Alkane and polycyclic aromatic hydrocarbon degrading compound bacterial agent and preparing method thereof

Abstract

The invention relates to an alkane and polycyclic aromatic hydrocarbon degrading compound bacterial agent and a preparing method thereof. An alkane-degrading strain gordonia terrae NR-61 and a polycyclic aromatic hydrocarbon-degrading strain pseudomonas stutzeri ZJ-7 are fermented respectively by a special culture medium; thalli cells are obtained by centrifugation in a clarified dish-type centrifuge; dextrin, kaolin, (NH4)2HPO4 and sodium dodecyl sulfate are added to the thalli cells in proportion successively, and an obtained white solid preparation is the target product. The alkane and polycyclic aromatic hydrocarbon degrading compound bacterial agent with high efficiency and environmental friendliness can have the degrading rate of alkane in polluted soil reaching 75% or more and the degrading rate of polycyclic aromatic hydrocarbons (naphthalene and phenanthrene) reaching 70% or more, and has the advantages of economy and environmental protection, low implementation cost, simple operation and the like.

Description

1. Technical field:

[0001] The invention relates to a compound bacterial agent for degrading alkanes and polycyclic aromatic hydrocarbons and a preparation method thereof. 2. Background technology:

[0002] Alkanes and polycyclic aromatic hydrocarbons are common toxic, harmful and refractory organic pollutants that seriously endanger the ecological environment and human health. Oil spills, incomplete combustion of fuels such as coal and petroleum, farmland sewage irrigation, and atmospheric deposition are the main sources of them. . Physicochemical methods (chemical oxidation method, thermodynamic repair method, isolation method, photocatalytic method, etc.) can treat alkanes and polycyclic aromatic hydrocarbons to achieve the purpose of repair, but the effect is limited, the cost is high, and it is easy to cause secondary pollution. its application. 3. Contents of the invention

[0003] In view of this, the present invention provides a compound microbial agent for degra...

Examples

preparation example Construction

[0025] The preparation method of the alkane and polycyclic aromatic hydrocarbon degrading composite bacterial agent of the present invention comprises the following steps:

[0026] Step 1: Single-bacteria fermentation

[0027] Adjust the pH value of the liquid fermentation medium A to 7.4-7.6, carry out aseptic treatment according to the conventional operation, insert the table-top Gordonia NR_61 according to the aseptic operation of 8%-10% of the liquid mass, keep the temperature at 25±1°C, and stir Speed ​​280-320rpm, ventilation 0.3-0.5M 3 / min, tank pressure 0.070-0.085MPa, after continuous fermentation for 75-82h, count live bacteria by hemocytometer, the number of bacteria reached 1.0×10 8 Fermentation was terminated when it was above individual / mL,

[0028] The formula of liquid fermentation medium A is as follows:

[0029] Sucrose 15-20g / L, peptone 1.8-2.0g / L, Na 2 HPO 4 1.5-2.0g / L, KH 2 PO 4 3.5-3.8g / L, (NH 4 ) 2 SO 4 4.0-4.5g / L, MgSO 4 ·7H 2 O 0.7-1.0g...

Embodiment 1

[0040] Step 1: Single-bacteria fermentation

[0041] Adjust the pH value of the liquid fermentation medium A to 7.4, carry out aseptic treatment according to the conventional operation, insert the table-top Gordonia NR_61 according to the aseptic operation of 8% liquid mass, keep the constant temperature at 25°C, the stirring speed at 320rpm, and the ventilation rate at 0.50M 3 / min, tank pressure 0.085MPa, after continuous fermentation for 80h, live bacteria were counted by hemocytometer, and the number of bacteria reached 1.0×10 8 Individual / mL to stop the fermentation;

[0042] Adjust the pH value of the liquid fermentation medium B to 7.2, carry out aseptic treatment according to the conventional operation, insert Pseudomonas stutzeri ZJ-7 according to the aseptic operation of 8% liquid mass, keep the temperature at 36°C, stir at 270rpm, and ventilate Volume 0.35M 3 / min, tank pressure 0.035MPa, after continuous fermentation for 70h, live bacteria were counted by hemocyt...

