煤炭的微生物转化在常温、常压下进行，具有耗能低、设备要求简单的优点，是一种煤炭绿色利用的新途径。目前，人们对煤炭生物降解的研究已取得一定成果，但研究主要集中于褐煤的微生物降解，对于富油煤的微生物降解及其降解产生的生物腐殖酸的研究尚处于初始阶段。

本文选择荧光假单胞菌和铜绿假单胞菌两种细菌降解陕北郭家沟、锦界、上湾三种富油煤，探究了两种细菌降解三种富油煤的最优工艺条件，在最优工艺条件下从降解产物中分离得到六种生物腐殖酸，通过碱抽提法从三种富油煤中制取得到三种化学腐殖酸作对照组，研究这九种腐殖酸在结构和性能方面的差异，并探究其对蔬菜上海青生长的影响。

主要工作如下：

(1) 荧光假单胞菌降解三种陕北富油煤制取生物腐殖酸最优工艺条件探索

荧光假单胞菌降解郭家沟富油煤、锦界富油煤和上湾富油煤制取腐殖酸的最佳工艺条件分别为煤浆浓度0.5 g/50 mL，降解时间14 d，菌液用量12 mL/50 mL、煤浆浓度0.3 g/50 mL，降解时间14 d，菌液用量16 mL/50 mL、煤浆浓度0.3 g/50 mL，降解时间12 d，菌液用量6 mL/50 mL，对应腐殖酸的最高制取率为32.87%、21.31%、66.96%。

(2) 铜绿假单胞菌降解三种陕北富油煤制取生物腐殖酸最优工艺条件探索

铜绿假单胞菌降解郭家沟富油煤、锦界富油煤和上湾富油煤制取腐殖酸的最佳工艺条件分别为煤浆浓度0.3 g/50 mL，降解时间8 d，菌液用量6 mL/50 mL、煤浆浓度0.3 g/50 mL，降解时间10 d，菌液用量12 mL/50 mL、煤浆浓度0.3 g/50 mL，降解时间8 d，菌液用量8 mL/50 mL，对应腐殖酸的最高制取率为39.33%、30.85%、75.88%。

(3) 生物腐殖酸与化学腐殖酸在结构和性能上的对比分析

通过碱抽提法对郭家沟富油煤、锦界富油煤和上湾富油煤进行提取得到三种化学腐殖酸，分析六种生物腐殖酸和三种化学腐殖酸在结构方面的差异发现：生物腐殖酸含有脂肪族链较多，低分子化合物较多，含氧官能团少，晶面间距大，微晶高度小，碳层数少，整体微晶结构尺寸较小，经二氯甲烷萃取后九种腐殖酸中所含物质种类相似度较高，都是以烷烃类为主，还含有一些芳香类、醇类、酯类、羧酸类、醚类及胺类物质，但其含量有所差别。有机质元素分析发现生物腐殖酸含有远高于化学腐殖酸的有机质元素，使得生物腐殖酸比化学腐殖酸作为农作物营养源更具优势。

(4) 生物腐殖酸与化学腐殖酸对蔬菜上海青生长的影响

两类腐殖酸与不加腐殖酸的空白组相比，加入两类腐殖酸后对蔬菜上海青发芽率、茎粗、高度、重量均有较大促进作用，生物腐殖酸和化学腐殖酸在对蔬菜上海青茎粗、高度、重量的长势方面促进作用相当，但在促进蔬菜上海青的发芽率方面生物腐殖酸更具优势，尤其是荧光假单胞菌降解煤样后产生的腐殖酸在上海青种植后的前4天促进作用更强，这可能与荧光假单胞菌降解后制取的腐殖酸分子量小有关系。

The microbial conversion of coal is carried out at normal temperature and pressure, which has the advantages of low energy consumption and simple equipment requirements, and is a new way of green utilization of coal. At present, people have made certain achievements in the study of biological degradation of coal, but the research mainly focuses on the microbial degradation of lignite. The study on the microbial degradation of rich oil coal and the production of humic acid by its degradation is still in the initial stage.

The study selected two types of bacteria, Pseudomonas fluorescens and Pseudomonas aeruginosa, for the degradation of three types of high-oil coal from Guojiagou, Jinjie, and Shangwan in Shaanbei. The research aimed to explore the optimal conditions for the bacterial degradation of the three types of high-oil coal. Under these optimal conditions, six types of bio-humic acids were isolated from the degradation products. Additionally, three types of chemical humic acids were extracted from the three types of high-oil coal using an alkaline extraction method as a control group. The study investigated the differences in structure and properties among these nine types of humic acids and explored their impact on the growth of Shanghaiqing vegetables.

