高效油脂降解菌株的选育及其降解条件的初步研究
摘要
餐饮废水是一种高浓度的油脂废水，如不经处理直接排放水体，影响水体的复氧及其自净化过程，危害水体生态系统。目前处理这种废水的物理化学方法都不尽理想，无法彻底消除废水中的油脂，而生物学方法处理污水是最有效合理的方法、无二次污染，因此广泛受到国内外学者的重视。
本研究从学校餐厅下水道口、油脂加工厂及屠宰场的含油废水及土壤中采集土样，利用有效的初筛和复筛体系，经过大量的菌种筛选，分离得到一株降解油脂能力较高的菌株DC-3-2，其降解率最大，达到67.32％。经细胞、菌落、子囊孢子及假菌丝的形态学观察和糖发酵实验、碳源同化、硝酸盐同化、产类淀粉化合物、尿素分解、明胶液化、无维生素培养基生长等形态学和各生理生化指标的实验，并按照J．Lodder酵母分类系统，初步鉴定DC-3-2菌株为假丝酵母属(Candida sp．)。
利用离子束诱变，以筛选出的Candida sp．为出发菌株，进行能量30kev和各个剂量的N~+注入，结果显示：在4×10~(15)ion／cm~2时，正突变率最高，因此确定以4×10~(15)ion／cm~2为诱变剂量。在该剂量下，对出发菌株进行诱变。结果表明，诱变后的菌株DC-3-2-50与出发菌株DC-3-2对油脂的降解率相比，其降解率提高了11.09％。利用单因素实验，研究了pH值、接种量、温度、摇床转速及不同油脂对油脂降解率的影响。实验结果表明，最佳培养条件：初始pH值9.0、接种量3.0％、温度为25-30℃、摇床转速为180～200rpm、碳源为大豆色拉油。在优化好的条件下，其降解率最高，可达87.7％。
Restaurant wastewater is a high-concentration oil wastewater. If it is not disposed and discharged to water directly, it will have a serious influence on the process of reoxygenation and self-purification, and jeopardizes aqueous ecosystem. Current physical and chemical technologies such flotation and sedimentation are often insufficient and remove oil in water incompletely. Biological technology is the most effective safe and reasonable method for wastewater, and does not bring secondary pollution. Therefore, all the scholars at home and abroad regard them very well.
In the study, lipase-producing strain was isolated from oil-contained sewage and earth samples in cloaca of university, oil manufactory, slaughterhouse. Oil-degrading rate of a strain DC-3-2 was the highest after effectively primary and secondary screening method, which reached 67.32%. Several experiments were done: morphology observation of cell , colony, ascospore and pseudo-hyphae and so on, and physiology biochemistry, such as glucose fermentation, carbon nutriments assimilation, nitrogen nutriments assimilation, producing starch compound, urealyticum, gelatin liquefaction, growth in no-vitamin plates. According to J.Lodders yeast classification system, the yeast was identified as Candida sp. primarily.
On the base of this, the original strain Candida sp. DC-3-2 was implanted by 30kev N~+ in several doses. The results showed that the plus mutant percentage was the highest in the dose of 4×10~(15)ion/cm~2.Therefore, the dose was conformed optimum and the mutant was performed in the dose. The results showed that the oil-degrading rate of the strain implanted by N~+ ion beam was increased by 11.09%. Single factor method was used in the experiments to study the influence on oil-degrading rate. There were several factors: pH value, inoculum size, temperature, dissolved oxygen, carbon nutriments. The results indicated that the optimal culture conditions were as follow: initial pH value 9.0, inoculum size 3%, temperature 25～30°C, dissolved oxygen 180～200rpm, carbon nutriments soybean salad oil. In optimum condition, it had the highest oil-degrading rate, which was 87.7%.
本研究从学校餐厅下水道口、油脂加工厂及屠宰场的含油废水及土壤中采集土样，利用有效的初筛和复筛体系，经过大量的菌种筛选，分离得到一株降解油脂能力较高的菌株DC-3-2，其降解率最大，达到67.32％。经细胞、菌落、子囊孢子及假菌丝的形态学观察和糖发酵实验、碳源同化、硝酸盐同化、产类淀粉化合物、尿素分解、明胶液化、无维生素培养基生长等形态学和各生理生化指标的实验，并按照J．Lodder酵母分类系统，初步鉴定DC-3-2菌株为假丝酵母属(Candida sp．)。
利用离子束诱变，以筛选出的Candida sp．为出发菌株，进行能量30kev和各个剂量的N~+注入，结果显示：在4×10~(15)ion／cm~2时，正突变率最高，因此确定以4×10~(15)ion／cm~2为诱变剂量。在该剂量下，对出发菌株进行诱变。结果表明，诱变后的菌株DC-3-2-50与出发菌株DC-3-2对油脂的降解率相比，其降解率提高了11.09％。利用单因素实验，研究了pH值、接种量、温度、摇床转速及不同油脂对油脂降解率的影响。实验结果表明，最佳培养条件：初始pH值9.0、接种量3.0％、温度为25-30℃、摇床转速为180～200rpm、碳源为大豆色拉油。在优化好的条件下，其降解率最高，可达87.7％。
Restaurant wastewater is a high-concentration oil wastewater. If it is not disposed and discharged to water directly, it will have a serious influence on the process of reoxygenation and self-purification, and jeopardizes aqueous ecosystem. Current physical and chemical technologies such flotation and sedimentation are often insufficient and remove oil in water incompletely. Biological technology is the most effective safe and reasonable method for wastewater, and does not bring secondary pollution. Therefore, all the scholars at home and abroad regard them very well.
In the study, lipase-producing strain was isolated from oil-contained sewage and earth samples in cloaca of university, oil manufactory, slaughterhouse. Oil-degrading rate of a strain DC-3-2 was the highest after effectively primary and secondary screening method, which reached 67.32%. Several experiments were done: morphology observation of cell , colony, ascospore and pseudo-hyphae and so on, and physiology biochemistry, such as glucose fermentation, carbon nutriments assimilation, nitrogen nutriments assimilation, producing starch compound, urealyticum, gelatin liquefaction, growth in no-vitamin plates. According to J.Lodders yeast classification system, the yeast was identified as Candida sp. primarily.
On the base of this, the original strain Candida sp. DC-3-2 was implanted by 30kev N~+ in several doses. The results showed that the plus mutant percentage was the highest in the dose of 4×10~(15)ion/cm~2.Therefore, the dose was conformed optimum and the mutant was performed in the dose. The results showed that the oil-degrading rate of the strain implanted by N~+ ion beam was increased by 11.09%. Single factor method was used in the experiments to study the influence on oil-degrading rate. There were several factors: pH value, inoculum size, temperature, dissolved oxygen, carbon nutriments. The results indicated that the optimal culture conditions were as follow: initial pH value 9.0, inoculum size 3%, temperature 25～30°C, dissolved oxygen 180～200rpm, carbon nutriments soybean salad oil. In optimum condition, it had the highest oil-degrading rate, which was 87.7%.
引文
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