低阶煤的微生物降解在常温、常压下进行，具有能耗低、设备简单和环境友好等优点，是低阶煤绿色转化的有效途径之一。然而，大多数研究发现煤的微生物降解率普遍较低，因此，研究者们尝试利用表面活性剂来优化降解过程，主要研究集中于单一表面活性剂对煤表面性质的改性，而关于组合表面活性剂对微生物细胞的影响及其协同作用促进微生物降解煤的研究较少。

本文选取了2类8种表面活性剂，第一类为化学表面活性剂共6种，包括2种阴离子表面活性剂十二烷基苯磺酸钠（LAS）和十二烷基硫酸钠（SDS），2种阳离子表面活性剂十二烷基三甲基溴化铵（DTAB）和十六烷基氯化吡啶（CPC），2种非离子表面活性剂曲拉通X-100（TR）和吐温80（TW），第二类为生物表面活性剂共2种，分别为鼠李糖脂（RH）和皂素（SA）。研究了单一表面活性剂和组合表面活性剂对茫崖诺卡氏菌降解新疆五彩湾低阶煤降解率和降解时间的影响，再基于组合表面活性剂对细菌生长特性（生物量、酶活和疏水性）、煤样表面性质（接触角、Zeta电位和吸附率）的影响，以及最优组合表面活性剂作用下产物的分布三个方面探讨组合表面活性剂作用下茫崖诺卡氏菌对五彩湾煤降解机理研究，为表面活性剂在煤炭微生物降解中的应用提供了理论依据。

主要工作如下：

（1）茫崖诺卡氏菌降解五彩湾煤最优工艺条件的探索

探索了煤浆浓度、菌液用量、降解时间、煤样粒径和培养方式五个因素对茫崖诺卡氏菌降解五彩湾煤的影响，获得了茫崖诺卡氏菌降解五彩湾煤的最佳降解工艺条件，具体为：煤浆浓度为0.3 g/mL，菌液用量为8 mL/50mL，降解时间为10 d，煤样粒径为0.125-0.250 mm，培养方式为摇床培养，此时A450值最高为0.54，降解率为53.2%。

（2）组合表面活性剂对茫崖诺卡氏菌降解五彩湾煤的影响研究

在最优工艺条件下，探究了单一表面活性剂对降解效果的影响，结果表明表面活性剂DTAB、CPC、SDS和SA均有抑制作用；而表面活性剂TR、TW、LAS和RH有促进作用。当表面活性剂TR、TW、LAS和RH的浓度分别为1200，1200，800和1000 mg/L时，此时降解率最高分别为65.3%，60.6%，56.7%和63.3%。基于表面活性剂对煤炭微生物降解具有促进作用为基本原则，同时兼顾不同类型表面活性剂的协同作用，组合了两类四种组合表面活性剂，一类为非离子-阴离子组合表面活性剂TW-LAS和TR-LAS，另一类为非离子-生物组合表面活性剂TW-RH和TR-RH。

探究了组合表面活性剂对茫崖诺卡氏菌降解五彩湾煤的降解率和降解时间影响，结果表明当非离子-阴离子组合表面活性剂TW-LAS和TR-LAS的配比分别为1200 (6:4) mg/L和1000 (6:4) mg/L时，此时降解率最高分别为69.5%和72.8%，降解时间均为8天。当非离子-生物组合表面活性剂TW-RH和TR-RH的配比分别为1000 (4:6) mg/L和800 (4:6) mg/L时，此时降解率最高，分别为73.6%和78.2%；降解时间均为8天，以上结果表明最优组合表面活性剂为TR-RH。与无表面活性剂相比，在最优组合表面活性剂TR-RH作用下降解率提高了27%，降解时间缩短了2天。

（3）组合表面活性剂对细菌生长特性和五彩湾煤表面性质影响研究

探究了TW-LAS、TR-LAS、TW-RH和TR-RH组合表面活性剂对茫崖诺卡氏菌的生物量、酶活、疏水性、通透性、表面元素和官能团的影响，结果发现四种组合表面活性剂对细菌生长特性影响不同。在最优组合表面活性剂TR-RH作用下，细菌的生物量最大，OD₆₀₀为2.801，碱性蛋白酶的活性最高为102.34 U/mL，细胞的疏水性最强为47.1%，细胞的通透性最大。X射线光电子能谱和红外光谱分析表明在组合表面活性剂TR-RH作用下茫崖诺卡氏菌表面的氮、氧元素含量、-OH和-CH₂官能团增加。这些结果表明在TR-RH作用下，茫崖诺卡氏菌的生长和其分泌碱性蛋白酶活性增加的更为显著，有效地提高了煤炭与茫崖诺卡氏菌或者碱性蛋白酶的接触，进而提高降解率。

