生态环境学报 ›› 2023, Vol. 32 ›› Issue (3): 619-626.DOI: 10.16258/j.cnki.1674-5906.2023.03.019

• 研究论文 • 上一篇    下一篇

砷氧化菌Pseudomonas sp. AO-1的分离鉴定及其对As(Ⅲ)的氧化性能研究

杨宇(), 邓仁健*(), 隆佩, 黄中杰, 任伯帜, 王政华   

  1. 湖南科技大学土木工程学院，湖南 湘潭 411201
  • 收稿日期:2022-11-21 出版日期:2023-03-18 发布日期:2023-06-02
  • 通讯作者: *邓仁健（1980年生），男，教授，博士，研究方向为重金属废水处理理论与技术。E-mail: deng800912@163.com
  • 作者简介:杨宇（1997年生），男，硕士研究生，研究方向为重金属污染微生物修复治理技术。E-mail: 572965290@qq.com
  • 基金资助:
    湖南省自然科学基金项目(2022JJ30248);湖南省自然科学基金项目(2021JJ30243);湖南省教育厅科学研究项目(20K052);国家自然科学基金项目(41973078);湖南省环保科研课题(HBKT-2022012);湖南省重点研发计划(2022SK2073)

Isolation and Identification of Arsenic-oxidizing Bacterium Pseudomonas sp. AO-1 and Its Oxidation Properties for As(Ⅲ)

YANG Yu(), DENG Renjian*(), LONG Pei, HUANG Zhongjie, Ren Bozhi, WANG Zhenghua   

  1. School of Civil Engineering, Hunan University of Science and Technology, Xiangtan 411201, P. R. China
  • Received:2022-11-21 Online:2023-03-18 Published:2023-06-02

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摘要：

随着环境砷污染日趋严重和不断地传播使得人类健康处于高风险之中，修复治理环境中的砷污染刻不容缓。将As(Ⅲ) 氧化为As(Ⅴ) 是治理砷污染的关键步骤，因此寻找一种经济且绿色的氧化技术成为了研究热点，而微生物氧化As(Ⅲ) 被认为是治理砷污染一种绿色经济可行的方法。从湖南锡矿山矿区含砷的土壤中筛选出一株砷氧化菌，并利用分子生物学技术对其进行了鉴定；研究了其对As(Ⅲ) 的抗性，不同As(Ⅲ) 浓度下的生长特征曲线，不同pH条件下As(Ⅲ) 的氧化性能，生长曲线及其氧化动力学。研究结果表明：该菌属于假单胞菌属（Pseudomonas），将其命名为Pseudomonas sp. AO-1（简称：AO-1），其对As(Ⅲ) 的抗性达2000 mg·L-1，具有较高的抗性；当As(Ⅲ) 质量浓度高于100 mg·L-1时，菌株AO-1的生长受到了明显的抑制，其进入对数增殖期的时间延后，稳定期较短，衰亡期提前；pH对菌株AO-1的生长及As(Ⅲ) 氧化率有显著的影响，其最适生长pH为7；当pH为3-8时，菌株AO-1氧化As(Ⅲ) 的氧化率随pH的升高先提高后减小，pH为7时As(Ⅲ) 氧化率到达最大的72.5%；菌株AO-1好氧化氧化As(Ⅲ) 的米门常数Km和最大氧化速率Vmax值分别为274.70 μmol·L-1和0.31 μmol·(L·min)-1，其Km低于一些已研究的菌株，体现菌株AO-1对As(Ⅲ) 具有良好的亲和力，也表明了其具有较好的As(Ⅲ) 氧化性能。综上，该研究所筛选的菌株AO-1具有良好的耐砷性和As(Ⅲ) 氧化性，在水体及土壤砷污染修复与治理中具有的潜在应用价值。

关键词: 砷污染, 砷氧化菌, Pseudomonas sp. AO-1, 砷抗性, 氧化性能, 氧化动力学

Abstract:

The increasingly serious and continuous spread of arsenic pollution in the environment put human health at high risk, so the remediation and treatment of arsenic pollution are urgent. The oxidation of As(Ⅲ) to As(Ⅴ) is a key step of arsenic pollution control, so the search for economic and green oxidation technology has become a research focus, and microbial oxidation of As(Ⅲ) is considered to be a green and feasible method for arsenic pollution control. In this study, a strain of arsenic oxidizing bacteria was screened from arsenic-containing soil in Xikuangshan mining area of Hunan Province and identified by molecular biological techniques. The resistance of arsenic oxidizing bacteria to As(Ⅲ), the growth characteristic curve at different As(Ⅲ) concentrations, the oxidation performance, growth curve and oxidation kinetics of As(Ⅲ) at different pH conditions were studied. The results showed that the arsenic-oxidizing bacteria belonged to the genus Pseudomonas, which was named as Pseudomonas sp.AO-1 (abbreviated: AO-1), and had high resistance to As(Ⅲ), up to 2000 mg·L-1. When the concentration of As(Ⅲ) was higher than 100 mg·L-1, the growth of strain AO-1 was significantly inhibited, the time to enter the logarithmic growth stage was delayed, the stable stage was shortened, and the decline stage was advanced. The growth of strain AO-1 and the oxidation rate of As(Ⅲ) were significantly affected by pH, and the oxidation rate of strain AO-1 to As(Ⅲ) increased with the increase of pH at 3-7. The optimum growth pH of AO-1 was 7, under which the optimal oxidation of As(Ⅲ) was achieved, and the maximum oxidation rate of As(Ⅲ) reached 72.5%. The Michaelis-Menten constant Km and maximum oxidation rate Vmax of As(Ⅲ) oxidized by strain AO-1 were 274.7008 mol·L-1 and 0.306 mol·(L·min)-1, respectively. And the Km was lower than that of some studied strains, indicating that AO-1 had a good affinity for As(Ⅲ), which also indicated that AO-1 had a good oxidation ability for As(Ⅲ). The strain AO-1 screened in this study showed good arsenic resistance and As(Ⅲ) oxidation, which laid a theoretical foundation for its potential application in the remediation of arsenic polluted water and soil.

Key words: arsenic pollution, arsenic oxidizing bacteria, Pseudomonas sp. AO-1, arsenic resistance, oxidation properties, oxidation kinetics

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