克雷伯氏菌(Klebsiella)Z3的分离鉴定和对铬(Ⅵ)还原特性研究
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摘要
从河南某矿渣堆放区土壤中筛选到1株高效还原Cr(Ⅵ)的菌株,经形态观察、生理生化特性鉴定以及16S rRNA核酸序列系统发育分析,该菌株为克雷伯氏菌(Klebsiella),命名为Z3。抗逆分析表明,Z3是1株嗜碱耐盐的好氧菌。通过正交优化试验确定Z3还原Cr(Ⅵ)的最适温度为36℃,最适p H值为9. 0,对70 mg·L-1Cr(Ⅵ)在24 h的还原率高达92%。Cr(Ⅵ)还原动力学分析表明,在静息条件下,Z3对Cr(Ⅵ)的还原呈现零级反应动力学特征;在生长状态下,Z3对Cr(Ⅵ)的还原呈现一级反应动力学特征。
A highly efficient Cr( Ⅵ) reducing strain was screened from the soil of a slag stacking area in Henan province. Through morphological observation,identification of physiological and biochemical characteristics,16 S rRNA nucleic acid sequence and phylogenetic ansysis,the strain was determined as Gram-negative Bacillus and identified as Klebsiella sp. Stress resistance analysis showed that,Z3 strains was on obligate alkaliphilic and halotolerant and aerobic bacterium. Orthogonal experiment was further designed to get the Cr( Ⅵ) reduction conditions by strain Z3. The optimal temperature and the initial pH value of medium were 30 ℃ and 9. 0,respectively. The reduction rate of Cr( Ⅵ) at the concentration of 70 mg· L-1 was 92% within 24 hours. The kinetic analysis of Cr( Ⅵ) reduction showed that the reduction of Cr( Ⅵ) by Z3 was a zero-order reaction under the resting condition and was a first-order reaction in the growth state.
A highly efficient Cr( Ⅵ) reducing strain was screened from the soil of a slag stacking area in Henan province. Through morphological observation,identification of physiological and biochemical characteristics,16 S rRNA nucleic acid sequence and phylogenetic ansysis,the strain was determined as Gram-negative Bacillus and identified as Klebsiella sp. Stress resistance analysis showed that,Z3 strains was on obligate alkaliphilic and halotolerant and aerobic bacterium. Orthogonal experiment was further designed to get the Cr( Ⅵ) reduction conditions by strain Z3. The optimal temperature and the initial pH value of medium were 30 ℃ and 9. 0,respectively. The reduction rate of Cr( Ⅵ) at the concentration of 70 mg· L-1 was 92% within 24 hours. The kinetic analysis of Cr( Ⅵ) reduction showed that the reduction of Cr( Ⅵ) by Z3 was a zero-order reaction under the resting condition and was a first-order reaction in the growth state.
引文
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[19]朱培蕾,焦仕林,姜朴,等.六价铬还原菌Cr4-1的鉴定和还原影响因素的优化[J].卫生研究,2015,44(2):201-205,210.
[20]朱玲玲,曹佳妮,张文,等.一株耐铬细菌的鉴定及其还原铬性能分析[J].环境科学学报,2013,33(10):2717-2723.
[21]何元,董兰岚,周思敏,等. 1耐铬(Ⅵ)沙雷氏菌S2的筛选及铬(Ⅵ)还原特性研究[J].现代预防医学,2017,44(18):3374-3378.
[22]林永华.六价铬还原菌Pseudomonas mandelii CH1的分离和鉴定[J].化学工程与装备,2018(9):312-316.
[2] KOTAS J,STASICKA Z. Chromium occurrence in the environment and methods of its speciation[J]. Environmental Pollution,2000,107(3):263-83.
[3]苏凤,李丽,李韵文,等.电动-还原技术修复受铬污染西部黄土的研究[J].甘肃农业大学学报,2017,52(3):90-94.
[4] BANERJEEA S,MISRA A,CHAUDHURY S,et al. A bacillus strain TCL isolated from Jharia coalmine with remarkable stress responses,chromium reduction capability and bioremediation potential[J]. Journal of Hazardous Materials,2018,367:215-223.
[5]赵光辉,常文越,李雄勇,等.铬(Ⅵ)土著还原菌筛选及初步鉴定实验研究[J].环境保护科学,2013,39(3):55-58.
[6]李维宏,杨宁,魏晓峰,等.一株Cr(Ⅵ)还原菌的筛选鉴定及其还原特性研究[J].农业环境科学学报,2015,34(11):2133-2139.
[7]冯淏.铬耐受/还原菌的多样性调研[D].杭州:浙江大学,2016.
[8]秦利玲,王天贵.用细菌解毒水溶液中六价铬的实验研究[J].化学工程师,2010,24(10):37-41.
[9] RAHMAN Z,SINGH V P. Cr(Ⅵ)reduction by Enterobacter sp. DU17 isolated from the tannery waste dump site and characterization of the bacterium and the Cr(Ⅵ)reductase[J]. International Biodeterioration&Biodegradation,2014,91:97-103.
[10]范琴,王海燕,颜湘华,等.某典型铬盐厂污染场地Cr(Ⅵ)还原菌的筛选、鉴定及还原特性[J].环境工程学报,2018,12(3):863-875.
[11]洪霞,张馨荃,严君华,等. Pseudomonas S2-3菌株对Cr(Ⅵ)的耐受性及去除[J].环境工程学报,2016,10(3):1539-1545.
[12]吴坤,张世敏.微生物学实验技术[M].北京:气象出版社,2004.
[13]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.
[14]国家环境保护总局.水和废水监测分析方法[M].北京:中国环境科学出版社,2002.
[15] FAN J J,OKYAY T O,RODRIGUES D F. The synergism of temperature,p H and growth phases on heavy metal biosorption by two environmental isolates[J]. Journal of hazardous materials. 2014,279(8):236-243.
[16]肖文丹,叶雪珠,孙彩霞,等.铬耐性菌对土壤中六价铬的还原作用[J].中国环境科学,2017,37(3):1120-1129.
[17]马小珍,费保进,金楠,等.脱硫弧菌SRB7对重金属铬Cr(Ⅵ)的还原特性[J].微生物学通报,2009,36(9):1324-1328.
[18]魏斐,杨丽荣,薛保国,等.还原六价铬细菌及其还原酶的研究[J].中国生物工程杂志,2012,32(4):53-59.
[19]朱培蕾,焦仕林,姜朴,等.六价铬还原菌Cr4-1的鉴定和还原影响因素的优化[J].卫生研究,2015,44(2):201-205,210.
[20]朱玲玲,曹佳妮,张文,等.一株耐铬细菌的鉴定及其还原铬性能分析[J].环境科学学报,2013,33(10):2717-2723.
[21]何元,董兰岚,周思敏,等. 1耐铬(Ⅵ)沙雷氏菌S2的筛选及铬(Ⅵ)还原特性研究[J].现代预防医学,2017,44(18):3374-3378.
[22]林永华.六价铬还原菌Pseudomonas mandelii CH1的分离和鉴定[J].化学工程与装备,2018(9):312-316.