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基于 Taguchi 实验设计优化新分离的枯草芽孢杆菌 AGM5 对马拉硫磷的降解潜力并阐明其降解途径。

Optimizing the malathion degrading potential of a newly isolated Bacillus sp. AGM5 based on Taguchi design of experiment and elucidation of degradation pathway.

机构信息

Department of Environmental Science, School of Earth Sciences, Central University of Rajasthan, BandarSindri, Ajmer, Rajasthan, 305817, India.

出版信息

Biodegradation. 2022 Oct;33(5):419-439. doi: 10.1007/s10532-022-09986-x. Epub 2022 May 16.

DOI:10.1007/s10532-022-09986-x
PMID:35575957
Abstract

Malathion, a pesticide used to control pests in crops, vegetables, fruits, and livestock. Its widespread and indiscriminate usage has ensued in different ecological issues, thus, it's vital to remediate this insecticide. Malathion degrading bacterium Bacillus sp. AGM5, isolated from pesticide contaminated agricultural field was cultured in presence of different malathion concentrations under aerobic and energy restrictive conditions and was found effective at malathion degradation. Recovered malathion was extracted based on QuEChERS approach and then analyzed by UHPLC. About 39.5% of malathion biodegradation was observed at 300 µlL after 96 h of incubation with the tested bacteria which increased to 58.5% and 72.5% after 240, and 360 h of incubation, respectively. To further enhance malathion biodegradation, the effects of co-substrates, pH, temperature, initial malathion concentration, agitation (rpm), and inoculum size were evaluated using Taguchi methodology. Taguchi DOE's ability to predict the optimal response was established experimentally via optimised levels of these factors (glucose-0.1%, yeast extract-0.1%, inoculum size-2% wv, malathion concentration 300 µlL, rpm-150, pH-7, temperature 40 °C), whereby biodegradation rate was enhanced to 95.49% after 38 h. Confirmation of malathion biodegradation was performed by UHPLC, Q-TOF-MS, GC-MS analysis and a possible degradation pathway was proposed for malathion biodegradation. First order kinetic model was appropriate to describe malathion biodegradation. The Taguchi DOE proved to be viable tool for optimizing culture conditions and analysing the interactions between process parameters in order to attain the best feasible combination for maximum malathion degradation. These results could influence the development of a bioremediation strategy.

摘要

马拉硫磷是一种用于控制作物、蔬菜、水果和牲畜中害虫的农药。它的广泛和无差别使用导致了不同的生态问题,因此,修复这种杀虫剂至关重要。从受农药污染的农田中分离出的降解菌芽孢杆菌 sp. AGM5,在有氧和能量限制条件下,在不同马拉硫磷浓度下培养,被发现对马拉硫磷降解有效。回收的马拉硫磷基于 QuEChERS 方法提取,然后通过 UHPLC 分析。在 96 小时的孵育后,用测试细菌观察到 39.5%的马拉硫磷生物降解,分别在 240、360 小时的孵育后,增至 58.5%和 72.5%。为了进一步提高马拉硫磷的生物降解能力,采用 Taguchi 法评估了共底物、pH 值、温度、初始马拉硫磷浓度、搅拌(rpm)和接种量的影响。通过这些因素的最佳水平(葡萄糖-0.1%、酵母提取物-0.1%、接种量-2%wv、马拉硫磷浓度 300 μlL、rpm-150、pH-7、温度 40°C)的实验优化,证明了 Taguchi DOE 预测最佳响应的能力,生物降解率在 38 小时后提高到 95.49%。通过 UHPLC、Q-TOF-MS、GC-MS 分析确认马拉硫磷生物降解,并提出马拉硫磷生物降解的可能途径。一级动力学模型适合描述马拉硫磷的生物降解。Taguchi DOE 被证明是一种可行的工具,可用于优化培养条件,并分析工艺参数之间的相互作用,以获得最大马拉硫磷降解的最佳可行组合。这些结果可能会影响生物修复策略的发展。

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