气升式反应器中模拟燃料油的生物脱硫特性研究
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摘要
石油是当今世界的主要能源之一,全球每天约消耗1000多万吨的原油,这些燃料油燃烧后产生的SO_2是形成酸雾、酸雨等环境污染的重要原因。低含硫矿物燃料的稀缺、高品质清洁燃料油的需求和日益严格的控制标准迫切需要开发燃烧前深度脱硫技术。加氢技术是目前的主要脱硫法,然而其在操作成本和深度脱硫方面的不足制约着脱硫技术的发展。生物脱硫具有反应条件温和、专一性强、不影响油品热值等优点,已成为加氢脱硫工艺较好的补充,随着世界范围内对油品含硫标准的日益严格,作为深度脱硫技术的生物脱硫受到越来越多的关注。
本研究利用实验室自主分离的脱硫菌株分支杆菌ZD-19,以促进生物脱硫的工业应用为目标进行研究。课题前期研究证明ZD-19可以沿4S途径降解二苯并噻吩(DBT)和4,6二甲基二苯并噻吩(4,6DMDBT),并且具有较高的去除效率。为了进一步考察ZD-19的脱硫底物范围,本论文就ZD-19对4甲基二苯并噻吩(4MDBT)的脱硫特性进行了研究,结果表明ZD-19也能专一性的切断4MDBT的C-S键,且由于4MDBT独特的不对称结构,代谢过程中羟基取代位置发生变化,所得产物为两个同分异构体:4甲基2苯基苯酚和2羟基3甲基联苯。研究还证实ZD-19并不能有效降解苯并噻吩(BT)。本部分实验表明ZD-19是一株对DBT及其衍生物有较高去除效率的优势菌种。
微生物能够在油相中作用是BDS应用于实际油品脱硫的关键。摇瓶实验中,ZD-19在油水两相体系中具有较高的脱硫活性,反应10h,1mmol/LDBT的降解效率达到78.5%;其最佳脱硫活性是水相中的2倍,且两相体系促进了传质,提高了细胞的利用效率,较高菌液浓度时没有出现抑制现象,具有比水相实验较高的脱硫活性。与摇床实验比较,气升式反应器独特的流体力学特征更加利于两相混合和传质,在生长细胞脱硫和休止细胞脱硫实验中都表现出明显的优势。反应器内ZD-19达到指数生长后期时细胞最大比生长速率和细胞浓度为0.15g/(L·h)和8.1g/L,分别是摇床实验1.4倍和2.3倍。ZD-19在油水两相实验中表现出较高的脱硫活性,具有应用于油品脱硫的潜力。
目前BDS研究还局限于摇瓶实验,生物脱硫反应器的设计与研究具有重要意义。本研究在自行设计的气升式反应器中生物降解正十六烷中的DBT,对气升式反应器的基本工艺参数进行优化。在实验所选的条件下,最佳表观速率为11.8cm/s。表观速率达到13.0cm/s时,出现轻微柱塞流现象,液滴直径增大,传质效果开始降低。反应器中ZD-19休止细胞在生长介质(培养基)和非生长介质(磷酸缓冲溶液)中具有几乎相同的脱硫活性;过低的菌液浓度下不仅两相界面小,传质速率低,也使DBT对细胞的毒性作用明显,脱硫效率很差,实验条件下10-40g/L的菌液浓度都具有较好的比脱硫速率;增加DBT初始浓度,脱硫量略有增加,直至高DBT的致毒作用使菌液脱硫活性降低,此时传质速率较高,体系属于反应控制。
在气升式反应器内,立足于考察ZD-19在模拟燃料油体系中的脱硫特性,实验以柴油中含量较多的有机硫化合物DBT、4MDBT和4,6DMDBT为模型化合物,设计了单一底物和混合底物实验。单底物脱硫实验表明,ZD-19对DBT、4MDBT和4,6DMDBT的脱硫能力为DBT≈4MDBT＞4,6DMDBT。4MDBT的高脱硫效率和对细胞的低毒性,使得混合体系下的总硫去除率更高;双底物脱硫实验和混合脱硫实验共同表明,ZD-19对多种有机硫共存时的脱硫效果良好,具有应用于实际油品脱硫的潜力,有很广的应用前景。
About ten million tons of crude oil was consumed every day, resulting in the emission of sulfur-oxides to the atmosphere, which is a main cause of serious environmental problems such as acid rain. High sulfur contents of the crude oil reserves, the need for fuels ultra-low in sulfur and continued stringent regulations call for deep desulfurization technologies. Hydrodesulfurization (HDS) is the dominating method to desulfurize oil, but its defect of high operation cost and deep desulfurization limited the development of desulfurization technology. Biodesulfurization (BDS) which can selectively desulfurize the most recalcitrant sulfur compounds from oil under mild conditions, remaining the combustion value of the fuels, support HDS very well. As stricter standard of sulfur content in oil appears, BDS gains more attention.
