铁改性生物炭-凹凸棒石载体固定化菌剂制备及其对氯苯污染土壤修复作用

doi:10.16258/j.cnki.1674-5906.2024.11.012

生态环境学报 ›› 2024, Vol. 33 ›› Issue (11): 1782-1791.DOI: 10.16258/j.cnki.1674-5906.2024.11.012

• 研究论文【环境科学】 • 上一篇下一篇

铁改性生物炭-凹凸棒石载体固定化菌剂制备及其对氯苯污染土壤修复作用

李文章¹^,²^,⁵(), 胡亚茹¹^,³^,^*(), 李法云¹^,²^,^*(), 王玮¹^,³, 张继宁⁴, 郭琴¹^,²

1.上海应用技术大学生态技术与工程学院，上海 201418
2.美丽中国与生态文明研究院（上海高校智库），上海 201418
3.上海城市路域生态工程技术研究中心，上海 201418
4.上海市农业科学院生态环境保护研究所，上海 201403
5.上海城建水务工程有限公司，上海 200082

收稿日期:2024-06-21 出版日期:2024-11-18 发布日期:2024-12-06
通讯作者: 李法云。E-mail: lnecology@163.com
*胡亚茹。E-mail: hu746459@163.com;
作者简介:李文章（1997年生），男，硕士研究生，研究方向为有机污染场地修复。E-mail: lwz18333286318@163.com
基金资助:
国家重点研发计划项目(2020YFC1808802);上海市青年科技英才扬帆计划项目(21YF1446500);上海市地方能力建设计划项目(20090503200);上海应用技术大学引进人才科学研究项目(YJ2023-26)

Preparation of Iron Modified Biochar-attapulgite Carrier Immobilized Bacterial Agent and Its Remediation for Soil Contaminated by Chlorobenzene

LI Wenzhang¹^,²^,⁵(), HU Yaru¹^,³^,^*(), LI Fayun¹^,²^,^*(), WANG Wei¹^,³, ZHANG Jining⁴, GUO Qin¹^,²

1. School of Ecological Technology and Engineering, Shanghai Institute of Technology, Shanghai 201418, P. R. China
2. Institute of Beautiful China and Ecological Civilization, University Think Tank of Shanghai Municipality, Shanghai 201418, P. R. China
3. Center for Urban Road Ecological Engineering and Technology of Shanghai Municipality, Shanghai 201418, P. R. China
4. Eco-Environmental Protection Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai, 201403, P. R. China
5. Shanghai Urban Construction Water Engineering Co., LTD., Shanghai 200082, P R. China

Received:2024-06-21 Online:2024-11-18 Published:2024-12-06

摘要/Abstract

摘要：

氯苯是有机化工行业重要的中间体和有机溶剂，经生产、运输过程泄漏以及末端违规排放而迁移至土壤，严重威胁生态环境和人体健康。氯苯对土壤的污染状况及绿色高效修复一直是环境领域的热点问题。微生物修复技术具有能耗低、绿色无二次污染等优点，但其在污染土壤中的实践往往面临菌株存活能力差、降解效率低等关键技术难题。生物炭可作为载体固定化高效降解微生物，但其性能常受到比表面积、官能团等因素制约。将硫酸铁、凹凸棒石和芦苇秸秆以1꞉2꞉10的比例均匀混合后在500 ℃条件下无氧热解2 h，对芦苇生物炭进行改性以提高其对降解微生物的固定能力。进一步优化了载体固定化铜绿假单胞菌Pseudomonas aeruginosa W3的条件，制备铁改性生物炭-凹凸棒石载体固定化菌剂（BFAT@W3），并将其应用于污染土壤修复。结果表明，经硫酸铁和凹凸棒石改性后的生物炭比表面积（210.7 m²·g^-1）和孔体积（0.13 cm³·g^-1）较高，官能团种类丰富。固定化降解微生物的最佳条件为吸附温度37 ℃、吸附时间16 h、载体投加量15 mg·mL^-1，此条件下固定的有效活细胞浓度为2.10×10⁹ cell·g^-1。在实际氯苯污染土壤中，投加比例为3꞉100的BFAT@W3材料对氯苯去除率较高（83.4%），较相同比例的W3提高了18.6%。16S rRNA高通量测序结果显示，3꞉100的BFAT@W3能有效增加土壤中变形菌门（Proteobacteria）和假单胞菌属（Pseudomonas）的相对丰度，相较于同比例的W3处理组分别提高了41.3%和36.7%，增强了降解微生物在土壤群落中的竞争优势。该研究通过制备性能良好的铁改性生物炭-凹凸棒石载体固定化高效降解菌剂，可为氯苯污染土壤的生物修复提供技术支撑。

