能源型甜菜品种的筛选及鉴定技术研究
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摘要
目前,能源问题威胁着各国的经济发展,影响着人类的生活。为了解决能源问题,世界各国均开展了对燃料乙醇的研究,其中微生物发酵法生产燃料乙醇已经成为国内外燃料乙醇研究和规模开发的热点。采用甜菜为原料发酵生产燃料乙醇具有种植面积广、发酵工艺简单、乙醇产量较高等优势。近几年一些国家开展了对能源甜菜的研究,但是没有一套完整的能源甜菜经济指标的鉴定技术。本论文通过优化酵母菌1373的发酵条件,对能源型甜菜甜菜品种进行筛选,并总结出能源甜菜经济指标的鉴定技术。
本试验分为六个部分。第一部分为田间试验。第二部分为最适菌株的选择试验,主要比较四株菌株的乙醇转化率,筛选出乙醇转化率最高的菌种进行后续试验。第三部分为发酵培养基的优化,用正交试验的方法对培养基中的氮源和无机盐进行优化。第四部分为发酵条件的优化,因素包括发酵温度、发酵初始pH和接种量。第五部分为能源甜菜的筛选与经济指标及鉴定技术的确定试验,对不同品种的甜菜进行乙醇转化率的研究,得到乙醇转化率最高的能源型甜菜。第六部分为10L发酵罐试验。
首先,在田间试验的基础上,比较四株菌发酵生产乙醇的转化率,通过发酵试验得到酿酒酵母1373的乙醇转化率最高,为72.16%,因此选择酿酒酵母1373为出发菌株。
其次,通过四因素三水平的正交试验的方法对培养基的配方进行优化,确定了最佳培养基配方为:硫酸铵0.5%、磷酸二氢钾1.0%、硫酸镁0.5%、氯化钙0.5%。在优化后的培养基配方基础上,通过单因素试验确定的发酵条件为32℃、初始pH5.5、接种量10%。
用上述条件进行不同品种甜菜的乙醇转化率的研究,筛选出能源型甜菜推广品种TYEB03、TYEB06和中甜207/09。确定能源型甜菜在酿酒酵母1373的作用下,在32℃、初始pH5.5、接种量10%的发酵条件下,发酵培养基各营养盐的添加量分别为硫酸铵0.5%、磷酸二氢钾1.0%、硫酸镁0.5%、氯化钙0.5%,在此条件下进行甜菜品种的发酵试验,根据得到的乙醇转化率的高低作为能源型甜菜一项经济技术鉴定指标。
最后,在优化的发酵培养基配方和发酵条件基础上进行了10L发酵罐的发酵试验,试验结果证实发酵罐的发酵结果要优于实验室发酵。
The energy problem is threatening the economic development of countries and affecting the living of human beings nowadays. In order to solve the energy problem, all countries in the world have carried out research on fuel ethanol. The production of fuel ethanol by microbiological fermentation becomes a focus topic in the field of research and exploitation of fuel ethanol around the world. Energy beet, which has wide planting area, simple fermentation process and high translation rate of alcohol, is a promising raw material used to produce fuel ethanol. In recent years, studies of energy beet have sprung up, however, no comprehensive identification technology of economic parameters about energy beet. In this study, different beet lines were screened by optimizing the fermentation condition of saccharomyces cerevisiae strain 1373. And the identification technology of economic parameters about energy beet was concluded.
Six parts were searched in this paper. The first part was the field experiments. The second part was the screening of the most suitable strains. Translation rates of ethanol of four strains were compared, and then the strain that had the highest ratio of ethanol was screened for the further experiment. In the third part, the medium of fermentation were optimized. The optimum content of nitrogen and different inorganic salt of ethanol fermentation was obtained from the four factor and three levels of orthogonal fitting. The fourth part was mainly about the optimization of fermentation condition, including fermentation temperature, fermentation initial pH and inoculation amount. In the fifth part, the energy beets were screened, and the economic parameters and the identification techniques were identified. The translation rates of ethanol from different beet lines were studied, in order to identify the energy beet lines which produced the highest translation rates of ethanol. In the sixth part, 10L fermentation test was made.
Firstly, based on the field experiment, the translation rates of ethanol of four strains were compared, the saccharomyces cerevisiae strain 1373 was screened by fermentation test, whose translation rate of ethanol is 72.16%. So the saccharomyces cerevisiae strain 1373 was used to do the following tests.
Secondly, the optimum composition of the medium was: NH_4SO_40.5%, CaCl_2 0.5%, KH_2PO_41.0%, MgSO_40.5%. The optimum fermentation conditions were: fermentation temperature 32℃, fermentation initial pH 5.5 and inoculation amount 10%.
The translation rates of ethanol from different beet lines were studied using the optimized fermentation conditions above. The identification technology was concluded: saccharomyces cerevisiae strain 1373 under the condition of fermentation temperature 32℃, fermentation initial pH 5.5 and inoculation amount 10%. The content of nutrients in fermentation medium was NH_4SO_40.5%, CaCl_20.5%, KH_2PO_41.0%, MgSO_40.5%. Based on the above condition the fermentation test was made. Ethanol translation rate was used as a criterion to distinguish energy beets. Three commercial lines of energy beets were screened: TYEB03, TYEB06 and zhongtian207/09.
