马尾藻发酵生产燃料乙醇的研究
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摘要
在生物质能源中,乙醇作为替代性再生能源之一,具有燃烧安全、效率高、无污染等特点,燃料乙醇的研究开发已成为一项世界重大的热门课题。然而伴随着人口的增加,可利用的耕地不断的减少,粮食和土地的供给成为燃料乙醇发展的瓶颈。海藻与陆地植物相比,具有繁殖速度快、产量高、不占用耕地等特点。褐藻等海藻类植物中含有大量的碳水化合物,而这些碳水化合物能够被细菌、酵母菌等微生物直接或间接发酵转化为燃料乙醇。
本文以广两省北海市涠洲岛海域内的马尾藻作为原料,在实验室条件下,首先通过对原料马尾藻的化学组成进行分析；然后以马尾藻中主要成分的甘露崞作为研究对象,利用Zymobacter palmae菌株通过正交实验分析法进行生产燃料乙醇的条件优化研究；最后在适当条件下利用硫酸溶液处理马尾藻制备提取液,再接入工业酵母菌进行水解液发酵生产乙醇。
实验结果表明,马尾藻中组成成分主要由5.626%的粗脂肪、11.585%粗蛋白、29.183%灰分及53.606%碳水化合物构成,特别是碳水化合物中还原糖的含量达到29.4%。发酵过程中主要是将部分碳水化合物转化为乙醇。
Zymobacter palmae在250m1三角烧瓶中发酵甘露醇生产乙醇的最适培养基为：甘露醇20.0g/L,酵母膏0.5g/L; MgSO4·7H2O0.1g/L;KH2PO42.0g/L; K2HPO47.0g/L;(NH4)2SO41.0g/L,PH=6.0。通过正交分析实验得到Z. palmae利用甘露醇发酵生产乙醇的最佳条件为摇床转速150rpm,温度28。C,发酵液体积150ml。在最佳发酵条件下,乙醇的最大产量为0.429g乙醇儋底物,乙醇转化率达到理论值的84%。
在氮源及其他营养成分固定的条件下,工业酵母菌R1-11利用马尾藻提取液发酵生产乙醇的含量为0.385%(V/V),而工业酵母菌R1-12生产乙醇的含量为0.275%(V/V),对比可以得出,R1-11利用马尾藻提取液生产乙醇的能力要高于工业酵母R1-12。R1-11在含有外加氮源的培养基中生产得到的乙醇含量为0.594%(V/V),R1-11在无外加氮源的发酵培养基中发酵生产的得到的乙醇含量为0.385%(V/V)。因此,碳氮比的比例对于利用马尾藻提取液提高生产乙醇含量有重要影响。
As one of biomass energy sources, ethanol has been considered to be an excellent alternative renewable fuel with characteristics in the safety, efficiency and non-pollution of combustion. Researches and developments of fuel ethanol has become a worldwide topical issue. However, along with the increase of population and the reduction of arable land, the supply of food and land become the bottleneck in the development of fuel ethanol. By comparison with terrestrial plants, algae grows faster, its yield is higher and it avoids the use of land. Algae such as brown seaweed contains rich carbohydrates which can be used to produce fuel alcohol directly or indirectly through bioconversion fermented with bacteria, yeast and other microorganisms.
In this paper, Sargasso from the sea area of Weizhou Island, Beihai, Guangxi, is used as raw materials to analyze chemical composition, then mannitol which is considered to be the main component in Sargasso was used as research objects to investigate condition optimization of ethanol fermentation with Zymobacter palmae strain. Last, sulfuric acid was used to degrade the Sargasso into extract which used for ethanol fermentation by industrial yeast. The experimental results showed that the main compositions of Sargasso was combined with5.626%crude fat,11.585%crude protein,29.138%ash and53.606%carbohydrates with29.4%of reducing sugar in the carbohydrates. The part of carbohydrates was used to carry out ethanol fermentation; The optimal medium which mannitol was inverted to ethanol at high efficiency by Zymobacter palmae in250ml flask was mannitol20.Og/L, yeast extract0.5g/L, MgSO4·7H2O0.1g/L, KH2PO42.0g/L, K2HPO47.0g/L,(NH4)2SO41.0g/L, PH=6.0; And the best fermentation conditions to produce ethanol was the rotation speed of150rpm, temperature of28℃and fermentation liquor of150ml with a yield of0.429g ethanol/g mannitol. Ethanol conversion rate achieved the theoretical value of84%; Under conditions of fixed nitrogen and other nutrients, industrial yeast, R1-11, was use to produce ethanol in Sargasso extract, and the yield of ethanol was0.385%(V/V), but the ethanol yield of another industrial yeast, R1-12, was only0.275%(V/V).The ability of ethanol fermentation with Sargasso extract by R1-11was obviously higher than R1-12. The production of ethanol fermentation by R1-11was0.594%(V/V) in the medium with nitrogen addition, and the production of ethanol fermentation by R1-11was0.385%(V/V) in the medium without nitrogen addition. Therefore, the ratio of carbon and nitrogen has an important impaction on the Sargasso extract to improve the production of ethanol content.