关键词: 生物炭, 微生物, 固定化菌剂, 氯苯, 生物修复

Abstract:

Chlorobenzene is an important intermediate and an organic solvent used in the organic chemical industry. In the process of production, transportation, and illegal emissions, chlorobenzene migrates into the soil, posing a serious threat to the ecological environment, even directly entering and endangering human health through the food chain. As a significant receiving environment for chlorobenzene, the pollution status and green efficient remediation of soil have always been a hot issue in the environmental field. Microbial remediation has attracted much attention in the field of environmental remediation owing to its advantages of low energy consumption, green production, and lack of secondary pollution. However, microbial remediation is often confronted with key technical problems in the practice of contaminated soil remediation, such as poor survival ability and low degradation efficiency of bacterial strains. Biochar can be used to immobilize microorganisms for efficient degradation; however, its performance is often limited by its specific surface area, functional groups, and other factors. In this study, iron sulfate, attapulgite, and reed straw were mixed at a ratio of 1꞉2꞉10 and then pyrolyzed at a high temperature of 500 ℃ for 2 h without oxygen. Reed biochar was modified to improve its immobilization capacity for degrading microorganisms. Furthermore, the immobilization conditions of Pseudomonas aeruginosa W3 were optimized to prepare an iron-modified biochar-attapulgite carrier-immobilized bacterial agent (BFAT@W3) for the application in the remediation of contaminated soil. The results showed that, after the combined modification with the Fe₂(SO₄)₃ and attapulgite, the specific surface area (210.7 m²·g^-1) and pore volume (0.13 cm³·g^-1) of BC-Fe-ATP were higher and the functional groups were more abundant. The optimal conditions for the immobilization of microorganisms were as follows: adsorption temperature of 37 ℃, the adsorption time of 16 h, and carrier dosage of 15 mg·mL^-1. Under these conditions, the concentration of live cells immobilized on the carrier was 2.10×10⁹·g^-1. In practical soil contaminated by chlorobenzene, the removal rate of chlorobenzene was higher (83.4%), while the proportion of BFAT@W3 was 3꞉100, which was 18.6% higher than that of W3. High-throughput sequencing of 16S rRNA showed that the application of 3꞉100 BFAT@W3 effectively increased the relative abundance of Proteobacteria and Pseudomonas in the soil. Compared with the same proportion of W3, the application of 3꞉100 BFAT@W3 effectively increased the relative abundance of Proteobacteria and Pseudomonas in the soil by 41.3% and 36.7%, respectively, enhancing the competitive advantage in soil microbial communities. In this study, an iron-modified biochar-attapulgite carrier with good performance was used to immobilize efficient degrading bacterial agents, which provided technical support for the bioremediation of chlorobenzene-contaminated soil.

Key words: biochar, microorganisms, immobilized bacterial agent, chlorobenzene, bioremediation

中图分类号:

X53
TQ413

李文章, 胡亚茹, 李法云, 王玮, 张继宁, 郭琴. 铁改性生物炭-凹凸棒石载体固定化菌剂制备及其对氯苯污染土壤修复作用[J]. 生态环境学报, 2024, 33(11): 1782-1791.

LI Wenzhang, HU Yaru, LI Fayun, WANG Wei, ZHANG Jining, GUO Qin. Preparation of Iron Modified Biochar-attapulgite Carrier Immobilized Bacterial Agent and Its Remediation for Soil Contaminated by Chlorobenzene[J]. Ecology and Environment, 2024, 33(11): 1782-1791.