Mycobacterium sp. ZD-19 can metabolize dibenzothiophene(DBT) and 4,6dimethyldibenzothiophene(4,6DMDBT) through a sulfur-specific pathway. The biodesulfurization of 4methyldibezothiophene(4MDBT) by ZD-19 was also investigated in this thesis. The results showed that ZD-19 can also desulfurize 4MDBT through a sulfur-specific pathway. The metabolic pathway for biodesulfurization of 4MDBT was studied, and the final product of 4MDBT biodesulfurization was determined by GC-MS, which were identified as two isomers: 4-methyl-2phenylphenol and 2-hydroxy-3-methyl-biphenyl.
Secondly, biodesulfurization by resting cells of ZD-19 was investigated of biphasic system in shake flasks scale. The results indicated that, ZD-19 in oil-aqueous system had twice the desulfurization activity of aqueous phase, because inhibitions effects by product accumulation in aqueous media were not so clear in biphasic condition as 2-hydroxybiphenyl (HBP) oil-water partition coefficient is very high; and mass transfer limitation was partially eliminated. In contrast to shake flasks, biodesulfurization in a self-designed airlift bioreactor(ALR) can achieve higher desulfurization rate; the maximum specific growth rate and the highest cell concentration at the late exponential growth phase in the reactor were 0.15g/(L·h) and 8.1g/L, which were 1.4 and 2.3 times that of shake flasks respectively. The results showed ALR's great potential for the commercial use of biodesulfurization to remove sulfur from fuel oil.
Biodesulfurization of DBT in oil-aqueous system was carried out in the ALR. The optimal gas velocity was 11.8cm/s. Slight plug-flow was formed when the gas velocity achieved 13.0cm/s, and the oil droplet diameter and the interface area decreased consequently, which were the main causes of the decreased desulfurization rate. The biodesulfurization of resting cells of ZD-19 could be proceed in the non-growth media (phosphate buffer) as efficiently as in the growth medium (BSM). The desulfurization rate increased with the increasing of DBT concentration as well, while high DBT concentration would inhibit the activity with an optimal of 3 mmol·L~(-1). The results suggested that under high concentration of DBT and optimal gas velocity, the mass transfer rate was much higher compared to DBT conversion rates, when the microbial desulfurization rate was the overall rate-limiting process step.
Lastly, The desulfurization ability of Mycobacterium sp. ZD-19 for DBT and alkylated dibenzothiophenes (Cx-DBTs) was studied of stimulant fuel oil system in the ALR. The desulfurization of DBT, 4MDBT or 4,6-dimethyldibezothiophene ( 4, 6-DMDBT) as a sole substrate, and the mixture of two or three of them as sulfur substrate were investigated respectively. The results suggested that this strain could efficiently desulfurize DBT and Cx-DBTs, their activity was: DBT≈4MDBT>4,6DMDBT. The degradation rate of total sulfur related to the composition and structure of sulfur substrates. The total sulfur remove percentage when DBT and 4MDBT were mixed was larger than any other cases, which attributed to the high desulfurization rate of 4MDBT and its little negative effect on cells activities.