Finally, based on the optimized fermentation medium and conditions, fermentation experiment was carried out in 10L bioreactor. The results of fermentation test in bioreactor are superior to the results in laboratory.
本试验分为六个部分。第一部分为田间试验。第二部分为最适菌株的选择试验,主要比较四株菌株的乙醇转化率,筛选出乙醇转化率最高的菌种进行后续试验。第三部分为发酵培养基的优化,用正交试验的方法对培养基中的氮源和无机盐进行优化。第四部分为发酵条件的优化,因素包括发酵温度、发酵初始pH和接种量。第五部分为能源甜菜的筛选与经济指标及鉴定技术的确定试验,对不同品种的甜菜进行乙醇转化率的研究,得到乙醇转化率最高的能源型甜菜。第六部分为10L发酵罐试验。
首先,在田间试验的基础上,比较四株菌发酵生产乙醇的转化率,通过发酵试验得到酿酒酵母1373的乙醇转化率最高,为72.16%,因此选择酿酒酵母1373为出发菌株。
其次,通过四因素三水平的正交试验的方法对培养基的配方进行优化,确定了最佳培养基配方为:硫酸铵0.5%、磷酸二氢钾1.0%、硫酸镁0.5%、氯化钙0.5%。在优化后的培养基配方基础上,通过单因素试验确定的发酵条件为32℃、初始pH5.5、接种量10%。
用上述条件进行不同品种甜菜的乙醇转化率的研究,筛选出能源型甜菜推广品种TYEB03、TYEB06和中甜207/09。确定能源型甜菜在酿酒酵母1373的作用下,在32℃、初始pH5.5、接种量10%的发酵条件下,发酵培养基各营养盐的添加量分别为硫酸铵0.5%、磷酸二氢钾1.0%、硫酸镁0.5%、氯化钙0.5%,在此条件下进行甜菜品种的发酵试验,根据得到的乙醇转化率的高低作为能源型甜菜一项经济技术鉴定指标。
最后,在优化的发酵培养基配方和发酵条件基础上进行了10L发酵罐的发酵试验,试验结果证实发酵罐的发酵结果要优于实验室发酵。
The energy problem is threatening the economic development of countries and affecting the living of human beings nowadays. In order to solve the energy problem, all countries in the world have carried out research on fuel ethanol. The production of fuel ethanol by microbiological fermentation becomes a focus topic in the field of research and exploitation of fuel ethanol around the world. Energy beet, which has wide planting area, simple fermentation process and high translation rate of alcohol, is a promising raw material used to produce fuel ethanol. In recent years, studies of energy beet have sprung up, however, no comprehensive identification technology of economic parameters about energy beet. In this study, different beet lines were screened by optimizing the fermentation condition of saccharomyces cerevisiae strain 1373. And the identification technology of economic parameters about energy beet was concluded.
Six parts were searched in this paper. The first part was the field experiments. The second part was the screening of the most suitable strains. Translation rates of ethanol of four strains were compared, and then the strain that had the highest ratio of ethanol was screened for the further experiment. In the third part, the medium of fermentation were optimized. The optimum content of nitrogen and different inorganic salt of ethanol fermentation was obtained from the four factor and three levels of orthogonal fitting. The fourth part was mainly about the optimization of fermentation condition, including fermentation temperature, fermentation initial pH and inoculation amount. In the fifth part, the energy beets were screened, and the economic parameters and the identification techniques were identified. The translation rates of ethanol from different beet lines were studied, in order to identify the energy beet lines which produced the highest translation rates of ethanol. In the sixth part, 10L fermentation test was made.
Firstly, based on the field experiment, the translation rates of ethanol of four strains were compared, the saccharomyces cerevisiae strain 1373 was screened by fermentation test, whose translation rate of ethanol is 72.16%. So the saccharomyces cerevisiae strain 1373 was used to do the following tests.
Secondly, the optimum composition of the medium was: NH_4SO_40.5%, CaCl_2 0.5%, KH_2PO_41.0%, MgSO_40.5%. The optimum fermentation conditions were: fermentation temperature 32℃, fermentation initial pH 5.5 and inoculation amount 10%.
The translation rates of ethanol from different beet lines were studied using the optimized fermentation conditions above. The identification technology was concluded: saccharomyces cerevisiae strain 1373 under the condition of fermentation temperature 32℃, fermentation initial pH 5.5 and inoculation amount 10%. The content of nutrients in fermentation medium was NH_4SO_40.5%, CaCl_20.5%, KH_2PO_41.0%, MgSO_40.5%. Based on the above condition the fermentation test was made. Ethanol translation rate was used as a criterion to distinguish energy beets. Three commercial lines of energy beets were screened: TYEB03, TYEB06 and zhongtian207/09.
Finally, based on the optimized fermentation medium and conditions, fermentation experiment was carried out in 10L bioreactor. The results of fermentation test in bioreactor are superior to the results in laboratory.
引文
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