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嗜热异养硝化细菌群落GW7的氮转化途径研究

Research on nitrogen transformation pathways of a thermophilic heterotrophic nitrifying bacterial consortium GW7.

作者信息

Ma Yongqi, Wang Jiali, Zhang Yindi, Guan Wenping, Qi Wenrui, Tai Xisheng, Lin Dong, He Rong, Sun Likun, Zhang Aiwen

机构信息

College of Animal Science and Technology, Gansu Agricultural University, Lanzhou, China.

College of Urban Environment, Lanzhou City University, Lanzhou, China.

出版信息

Front Microbiol. 2025 Jun 30;16:1578865. doi: 10.3389/fmicb.2025.1578865. eCollection 2025.

Abstract

INTRODUCTION

High-temperature heterotrophic nitrifying bacteria play a crucial role in the thermophilic phase of aerobic composting by effectively converting reduced ammonia sources. This process reduces ammonia emissions and contributes to nitrogen fixation, thus showcasing a high potential for application. The study aims to isolate and characterize a high-temperature-tolerant heterotrophic nitrifying bacterial consortium to enhance nitrogen transformation during aerobic composting.

METHODS

An excellent high-temperature-tolerant heterotrophic nitrifying bacterial consortium, designated GW7, was enriched from compost samples at elevated temperatures. The bacterial consortium was cultured under varying conditions to determine optimal cultivation parameters. Response surface methodology (RSM) experiments were conducted to find the best conditions for ammonia and nitrate nitrogen utilization. Enzymatic assays were carried out to measure the specific activities of key enzymes, including glutamine synthetase (GS), glutamate dehydrogenase (GDH), glutamate synthetase (GOGAT), ammonia monooxygenase (AMO), and hydroxylamine oxidoreductase (HAO).

RESULTS

At 55°C, the GW7 consortium demonstrated a utilization efficiency of 79.97% for ammonia nitrogen (NH₄+-N) (400 mg/L) and 21.18% for nitrate nitrogen (NO₃-N) (400 mg/L). Response surface methodology identified the optimal cultivation conditions for GW7 as follows: sodium succinate as the carbon source, a C/N ratio of 15:1, a temperature of 53°C, initial pH of 6, and a rotation speed of 200 r/min. Under these conditions, the NH₄-N utilization efficiency increased to 87.80%. Enzymatic assays showed that the specific activities of GS, GDH, and GOGAT were 0.392 U/mg, 0.926 U/mg, and 0.195 U/mg, respectively. Moreover, the specific activities of AMO and HAO were 1.459 U/mg and 0.701 U/mg, respectively.

DISCUSSION

The GW7 consortium demonstrated excellent nitrogen transformation capabilities, effectively utilizing ammonia nitrogen and contributing to the reduction of nitrogen losses in the aerobic composting process. The high enzymatic activities of key nitrogen-metabolizing enzymes, including AMO and HAO, support its role in heterotrophic nitrification. The proposed nitrogen conversion pathways, including ammonia assimilation, heterotrophic nitrification, and assimilatory nitrate reduction, highlight the versatile nitrogen metabolism of this bacterial consortium. The optimized cultivation conditions further enhance its practical application potential in mitigating nitrogen emissions during composting processes.

摘要

引言

高温异养硝化细菌通过有效转化还原态氨源,在好氧堆肥的嗜热阶段发挥着关键作用。这一过程减少了氨排放并有助于固氮,因此展现出很高的应用潜力。本研究旨在分离和表征一种耐高温异养硝化细菌群落,以增强好氧堆肥过程中的氮转化。

方法

从高温堆肥样品中富集得到一个优良的耐高温异养硝化细菌群落,命名为GW7。在不同条件下培养该细菌群落以确定最佳培养参数。进行响应面法(RSM)实验以找出氨和硝酸盐氮利用的最佳条件。进行酶活性测定以测量关键酶的比活性,包括谷氨酰胺合成酶(GS)、谷氨酸脱氢酶(GDH)、谷氨酸合成酶(GOGAT)、氨单加氧酶(AMO)和羟胺氧化还原酶(HAO)。

结果

在55°C时,GW7群落对氨氮(NH₄⁺-N)(400mg/L)的利用效率为79.97%,对硝酸盐氮(NO₃-N)(400mg/L)的利用效率为21.18%。响应面法确定GW7的最佳培养条件如下:以琥珀酸钠作为碳源,碳氮比为15:1,温度为53°C,初始pH为6,转速为200r/min。在这些条件下,NH₄-N的利用效率提高到87.80%。酶活性测定表明,GS、GDH和GOGAT的比活性分别为0.392U/mg、0.926U/mg和0.195U/mg。此外,AMO和HAO的比活性分别为1.459U/mg和0.701U/mg。

讨论

GW7群落展现出优异的氮转化能力,能有效利用氨氮并有助于减少好氧堆肥过程中的氮损失。包括AMO和HAO在内的关键氮代谢酶的高酶活性支持了其在异养硝化中的作用。所提出的氮转化途径,包括氨同化、异养硝化和同化性硝酸盐还原,突出了该细菌群落多样的氮代谢。优化后的培养条件进一步增强了其在减轻堆肥过程中氮排放方面的实际应用潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a641/12257491/60a0010d24fd/fmicb-16-1578865-g001.jpg

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