Method for preparing 2,3-butanediol by using lemna minor as raw material
A butanediol and raw material technology, applied in the field of environmental biotechnology and biomass energy, can solve problems such as non-existence, achieve the effect of alleviating tension, alleviating food shortage, and solving secondary pollution
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[0012] Preparation of duckweed starch hydrolyzate: wash and dry the duckweed, then crush it through a 60-mesh sieve, use sterile water to prepare it into a suspension at a volume ratio of 1:8, and then add medium-temperature α-starch to it Enzymes and 0.2% CaCl 2 , liquefied at 60°C for several minutes, during which the starch degradation effect was detected with iodine solution until the color changed from blue to brown orange. Then the excessive α-amylase is inactivated in a boiling water bath state.
[0013] After it is cooled, add glucoamylase to it at a pH of about 4.5 to fully degrade it into monosaccharides, then inactivate the excess glucoamylase in a boiling water bath, and finally use 3,5-dinitro Salicylic acid method was used to determine the content of reducing sugar (for specific methods, refer to Chen Yuquan, editor-in-chief. Biochemical Experimental Methods and Techniques, Beijing: Science Press, 2002). Its principle is to utilize reducing sugar to have oxidat...
Embodiment 1
[0018] A 250ml Erlenmeyer flask was used for shake-flask fermentation. The total volume of the fermentation medium is 60ml, wherein the mass volume ratio of each component is as follows: duckweed starch hydrolyzate total sugar concentration 80.0g / L, yeast extract 6.0g / L, (NH 4 ) 2 HPO 4 1.0g / L, KCl0.2g / L, MgSO 4 ·7H 2 O0.3g / L, CaCO 3 3g / L, pH6.0. The above culture medium was sterilized under high pressure at 115°C for 30 minutes for later use. Then press the inoculum amount of 8% (V:V) into the liquid seeds, first ferment for 24 hours at 30°C and 200r / min, then adjust the rotating speed at 160r / min under the condition of constant temperature, and ferment for 24 hours to facilitate 2 , the synthesis of 3-butanediol, and finally the content of 2,3-butanediol was detected by high performance liquid chromatography.
Embodiment 2
[0020] Take 8g duckweed powder (starch content 30%, protein content 15%), 3.6g yeast extract, 6.0g (NH 4 ) 2 HPO 4 , 1.2g KCl, 1.8gMgSO 4 ·7H2 O, add 48.4ml of distilled water, place in a 250ml Erlenmeyer flask, adjust the pH value to 6.0 with hydrochloric acid and sodium hydroxide, and add CaCl with a mass volume concentration of 0.2% in the prepared liquid medium 2 In order to ensure the effect of enzymatic hydrolysis, starch was saccharified with medium-temperature α-amylase and glucoamylase respectively, sterilized at 115°C for 30 minutes, and finally CaCO 3 4.8g, then the seed solution was inoculated into the fermentation medium by the inoculum size of 6% (V:V), and after 60h of fermentation under 30°C and 160r / min conditions, the type of the fermentation product was detected by high performance liquid chromatography and Quantify it.
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