Embodiment 2

[0053] Step 1: Single-bacteria fermentation

[0054] Adjust the pH value of the liquid fermentation medium A to 7.5, carry out aseptic treatment according to the conventional operation, insert the table-top Gordonia NR_61 according to the aseptic operation of 10% of the liquid mass, keep the temperature at 26°C, stir at 300rpm, and ventilate at 0.40M 3 / min, tank pressure 0.075MPa, after continuous fermentation for 75h, live bacteria were counted by hemocytometer, and the number of bacteria reached 1.0×10 8 Fermentation was terminated when it was more than individual / mL;

[0055] Adjust the pH value of the liquid fermentation medium B to 7.0, carry out aseptic treatment according to the conventional operation, insert Pseudomonas stutzeri ZJ-7 according to the aseptic operation of 6% liquid mass, keep the temperature at 35°C, stir at 250rpm, and ventilate Volume 0.25M 3 / min, tank pressure 0.055MPa, after continuous fermentation for 72h, live bacteria were counted by hemocyto...

1. Technical field:

[0001] The invention relates to a compound bacterial agent for degrading alkanes and polycyclic aromatic hydrocarbons and a preparation method thereof. 2. Background technology:

[0002] Alkanes and polycyclic aromatic hydrocarbons are common toxic, harmful and refractory organic pollutants that seriously endanger the ecological environment and human health. Oil spills, incomplete combustion of fuels such as coal and petroleum, farmland sewage irrigation, and atmospheric deposition are the main sources of them. . Physicochemical methods (chemical oxidation method, thermodynamic repair method, isolation method, photocatalytic method, etc.) can treat alkanes and polycyclic aromatic hydrocarbons to achieve the purpose of repair, but the effect is limited, the cost is high, and it is easy to cause secondary pollution. its application. 3. Contents of the invention

[0003] In view of this, the present invention provides a compound microbial agent for degra...

Examples

preparation example Construction

[0025] The preparation method of the alkane and polycyclic aromatic hydrocarbon degrading composite bacterial agent of the present invention comprises the following steps:

[0026] Step 1: Single-bacteria fermentation

[0027] Adjust the pH value of the liquid fermentation medium A to 7.4-7.6, carry out aseptic treatment according to the conventional operation, insert the table-top Gordonia NR_61 according to the aseptic operation of 8%-10% of the liquid mass, keep the temperature at 25±1°C, and stir Speed ​​280-320rpm, ventilation 0.3-0.5M 3 / min, tank pressure 0.070-0.085MPa, after continuous fermentation for 75-82h, count live bacteria by hemocytometer, the number of bacteria reached 1.0×10 8 Fermentation was terminated when it was above individual / mL,

[0028] The formula of liquid fermentation medium A is as follows:

[0029] Sucrose 15-20g / L, peptone 1.8-2.0g / L, Na 2 HPO 4 1.5-2.0g / L, KH 2 PO 4 3.5-3.8g / L, (NH 4 ) 2 SO 4 4.0-4.5g / L, MgSO 4 ·7H 2 O 0.7-1.0g...

Embodiment 1

[0040] Step 1: Single-bacteria fermentation

[0041] Adjust the pH value of the liquid fermentation medium A to 7.4, carry out aseptic treatment according to the conventional operation, insert the table-top Gordonia NR_61 according to the aseptic operation of 8% liquid mass, keep the constant temperature at 25°C, the stirring speed at 320rpm, and the ventilation rate at 0.50M 3 / min, tank pressure 0.085MPa, after continuous fermentation for 80h, live bacteria were counted by hemocytometer, and the number of bacteria reached 1.0×10 8 Individual / mL to stop the fermentation;

[0042] Adjust the pH value of the liquid fermentation medium B to 7.2, carry out aseptic treatment according to the conventional operation, insert Pseudomonas stutzeri ZJ-7 according to the aseptic operation of 8% liquid mass, keep the temperature at 36°C, stir at 270rpm, and ventilate Volume 0.35M 3 / min, tank pressure 0.035MPa, after continuous fermentation for 70h, live bacteria were counted by hemocyt...

Embodiment 2

[0053] Step 1: Single-bacteria fermentation

[0054] Adjust the pH value of the liquid fermentation medium A to 7.5, carry out aseptic treatment according to the conventional operation, insert the table-top Gordonia NR_61 according to the aseptic operation of 10% of the liquid mass, keep the temperature at 26°C, stir at 300rpm, and ventilate at 0.40M 3 / min, tank pressure 0.075MPa, after continuous fermentation for 75h, live bacteria were counted by hemocytometer, and the number of bacteria reached 1.0×10 8 Fermentation was terminated when it was more than individual / mL;

[0055] Adjust the pH value of the liquid fermentation medium B to 7.0, carry out aseptic treatment according to the conventional operation, insert Pseudomonas stutzeri ZJ-7 according to the aseptic operation of 6% liquid mass, keep the temperature at 35°C, stir at 250rpm, and ventilate Volume 0.25M 3 / min, tank pressure 0.055MPa, after continuous fermentation for 72h, live bacteria were counted by hemocyto...