【作者】 王艳；

【导师】 辛嘉英；

【作者基本信息】 哈尔滨商业大学 ， 发酵工程， 2010， 硕士

【摘要】 开展生物降解萘的研究,不仅可以消除环境中残存的萘达到治理环境污染的目的,而且可以实现萘的生物转化和生物降解,对社会的发展和绿色化工产业的建立都有重要的意义。萘是最常见的一种多环芳烃,目前世界上许多水体，包括湖泊甚至地下水中都有萘的存在,鉴于其潜在的毒害作用,其在自然环境中越来越高的浓度已引起人们的密切关注。靛蓝广泛应用于印染、医药和食品工业,然而目前的化学合成靛蓝反应中大量使用的苯胺是一种潜在的致癌物质。因此,采用生物转化法生产靛蓝,在降低生产成本、减少能耗、生产安全及环境安全等方面无疑都具有重要意义。为了提高帕氏氢噬胞菌(Hydrogenophaga Palleronii) LHJ38在以茶为唯一碳源和能源的培养基中的生长能力和萘的降解能力,本实验将两相体系技术合理的应用于其中。分别测定了该菌株在单相体系和两相体系中的生长曲线和萘降解曲线,同时对经两相体系培养后的油相细胞和水相细胞的生长能力和降解能力进行了深入研究。研究结果表明：最佳的两相体系是由石蜡油和金属盐培养基(3：100)组成的反应系统,其中萘的最佳浓度为2g/L。Hydrogenophaga Palleronii LHJ38在此两相体系中培养72h后最大生物量达到4.248g/L是单相反应系统的4倍。而且该两相体系中反应96h后92%的萘被降解掉,132h内浓度为2g/L萘完全被降解,萘的降解能力比在单相系统中的降解能力高出两倍还多。经两相体系培养后的细胞萘降解能力也大大提高,而且从油相中获得的细胞的萘降解速率比从水相中获得的细胞要快2.7倍。萘降解菌株Hydrogenophaga Palleronii LHJ38在液体培养摹和固体琼脂培养基上能够利用吲哚生物合成靛蓝。在LB培养基中不加其他诱导剂就能迅速合成靛蓝,以LB培养基为能源和碳源的Hydrogenophaga Palleronii LHJ38生物合成靛蓝的最大数率为25.3mg min’g(dry weight)’。Hydrogenophaga Palloronii LHJ38在金属盐培养基中有微弱的生物合成能力,但是在金属盐培养基中加入水杨酸钠,经过两天的诱导培养,Hydrogenophaga Palleronii LHJ38整细胞生物合成靛蓝的能力比没有经过水杨酸钠诱导大为提高,其生物合成靛蓝的最大浓度达9.99mg/L。经过水杨酸钠诱导的Hydrogenophaga Palleronii LHJ38整细胞生物合成靛蓝的能力提高了77倍。Hydrogenophaga Palleronii LHJ38在LB培养基中培养两天,在缓冲溶液中合成靛蓝的最高浓度为19.71mg/L,其生物合成靛蓝的能力为加入水杨酸钠时的2.80倍。与其最适生长pH值不同,Hydrogenophaga Palleronii LHJ38生物合成靛蓝的最佳pH范围是8.45-9.45,在pH为8.95的情况下生物合成靛蓝的能力是其在pH值为6.95的情况下的两倍。更多还原

【Abstract】 The biodegradation of naphthalene can eliminate naphthalene in environment to decrease pollution. In addition, as catalyst, enzyme or cells in biodegradation process which may involve in biotransformation of naphthalene, have important significance to social development and green chemical industry. Naphthalene is the most familiar polycyclic aromatic hydrocarbon and exists in all of the waters in the world. Due to the potential toxicity, and the higher and higher concentration in environment, naphthalene has evoked abroad attention in the world range. Indigo is a traditional pigment extensively used in the printing and dyeing, pharmaceutical and food industry. Aniline is a latent carcinogenic substance which is vastly used in the reaction of chemosynthesis of indigo. Biosynthesis of indigo has signality in reduce operating costs and decrease energy consumption and enhance the manufacture security and environment security.Naphthalene-degradaing bacteria Hydrogenophaga Pallcronii LHJ38 can grow on naphthalene as sole energy and carbon source. The growth and naphthalene degradation curve of the strain were determined in biphase and monophase system respectively. The growth and naphthalene degradation activity of cells from oil phase and aqueous phase were compared carefully. It is higher more than 4 fold, the maximum biomass of 4.248g/l. was achieved after 72h incubation under the optimum conditions of 3mL paraffin oil and 0.2g naphthalene in 100mL mineral salt medium (MSM), than the MSM monophase system. In the biphasc system, the naphthalene degradation capacity of Hydrogenophaga Palleronii LHJ38 was higher more than 2 times than that in the monophase system. The naphthalene (2g/l.) can be degraded completely in 132h by adding 10mL·cell suspension (0.2mg dry weight cell/mL). The naphthalene degradation rate of cells from oil phase was 2.7 fold as fast as that of cells from aqueous phase.The biosynthesis of indigo by naphthalene-degradaing bacteria Hydrogenophaga Palleronii LHJ38 from indole in mineral medium and on solid agar medium has been studied. Indigo formation capacity by Hydrogenophaga Palleronii LHJ38 cell growing in mineral salts medium in the presence of salicylate greatly increased. Contrarily. indigo formation capacity by Hydrogenophaga Palleronii LHJ38 cell growing in LB medium in the presence of salicylate decreased by 2 fold. The optimum pH ranges for biosnythesis of indigo formation by Hydrogenophaga Palleronii LHJ38 growing in LB medium at pH 8.95 increased 2 fold compared with that at pH 6.95.更多还原