Biodesulfurization of DBT derivatives such as 4-MDBT and 4,6-DMDBT demonstrated Mycobacterium sp. ZD-19's ability to attack a wide range of sulfur compounds present in diesel oil. The activity of the strain shows its possibility of the commercial use for decreasing the sulfur content of diesel fuels.
本研究利用实验室自主分离的脱硫菌株分支杆菌ZD-19,以促进生物脱硫的工业应用为目标进行研究。课题前期研究证明ZD-19可以沿4S途径降解二苯并噻吩(DBT)和4,6二甲基二苯并噻吩(4,6DMDBT),并且具有较高的去除效率。为了进一步考察ZD-19的脱硫底物范围,本论文就ZD-19对4甲基二苯并噻吩(4MDBT)的脱硫特性进行了研究,结果表明ZD-19也能专一性的切断4MDBT的C-S键,且由于4MDBT独特的不对称结构,代谢过程中羟基取代位置发生变化,所得产物为两个同分异构体:4甲基2苯基苯酚和2羟基3甲基联苯。研究还证实ZD-19并不能有效降解苯并噻吩(BT)。本部分实验表明ZD-19是一株对DBT及其衍生物有较高去除效率的优势菌种。
微生物能够在油相中作用是BDS应用于实际油品脱硫的关键。摇瓶实验中,ZD-19在油水两相体系中具有较高的脱硫活性,反应10h,1mmol/LDBT的降解效率达到78.5%;其最佳脱硫活性是水相中的2倍,且两相体系促进了传质,提高了细胞的利用效率,较高菌液浓度时没有出现抑制现象,具有比水相实验较高的脱硫活性。与摇床实验比较,气升式反应器独特的流体力学特征更加利于两相混合和传质,在生长细胞脱硫和休止细胞脱硫实验中都表现出明显的优势。反应器内ZD-19达到指数生长后期时细胞最大比生长速率和细胞浓度为0.15g/(L·h)和8.1g/L,分别是摇床实验1.4倍和2.3倍。ZD-19在油水两相实验中表现出较高的脱硫活性,具有应用于油品脱硫的潜力。
目前BDS研究还局限于摇瓶实验,生物脱硫反应器的设计与研究具有重要意义。本研究在自行设计的气升式反应器中生物降解正十六烷中的DBT,对气升式反应器的基本工艺参数进行优化。在实验所选的条件下,最佳表观速率为11.8cm/s。表观速率达到13.0cm/s时,出现轻微柱塞流现象,液滴直径增大,传质效果开始降低。反应器中ZD-19休止细胞在生长介质(培养基)和非生长介质(磷酸缓冲溶液)中具有几乎相同的脱硫活性;过低的菌液浓度下不仅两相界面小,传质速率低,也使DBT对细胞的毒性作用明显,脱硫效率很差,实验条件下10-40g/L的菌液浓度都具有较好的比脱硫速率;增加DBT初始浓度,脱硫量略有增加,直至高DBT的致毒作用使菌液脱硫活性降低,此时传质速率较高,体系属于反应控制。
在气升式反应器内,立足于考察ZD-19在模拟燃料油体系中的脱硫特性,实验以柴油中含量较多的有机硫化合物DBT、4MDBT和4,6DMDBT为模型化合物,设计了单一底物和混合底物实验。单底物脱硫实验表明,ZD-19对DBT、4MDBT和4,6DMDBT的脱硫能力为DBT≈4MDBT＞4,6DMDBT。4MDBT的高脱硫效率和对细胞的低毒性,使得混合体系下的总硫去除率更高;双底物脱硫实验和混合脱硫实验共同表明,ZD-19对多种有机硫共存时的脱硫效果良好,具有应用于实际油品脱硫的潜力,有很广的应用前景。
About ten million tons of crude oil was consumed every day, resulting in the emission of sulfur-oxides to the atmosphere, which is a main cause of serious environmental problems such as acid rain. High sulfur contents of the crude oil reserves, the need for fuels ultra-low in sulfur and continued stringent regulations call for deep desulfurization technologies. Hydrodesulfurization (HDS) is the dominating method to desulfurize oil, but its defect of high operation cost and deep desulfurization limited the development of desulfurization technology. Biodesulfurization (BDS) which can selectively desulfurize the most recalcitrant sulfur compounds from oil under mild conditions, remaining the combustion value of the fuels, support HDS very well. As stricter standard of sulfur content in oil appears, BDS gains more attention.