本文以广两省北海市涠洲岛海域内的马尾藻作为原料,在实验室条件下,首先通过对原料马尾藻的化学组成进行分析；然后以马尾藻中主要成分的甘露崞作为研究对象,利用Zymobacter palmae菌株通过正交实验分析法进行生产燃料乙醇的条件优化研究；最后在适当条件下利用硫酸溶液处理马尾藻制备提取液,再接入工业酵母菌进行水解液发酵生产乙醇。
实验结果表明,马尾藻中组成成分主要由5.626%的粗脂肪、11.585%粗蛋白、29.183%灰分及53.606%碳水化合物构成,特别是碳水化合物中还原糖的含量达到29.4%。发酵过程中主要是将部分碳水化合物转化为乙醇。
Zymobacter palmae在250m1三角烧瓶中发酵甘露醇生产乙醇的最适培养基为：甘露醇20.0g/L,酵母膏0.5g/L; MgSO4·7H2O0.1g/L;KH2PO42.0g/L; K2HPO47.0g/L;(NH4)2SO41.0g/L,PH=6.0。通过正交分析实验得到Z. palmae利用甘露醇发酵生产乙醇的最佳条件为摇床转速150rpm,温度28。C,发酵液体积150ml。在最佳发酵条件下,乙醇的最大产量为0.429g乙醇儋底物,乙醇转化率达到理论值的84%。
在氮源及其他营养成分固定的条件下,工业酵母菌R1-11利用马尾藻提取液发酵生产乙醇的含量为0.385%(V/V),而工业酵母菌R1-12生产乙醇的含量为0.275%(V/V),对比可以得出,R1-11利用马尾藻提取液生产乙醇的能力要高于工业酵母R1-12。R1-11在含有外加氮源的培养基中生产得到的乙醇含量为0.594%(V/V),R1-11在无外加氮源的发酵培养基中发酵生产的得到的乙醇含量为0.385%(V/V)。因此,碳氮比的比例对于利用马尾藻提取液提高生产乙醇含量有重要影响。
As one of biomass energy sources, ethanol has been considered to be an excellent alternative renewable fuel with characteristics in the safety, efficiency and non-pollution of combustion. Researches and developments of fuel ethanol has become a worldwide topical issue. However, along with the increase of population and the reduction of arable land, the supply of food and land become the bottleneck in the development of fuel ethanol. By comparison with terrestrial plants, algae grows faster, its yield is higher and it avoids the use of land. Algae such as brown seaweed contains rich carbohydrates which can be used to produce fuel alcohol directly or indirectly through bioconversion fermented with bacteria, yeast and other microorganisms.
In this paper, Sargasso from the sea area of Weizhou Island, Beihai, Guangxi, is used as raw materials to analyze chemical composition, then mannitol which is considered to be the main component in Sargasso was used as research objects to investigate condition optimization of ethanol fermentation with Zymobacter palmae strain. Last, sulfuric acid was used to degrade the Sargasso into extract which used for ethanol fermentation by industrial yeast. The experimental results showed that the main compositions of Sargasso was combined with5.626%crude fat,11.585%crude protein,29.138%ash and53.606%carbohydrates with29.4%of reducing sugar in the carbohydrates. The part of carbohydrates was used to carry out ethanol fermentation; The optimal medium which mannitol was inverted to ethanol at high efficiency by Zymobacter palmae in250ml flask was mannitol20.Og/L, yeast extract0.5g/L, MgSO4·7H2O0.1g/L, KH2PO42.0g/L, K2HPO47.0g/L,(NH4)2SO41.0g/L, PH=6.0; And the best fermentation conditions to produce ethanol was the rotation speed of150rpm, temperature of28℃and fermentation liquor of150ml with a yield of0.429g ethanol/g mannitol. Ethanol conversion rate achieved the theoretical value of84%; Under conditions of fixed nitrogen and other nutrients, industrial yeast, R1-11, was use to produce ethanol in Sargasso extract, and the yield of ethanol was0.385%(V/V), but the ethanol yield of another industrial yeast, R1-12, was only0.275%(V/V).The ability of ethanol fermentation with Sargasso extract by R1-11was obviously higher than R1-12. The production of ethanol fermentation by R1-11was0.594%(V/V) in the medium with nitrogen addition, and the production of ethanol fermentation by R1-11was0.385%(V/V) in the medium without nitrogen addition. Therefore, the ratio of carbon and nitrogen has an important impaction on the Sargasso extract to improve the production of ethanol content.
引文
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