Method for producing L-lactic acid by utilizing corn cob and special rhizopus oryzae thereof
A corncob, lactic acid technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of high nutritional requirements, reduced production costs, and high raw material costs
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Embodiment 1
[0025] Embodiment 1, using glucose as raw material to ferment and produce L-lactic acid
[0026] 1. Isolation and purification of strains
[0027] 1) Enrichment and separation
[0028] The rice was placed in a petri dish covered with two layers of wet filter paper (the filter paper was sprayed with 1% lactic acid by mass percentage), and then the surface of the rice was sprayed with 1% lactic acid by mass percentage. Take 1g of soil sample (taken near the dairy farm in Yuncheng, Shanxi) and sprinkle it on the surface of the rice, cover it with a lid, and incubate at 30°C for 3-4 days.
[0029] 2) Plate separation and purification
[0030] Pick a single colony from a petri dish and inoculate it in the primary screening medium for single colony isolation, and culture at 30°C for 3 to 4 days. A single colony producing a larger yellow color change circle was transferred to PDA medium and cultured under the same conditions to obtain Rhizopus oryzae (Rhizopus oryzae) GY-18.
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Embodiment 2
[0044] Embodiment 2, take xylose as raw material fermentation production lactic acid
[0045] Preparation of fermentation medium: xylose was 20, 40, 60, 80 and 100g, NH 4 Cl2.0g, KH 2 PO 4 0.3g, MgSO 4 ·7H 2 O0.25g, ZnSO 4 ·7H 2 O0.08g, CaCO 3 40g, add water to 1000mL, adjust pH to 6.0, and autoclave at 115°C for 30min for later use.
[0046] The rhizopus oryzae GY-18CGMCC2681 obtained by the screening of the above-mentioned embodiment 1 step 1 is connected on the PDA medium, cultivated for 2-3 days in a 25-35° C. of glycerol to elute the spores to make 1 x 10 5 -1×10 8 / mL of spore suspension. Take 10 mL of the above spore suspension and inoculate it into 1000 mL of the above fermentation medium, culture it at 180 rpm (rotation radius: 15 mm) at 35°C for 120 h, and use an equal volume of 1 mol / L sulfuric acid solution to precipitate calcium ions in the obtained fermentation broth, and centrifuge at 12000 rpm for 10 min , take the supernatant, which is the fermentat...
Embodiment 3
[0050] Embodiment 3, take glucose and xylose as raw material fermentation production lactic acid
[0051] Preparation of fermentation medium: glucose 40g, xylose 40g, NH 4 Cl2.0g, KH 2 PO 4 0.3g, MgSO 4 ·7H 2 O0.25g, ZnSO 4 ·7H 2 O0.08g, CaCO 3 40g, add water to 1000mL, adjust pH to 6.0, and autoclave at 115°C for 30min for later use.
[0052] The rhizopus oryzae GY-18CGMCC2681 obtained by the screening of the above-mentioned embodiment 1 step 1 is connected on the PDA medium, cultivated for 2-3 days in a 25-35° C. of glycerol to elute the spores to make 1 x 10 5 -1×10 8 / mL of spore suspension. Take 20mL of the above spore suspension and inoculate it into 1000mL of the above fermentation medium, culture it at 180rpm (radius of rotation: 15mm) at 35°C for 120h, and use an equal volume of 1mol / L sulfuric acid solution to precipitate calcium ions in the obtained fermentation broth, and centrifuge at 12000rpm for 10min , take the supernatant, which is the fermentation ...
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