同时，探究了TW-LAS、TR-LAS、TW-RH和TR-RH组合表面活性剂对煤样接触角、Zeta电位和吸附率的影响，结果表明四种组合表面活性剂均可以提高煤样的亲水性，增加煤样的Zeta电位和吸附率。与组合表面活性剂TW-LAS、TR-LAS和TW-RH相比，在TR-RH作用下煤样表面的电荷更多，亲水性更强，更有利于煤炭对茫崖诺卡氏菌的吸附。

（4）最优组合表面活性剂作用下降解产物的分布规律及降解机理研究

在最优组合表面活性剂TR-RH作用下降解产物类型与不添加表面活性剂的产物相似，主要含有芳香烃、烷烃、含硫和含氧化合物等，其中烷烃的含量高达90.82%，比不添加表面活性剂提高了16.59%。固相产物结构分析表明在组合表面活性剂TR-RH作用后，煤样中碳含量降低，氧和氮含量升高，芳香度降低，平均缩合环升高。茫崖诺卡氏菌断裂了五彩湾煤中的C-C和C-H键，煤样的热稳定性增加，煤样的孔隙结构被破坏，平均孔径增大。

综上，组合表面活性剂TR-RH不仅可以被茫崖诺卡氏菌作为碳源，进行生理代谢活动，促进自身繁殖和碱性蛋白酶活性，使更多的茫崖诺卡氏菌和碱性蛋白酶与煤炭接触，而且组合表面活性剂可以增加煤样表面的电荷和亲水性，进而增加茫崖诺卡氏菌在煤样表面的吸附，提高降解率。

The biodegradation of low rank coal can be carried out at room temperature and pressure, with the advantages of low energy consumption, simple equipment, and environmental friendliness, making it an effective way for green conversion of low rank coal. However, most studies have found that the biodegradation rate of coal is generally low. Therefore, researchers have attempted to optimize the degradation process by single surfactants. However, there is little research on the effect of combined surfactants on microbial cells and their synergistic effect in promoting biodegradation of coal.

This article selected two categories of eight surfactants. The first category includes six chemical surfactants, including two anionic surfactants sodium dodecyl benzene sulfonate (LAS) and sodium dodecyl sulfate (SDS), two cationic surfactants dodecyl trimethyl ammonium bromide (DTAB) and cetylpyridinium chloride (CPC), and two nonionic surfactants Triton X-100 (TR) and Tween 80 (TW). The second category includes two bio-surfactants, rhamnolipid (RH) and saponin (SA). The study investigated the effects of single surfactants and combined surfactants on the degradation rate and degradation time of the Wucaiwan low rank coal from Xinjiang by the Nocardia mangyaensis (N. mangyanensis). Based on the effects of combined surfactants on bacterial growth characteristics (biomass, enzyme activity, and hydrophobicity), coal surface properties (contact angle, Zeta potential, and adsorption rate), and bioproduct distribution, the mechanism of surfactants in coal biodegradation was explored. This study provides a theoretical basis for the application of surfactants in coal biodegradation.

The main works are as follows:

(1) Study on the optimal process conditions for the degradation of Wucaiwan coal by N. mangyanensis

This study examined the influence of five factors, namely coal slurry concentration, inoculation quantity, degradation time, coal samples particle size and cultivation method on the biodegradation of the Wucaiwan coal by N. mangyanensis. The optimal conditions for the biodegradation of Wucaiwan coal by N. mangyanensis were determined, which were a coal slurry concentration of 0.3 g/mL, bacterial liquid dosage of 8 mL/50 mL, degradation time of 10 days, coal particle size of 0.125-0.250 mm, and cultivation method of shaking cultivation. Under these conditions, the A450 value was the highest at 0.54, and the degradation rate was 53.2%.