The main work is as follows:

(1) Exploration of optimal conditions for the production of biologically-derived humic acids by Pseudomonas fluorescens during the degradation of three types of rich oil coal from northern Shaanxi.

The optimal conditions for the production of humic acids by the degradation of Guojiagou, Jinjie, and Shangwan rich oil coal using Pseudomonas fluorescens are as follows: coal slurry concentration of 0.5 g/50 mL, degradation time of 14 days, and a bacterial liquid dosage of 12 mL/50 mL for Guojiagou rich oil coal; coal slurry concentration of 0.3 g/50 mL, degradation time of 14 days, and a bacterial liquid dosage of 16 mL/50 mL for Jinjie rich oil coal; coal slurry concentration of 0.3 g/50 mL, degradation time of 12 days, and a bacterial liquid dosage of 6 mL/50 mL for Shangwan rich oil coal. The corresponding maximum yields of humic acid are 32.87%, 21.31%, and 66.96%, respectively.

(2) Exploration of optimal conditions for the productions of biologically-derived humic acids by Pseudomonas aeruginosa during the degradations of three types of rich oil coal from northern Shaanxi.

The optimal conditions for the production of humic acids derived from the degradation of Guojiagou, Jinjie, and Shanwan rich oil coal by Pseudomonas aeruginosa were found to be a coal slurry concentration of 0.3 g/50 mL, a degradation time of 8 days, and a bacterial liquid volume of 6 mL/50 mL for Guojiagou coal; a coal slurry concentration of 0.3 g/50 mL, a degradation time of 10 days, and a bacterial liquid volume of 12 mL/50 mL for Jinjie coal; and a coal slurry concentration of 0.3 g/50 mL, a degradation time of 8 days, and a bacterial liquid volume of 8 mL/50 mL for Shanwan coal, with the corresponding highest humic acid yield of 39.33%, 30.85%, and 75.88%, respectively.

(3) The comparative analysis between the structures and properties of biological humic acid and chemical humic acid.

Three types of humic acids were extracted from Guojia Gou rich oil coal, Jinjie rich oil coal, and Shangwan rich oil coal through alkali extraction. Structural differences between six types of biological humic acids and three types of chemical humic acids were analyzed. It was found that biological humic acids contained more aliphatic chains, more low-molecular-weight compounds, fewer oxygen functional groups, larger interplanar spacing, lower microcrystal height, fewer carbon layers, and smaller overall microcrystalline structure size. After extraction with dichloromethane, the similarity in substance types among the nine humic acids was high, with alkanes being the main type, and also containing some aromatic, alcohol, ester, carboxylic acid, ether, and amine substances, but their content varied. Organic elemental analysis revealed that biological humic acids contained significantly higher organic matter elements than chemical humic acids, making biological humic acids more advantageous than chemical humic acids as a source of nutrients for crops.

(4) The impacts of bio-humic acid and chemical humic acid on the growth of Shanghai green vegetables.

Compared to the blank control group without humic acid, the addition of the two types of humic acid showed significant promotion in terms of germination rate, stem thickness, height, and weight of Shanghaiqing vegetables. Both bio-humic acid and chemical humic acid demonstrated similar promotion effects on the growth of Shanghaiqing vegetables in terms of stem thickness, height, and weight. However, bio-humic acid had an advantage in promoting the germination rate of Shanghaiqing vegetables, especially the humic acid produced after the degradation of coal by Pseudomonas fluorescens, which showed stronger promotion effects within the first four days. This could be attributed to the smaller molecular weight of humic acid derived from the degradation process of Pseudomonas fluorescens.

Compared to the blank control group without the addition of humic acid, the inclusion of two types of humic acid resulted in significant promotion of germination rate, stem thickness, height, and weight of Shanghaiqing vegetables. Both bio-humic acid and chemical humic acid showed similar promotion effects on the growth of Shanghaiqing vegetables in terms of stem thickness, height, and weight. However, bio-humic acid had an advantage in promoting the germination rate of Shanghaiqing vegetables, especially the humic acid produced after the degradation of coal by Pseudomonas fluorescens, which had a stronger promotion effect within the first four days after Shanghaiqing vegetable planting. This could be attributed to the smaller molecular weight of humic acid derived from the degradation process of Pseudomonas fluorescens.