Virus-free in situ and alcohol fermentation method of composite bacteria for lignocellulose hydrolysis product
A technology of lignocellulose and hydrolyzate, which is applied in the field of in-situ detoxification ethanol fermentation, can solve the problems such as the complexity and cost of ethanol fermentation process, and achieve the effects of good mixed fermentation compatibility, convenient operation and fast fermentation speed
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Embodiment 1
[0038] Example 1 In-situ detoxification ethanol fermentation of lignocellulose hydrolyzate by composite bacteria of the present invention
[0039] The strains S.cerevisiaeY5 (Y5) and I.orientalis Y4 (Y4) were screened and preserved by our laboratory. The strain P.stipitis CBS6054 (CBS6054) was purchased from ATCC, USA.
[0040] Preparation of proliferation medium: glucose 20g / L, yeast extract 10g / L, peptone 20g / L, pH5.0-5.5, sterilized at 121°C for 20min.
[0041] Preparation of fermentation medium: yeast extract 10g / L, peptone 20g / L, lignocellulose dilute acid hydrolyzate 100ml (wherein glucose is 15.4g / L, xylose 6.6g / L, furfural 1.37g / L, 5-HMF0 .47g / L, acetic acid 10g / LCa(OH) 2 Adjust the pH of the hydrolyzate to 5.0), and sterilize at 115°C for 30min.
[0042] Inoculate strains Y5 and CBS6054 into 100ml proliferation medium, culture at 30°C and 150rpm for 12-14h, and the viable counts of the obtained strains were 10 7 cfu / L, 10 8 cfu / L.
[0043] Inoculate 5ml of CBS60...
experiment example 1
[0044] Ethanol fermentation of experimental example 1 single strain Y5, Y4, CBS6054
[0045] 1. Test material
[0046] The strains S.cerevisiaeY5 (Y5) and I.orientalis Y4 (Y4) were screened and preserved by our laboratory. The strain P.stipitis CBS6054 (CBS6054) was purchased from ATCC, USA.
[0047] 2. Test method
[0048]The bacterial strains S.cerevisiaeY5, I.orientalisY4 and P.stipitisCBS6054 were respectively inoculated in 100ml of the proliferation medium in Example 1, cultured at 30°C and 150rpm for 12h, and the dry cell weights of the obtained strains were 2.7g / L and 3.0g / L respectively. g / L, 3.3g / L.
[0049] Take 5ml each of Y5, Y4 and CBS6054 bacteria solution and inoculate them into 100ml of glucose, glucose+, xylose, xylose+, mixed sugar, mixed sugar+ (the concentrations of glucose and xylose are 24g / l and 13g / l respectively) In the liquid culture medium, "+" means that furfural 1.37g / L and 5-HMF0.47g / L were added, cultured at 30°C and 80rpm, and the concentrat...
experiment example 2
[0064] Experimental Example 2 Mixed sugar ethanol fermentation of mixed strains Y5+CBS6054 and Y4+CBS6054
[0065] 1. Test material
[0066] Bacteria and proliferation medium are the same as in Experimental Example 1.
[0067] Preparation of fermentation medium: yeast extract 10g / L, peptone 20g / L, glucose 24g / L, xylose 13g / L, furfural 1.37g / L, 5-HMF0.47g / L, adjust the pH of the hydrolyzate with calcium hydroxide To 5.0, sterilize at 115°C for 30min.
[0068] 2. Test method
[0069] 5ml of CBS6054 and Y5 seed solutions were inoculated in 100ml fermentation medium, 5ml of CBS6054 and Y4 seed solutions were inoculated in 100ml fermentation medium, cultured at 30°C and 80rpm, and the concentrations of ethanol, sugar, furfural and 5-HMF were measured .
[0070] 3. Analytical method (with experimental example 1)
[0071] 4. Calculation method (same as experimental example 1)
[0072] 5. Test results
[0073] (1) Mixed sugar ethanol fermentation of compound bacteria Y5+CBS6054...
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