Mycobacterium sp. ZD-19 can metabolize dibenzothiophene(DBT) and 4,6dimethyldibenzothiophene(4,6DMDBT) through a sulfur-specific pathway. The biodesulfurization of 4methyldibezothiophene(4MDBT) by ZD-19 was also investigated in this thesis. The results showed that ZD-19 can also desulfurize 4MDBT through a sulfur-specific pathway. The metabolic pathway for biodesulfurization of 4MDBT was studied, and the final product of 4MDBT biodesulfurization was determined by GC-MS, which were identified as two isomers: 4-methyl-2phenylphenol and 2-hydroxy-3-methyl-biphenyl.
Secondly, biodesulfurization by resting cells of ZD-19 was investigated of biphasic system in shake flasks scale. The results indicated that, ZD-19 in oil-aqueous system had twice the desulfurization activity of aqueous phase, because inhibitions effects by product accumulation in aqueous media were not so clear in biphasic condition as 2-hydroxybiphenyl (HBP) oil-water partition coefficient is very high; and mass transfer limitation was partially eliminated. In contrast to shake flasks, biodesulfurization in a self-designed airlift bioreactor(ALR) can achieve higher desulfurization rate; the maximum specific growth rate and the highest cell concentration at the late exponential growth phase in the reactor were 0.15g/(L·h) and 8.1g/L, which were 1.4 and 2.3 times that of shake flasks respectively. The results showed ALR's great potential for the commercial use of biodesulfurization to remove sulfur from fuel oil.
Biodesulfurization of DBT in oil-aqueous system was carried out in the ALR. The optimal gas velocity was 11.8cm/s. Slight plug-flow was formed when the gas velocity achieved 13.0cm/s, and the oil droplet diameter and the interface area decreased consequently, which were the main causes of the decreased desulfurization rate. The biodesulfurization of resting cells of ZD-19 could be proceed in the non-growth media (phosphate buffer) as efficiently as in the growth medium (BSM). The desulfurization rate increased with the increasing of DBT concentration as well, while high DBT concentration would inhibit the activity with an optimal of 3 mmol·L~(-1). The results suggested that under high concentration of DBT and optimal gas velocity, the mass transfer rate was much higher compared to DBT conversion rates, when the microbial desulfurization rate was the overall rate-limiting process step.
Lastly, The desulfurization ability of Mycobacterium sp. ZD-19 for DBT and alkylated dibenzothiophenes (Cx-DBTs) was studied of stimulant fuel oil system in the ALR. The desulfurization of DBT, 4MDBT or 4,6-dimethyldibezothiophene ( 4, 6-DMDBT) as a sole substrate, and the mixture of two or three of them as sulfur substrate were investigated respectively. The results suggested that this strain could efficiently desulfurize DBT and Cx-DBTs, their activity was: DBT≈4MDBT>4,6DMDBT. The degradation rate of total sulfur related to the composition and structure of sulfur substrates. The total sulfur remove percentage when DBT and 4MDBT were mixed was larger than any other cases, which attributed to the high desulfurization rate of 4MDBT and its little negative effect on cells activities.
Biodesulfurization of DBT derivatives such as 4-MDBT and 4,6-DMDBT demonstrated Mycobacterium sp. ZD-19's ability to attack a wide range of sulfur compounds present in diesel oil. The activity of the strain shows its possibility of the commercial use for decreasing the sulfur content of diesel fuels.
引文
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