powder 5.5g / L, sodium glutamate 3.0g / L, NaCl 2.0g / L, CaCl 2 0.1g / L, KH 2 PO 4 2.5g / L, MgSO 4 6.3g / L, NH 4 SO 4 2.5g / L, FeSO 4 100mg / L, MnSO 41mg / L, CoCl 2 20mg / L, biotin 15μg / L, niacin 1mg / L, thiamine hydrochloride 1mg / L, pH 7.2.
[0089] 2. Fermentation culture
[0090] 1) First class seeds
[0091] Activated YLL-13 colonies that were cultured at 32°C for 4-5 days on a solid plate were picked, inoculated into a 250mL conical flask containing 50mL of seed medium, and cultured at 32°C and 200r / min for 24h. ...
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