图/表 16

表1 氯苯的理化性质

Table 1 Physicochemical properties of chlorobenzene

名称	分子式	分子量	辛醇-水分配系数的对数值	相对密度 (水꞉1)	熔点/ ℃	沸点/ ℃
1-氯苯	C₆H₅Cl	112.56	2.84	1.10	-45.2	132.2

表2 BFAT@W3染色处理

Table 2 Dyeing treatment of BFAT@W3

组别	SYTO 9	PI
Blank	‒	‒
SYTO 9	2 μL、37 ℃避光培养30 min	‒
SYTO 9+PI	2 μL、37 ℃避光培养30 min	4 μL、37 ℃避光培养30 min
BFAT@W3	2 μL、37 ℃避光培养30 min	4 μL、37 ℃避光培养30 min

表3 MS 培养基主要成分

Table 3 The main components of MS medium

元素成分	相对分子质量	质量浓度/(mg·L^-1)
KNO₃	101.21	1900
NH₄NO₃	80.04	1650
KH₂PO₄	136.09	170
MgSO₄·7H₂O	246.47	370
CaCl₂·2H₂O	147.02	440

表4 BFAT@W3去除氯苯实验设计

Table 4 Experimental design for the removal of chlorobenzene by BFAT@W3

组别	BFAT@W3 投加比例	加入BFAT@W3 质量/g	氯苯溶液体积/mL
CK	0	0	50
T1	1꞉100	0.5	50
T2	2꞉100	1.0	50
T3	3꞉100	1.5	50

表5 BFAT@W3修复污染土壤实验设计

Table 5 Experimental design of BFAT@W3 for remediation of contaminated soil

组别	处理方式
CK	无
BC-Fe-ATP	铁改性生物炭-凹凸棒石复合载体
W3	游离菌
BFAT@W3	复合载体固定化菌剂

表6 3种载体材料的平均孔径、比表面积和总孔体积

Table 6 Average pore size, specific surface area, and total pore volume for the three carrier materials

材料类型	平均孔径/nm	比表面积/(m²·g^-1)	总孔体积/(m³·g^-1)
BC	5.07	23.4	0.03
BC-ATP	8.43	26.5	0.04
BC-Fe-ATP	2.30	210.7	0.13

图1 BC、BC-ATP与BC-Fe-ATP的FTIR图

Figure 1 FTIR image of BC, BC-ATP and BC-Fe-ATP

图2 不同条件下BC-Fe-ATP对W3的固定率

Figure 2 Immobilization rates of W3 by BC-Fe-ATP under different conditions

图3 游离菌、载体及载体固定化菌剂SEM图

Figure 3 SEM images of free bacteria, carriers, and carriers immobilized bacterial agent

图4 不同投加比例的BFAT@W3对氯苯的去除

Figure 4 Removal of chlorobenzene by BFAT@W3 with different proportion of dosag

图5 不同处理组对土壤中氯苯的去除

Figure 5 Removal of chlorobenzene from soil by different treatment groups

表7 修复前后土壤pH、电导率和有机碳含量变化

Table 7 Changes of soil pH, electrical conductivity and organic carbon content before and after restoration

处理组	pH		电导率/(dS·m^-1)		有机碳质量分数/(g·kg^-1)
处理组	修复前	修复后	修复前	修复后	修复前	修复后
CK	7.76±0.05	7.65±0.02	2.33±0.02	2.396±0.01	14.54±0.34	15.14±0.49
W3		7.46±0.04		2.10±0.06		13.63±0.38
BC-Fe-ATP		8.4±0.05		2.914±0.01		18.26±0.39
BFAT@W3		7.74±0.04		2.213±0.04		17.90±0.46

表8 土壤Alpha多样性指数

Table 8 The Alpha diversity indexes of soils

样品	Alpha多样性指数
样品	Shannon	Chao	Ace	Simpson	Coverage	Shannoneven
CK	6.00	2631.02	2723.79	0.03	0.99	0.76
W3	4.83	2314.85	2426.33	0.08	0.99	0.63
BC-Fe-ATP	6.09	2565.87	2656.27	0.03	0.99	0.78
BFAT@W3	2.07	953.49	1017.84	0.4	1.00	0.31

图6 不同处理组土壤微生物群落主坐标分析

Figure 6 Principal co-ordinates Analysis of soil microbial communities in different treatment groups

图7 各处理组群落细菌门水平Heatmap图

Figure 7 Heatmap of bacterial at phylum level in each treatment group in community

图8 各处理组群落细菌属水平Heatmap图

Figure 8 Heatmap of bacterial at genus level in each treatment group in community

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铁改性生物炭-凹凸棒石载体固定化菌剂制备及其对氯苯污染土壤修复作用

Preparation of Iron Modified Biochar-attapulgite Carrier Immobilized Bacterial Agent and Its Remediation for Soil Contaminated by Chlorobenzene

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