邻苯二甲酸二乙基己基酯降解菌株的筛选及其降解特性

郭赫淇; 张文丽; 徐伟慧; 王志刚; 陈文晶; 胡云龙

文章摘要

邻苯二甲酸二乙基己基酯降解菌株的筛选及其降解特性

Isolation of di （2-ethylhexyl）phthalate-degrading strains and their degradation characteristics

投稿时间：2024-08-25

DOI：10.13254/j.jare.2024.0664

中文关键词: DEHP，菌株筛选，微生物降解，共培养，降解途径

英文关键词: DEHP, strain screening, microbial degradation, co-cultivation, degradation pathway

基金项目:黑龙江省自然科学基金杰出青年项目（JQ2023D001）；黑龙江省省属高等学校基本科研业务费（145309401）

作者	单位	E-mail
郭赫淇	齐齐哈尔大学生命科学与农林学院, 黑龙江齐齐哈尔 161006 黑龙江省农业微生物制剂产业化技术创新中心, 黑龙江齐齐哈尔 161006 黑龙江省农用生物制剂产业化协同创新中心, 黑龙江齐齐哈尔 161006
张文丽	齐齐哈尔大学生命科学与农林学院, 黑龙江齐齐哈尔 161006 黑龙江省农业微生物制剂产业化技术创新中心, 黑龙江齐齐哈尔 161006 黑龙江省农用生物制剂产业化协同创新中心, 黑龙江齐齐哈尔 161006
徐伟慧	齐齐哈尔大学生命科学与农林学院, 黑龙江齐齐哈尔 161006 黑龙江省农业微生物制剂产业化技术创新中心, 黑龙江齐齐哈尔 161006 黑龙江省农用生物制剂产业化协同创新中心, 黑龙江齐齐哈尔 161006
王志刚	齐齐哈尔大学生命科学与农林学院, 黑龙江齐齐哈尔 161006 黑龙江省农业微生物制剂产业化技术创新中心, 黑龙江齐齐哈尔 161006 黑龙江省农用生物制剂产业化协同创新中心, 黑龙江齐齐哈尔 161006	wangzhigang@qqhru.edu.cn
陈文晶	齐齐哈尔大学生命科学与农林学院, 黑龙江齐齐哈尔 161006 黑龙江省农业微生物制剂产业化技术创新中心, 黑龙江齐齐哈尔 161006 黑龙江省农用生物制剂产业化协同创新中心, 黑龙江齐齐哈尔 161006
胡云龙	齐齐哈尔大学生命科学与农林学院, 黑龙江齐齐哈尔 161006 黑龙江省农业微生物制剂产业化技术创新中心, 黑龙江齐齐哈尔 161006 黑龙江省农用生物制剂产业化协同创新中心, 黑龙江齐齐哈尔 161006

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中文摘要:

为筛选邻苯二甲酸二乙基己基酯（DEHP）降解菌株，并探究其共培养条件下的相互作用及协同降解特性，本研究取黑龙江省克山县未受DEHP污染的黑土添加40 mg·kg^-1 DEHP设置微宇宙实验，30 d后取处理后黑土以稀释涂布法筛选DEHP降解菌，分离纯化菌株并鉴定；通过扫描电镜观察菌株微观形态对DEHP胁迫的响应；依据菌株对DEHP及下游代谢产物的趋化性、生长适应性的互补特征进行菌株的复配共培养；通过菌株代谢物交叉喂养确定其相互作用，研究菌株共培养降解DEHP的特性。结果表明，筛选得到的两株DEHP降解菌株鉴定为Klebsiella quasivariicola GHQ2和Comamonas testosterone GHQ27，两菌株在400 mg·L^-1 DEHP胁迫下细胞表面出现褶皱，对DEHP及其下游代谢产物均具有趋化性和生长适应性。GHQ27的代谢物可以促进GHQ2的生长，GHQ2-GHQ27共培养较菌株单培养显著提高了生物量。GHQ2-GHQ27共培养72 h内对初始浓度400 mg·L^-1 DEHP的最高降解率可达98.89%，而菌株GHQ2、GHQ27单培养对400 mg·L^-1 DEHP的降解率分别为90.72%、93.39%。根据DEHP降解过程中检测的代谢产物推测，GHQ2-GHQ27共培养通过β氧化和脱酯化双途径共同作用，实现DEHP完全降解。GHQ2-GHQ27共培养具备良好的DEHP降解能力，具有进一步开发为菌剂的潜力。本研究结果为后续DEHP污染土壤修复菌剂开发提供理论依据和技术支持。

英文摘要:

To screen for bacterial strains that degrade di（2-ethylhexyl） phthalate（DEHP）, and to investigate the interactions and synergistic degradation characteristics of the two strains under co-culture conditions. A microcosm experiment was established by adding 40 mg·kg^-1 DEHP to black soil not contaminated by DEHP from Keshan County, Heilongjiang Province. After 30 days, the treated soil was taken and dilution-plating method was used to isolate and identify DEHP-degrading bacteria, and the strains were isolated, purified and identified; the microscopic morphology of the strains in response to DEHP stress was examined using scanning electron microscopy; two strains were co-cultivated based on their complementary characteristics in chemotaxis towards DEHP and its downstream metabolites, as well as their growth adaptability; and the microbial interactions were determined through cross-feeding of metabolites to facilitate the study of DEHP degradation characteristics in co-cultures. Two DEHP-degrading bacterial strains were screened out and identified as Klebsiella quasivariicola GHQ2 and Comamonas testosterone GHQ27. Under stress of 400 mg·kg^-1 DEHP, both strains had surface wrinkling and demonstrated chemotactic responses as well as growth adaptability to DEHP and its downstream metabolites. The study demonstrated that the metabolites of GHQ27 could promote the growth of GHQ2. Co-cultivation of GHQ2 and GHQ27 significantly increased biomass compared with individual culture of the strains. We observed that the co-cultivation of GHQ2 and GHQ27 achieved a maximum degradation rate of 98.89% for DEHP with an initial concentration of 400 mg·L^-1 within 72 h. In contrast, the individual strains GHQ2 and GHQ27 had degradation rates of 90.72% and 93.39%, respectively. Based on the metabolites detected during the degradation of DEHP, we hypothesize that the co-cultivation of GHQ2 and GHQ27 achieves complete degradation of DEHP through the combined pathways of β-oxidation and de-esterification. Through the co-cultivation of GHQ2 and GHQ27, DEHP can be degraded and further developed into bacteria agent, providing a theoretical basis and technical support for the subsequent development of DEHP-contaminated soil remediation bacteria agent.

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