(2) Effects of combined surfactants on the degradation of Wucaiwan coal by N. mangyanensis

Under optimal conditions, the effects of single surfactants on degradation were investigated, and the results showed that surfactants DTAB, CPC, SDS, and SA had inhibitory effects, while surfactants TR, TW, LAS, and RH had promoting effects. When the concentrations of surfactants TR, TW, LAS, and RH were 1200, 1200, 800, and 1000 mg/L, the highest degradation rates were 65.3%, 60.6%, 56.7%, and 63.3%, respectively. Based on the principle that surfactants promote biodegradation of coal, while taking into account the synergistic effects of different types of surfactants, nonionic-anionic surfactants TW-LAS and TR-LAS and nonionic-biological surfactants TW-RH and TR-RH were combined.

This study investigated the effects of combined surfactants on the degradation rate and time of Wucailan low-rank coal by N. mangyanensis. The results showed that the highest degradation rates were achieved when the nonionic-anionic combined surfactants TW-LAS and TR-LAS were used at ratios of 1200 (6:4) mg/L and 1000 (6:4) mg/L, respectively, with degradation rates of 69.5% and 72.8%, and a degradation time of 8 days. When the nonionic-biological combined surfactants TW-RH and TR-RH were used at ratios of 1000 (4:6) mg/L and 800 (4:6) mg/L, respectively, the highest degradation rates were 73.6% and 78.2%, and the degradation time was 8 days for both. These results indicate that the optimal combined surfactant was TR-RH. Compared with the control group without surfactants, the degradation rate increased by 27% and the degradation time was reduced by 2 days under the optimal combined surfactants TR-RH.

(3) Effects of combined surfactants on growth characteristics of bacteria and surface properties of Wucaiwan coal

This study investigated the effects of TW-LAS, TR-LAS, TW-RH, and TR-RH combined surfactants on the biomass, enzyme activity, hydrophobicity, permeability, surface elements, and functional groups of N. mangyanensis. Results showed that the four combined surfactants had different effects on bacterial growth characteristics. Under the optimal combined surfactant TR-RH, the bacterial biomass was the highest with an OD₆₀₀ of 2.801, the alkaline protease activity was the highest at 102.34 U/mL, the cell hydrophobicity was the strongest at 47.1%, and the cell permeability was the largest. XPS and FTIR analysis showed that the content of N and O on the surface of N. mangyanensis, as well as the -OH and -CH₂ functional groups, increased under the action of the combined surfactant TR-RH. These results indicated that the growth of N. mangyanensis and its secretion of alkaline protease activity were more significantly increased under the action of TR-RH, effectively improving the contact between coal and N. mangyanensis or alkaline protease, thereby improving the degradation rate.

The effects of combined surfactants TW-LAS, TR-LAS, TW-RH, and TR-RH on coal contact angle were investigated, Zeta potential, and adsorption rate. The results showed that all four combined surfactants can increase the hydrophilicity of the coal sample and increase the Zeta potential and adsorption rate of the coal sample. Compared with the combined surfactants TW-LAS, TR-LAS, and TW-RH, the surface charge of the coal sample was stronger and more hydrophilic under the action of TR-RH, which was more conducive to the adsorption of coal on N. mangyanensis.

(4) Distribution patterns and degradation mechanism of degradation products under the action of the optimal combined surfactant

The types of degradation products under the action of the optimal combined surfactant TR-RH were similar to those the without surfactant, mainly including aromatic hydrocarbons, alkanes, sulfur-containing compounds, and oxygen-containing compounds. The content of alkanes was as high as 90.82%, which was increased by 16.59% compared to that the without surfactant. In addition, after the action of the combined surfactant TR-RH, the C content in the coal sample decreased, while the O and N content increased, and the aromaticity decreased while the average condensed ring increased. The N. mangyanensis broke the C-C and C-H bonds in the Wucaiwan coal, which increased the thermal stability of the coal sample and destroyed the pore structure of the coal sample, resulting in an increase in the average pore size.

In summary, the combined surfactant TR-RH can not only be used as a carbon source by N. mangyanensis, promoting physiological metabolism, self-reproduction, and alkaline protease activity, allowing more N. mangyanensis and alkaline protease to contact with coal. Moreover, the combined surfactant TR-RH can increase the surface charge and hydrophilicity of coal samples, thereby increasing the adsorption of bacteria on the surface of coal samples and improving the degradation rate.

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