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邻苯二甲酸二正丁酯的降解菌 Rhodococcus sp. JDC-11 及其 3,4-邻苯二甲酸双加氧酶基因的分子检测

Biodegradation of di-n-butyl phthalate by Rhodococcus sp. JDC-11 and molecular detection of 3, 4-phthalate dioxygenase gene.

机构信息

Department of Bioengineering, School of Minerals Processing and Bioengineering, Central South University, Changsha 410083, China.

出版信息

J Microbiol Biotechnol. 2010 Oct;20(10):1440-5. doi: 10.4014/jmb.1004.04034.

Abstract

Rhodococcus sp. JDC-11, capable of utilizing di-n-butyl phthalate (DBP) as the sole source of carbon and energy, was isolated from sewage sludge and confirmed mainly based on 16S rRNA gene sequence analysis. The optimum pH, temperature, and agitation rate for DBP degradation by Rhodococcus sp. JDC-11 was 8.0, 30 degrees C, and 175 rpm, respectively. In addition, the effect of glucose concentration on DBP degradation indicated that low concentration of glucose inhibited the degradation of DBP while high concentrations of glucose increased its degradation. Meanwhile, the substrates utilization test showed that JDC-11 could also utilize other phthalates. Furthermore, the major metabolites of DBP degradation were identified as mono-butyl phthalate and phthalic acid by gas chromatography-mass spectrometry and the metabolic pathway of DBP degradation by Rhodococcus sp. JDC-11 was tentatively speculated. Using a set of new degenerate primer, partial sequence of the 3, 4-phthalate dioxygenase gene was obtained from the strain. Sequence analysis revealed that the phthalate dioxygenase gene of JDC-11 was highly homologous to the large subunit of phthalate dioxygenase from Rhodococcus coprophilus strain G9.

摘要

从污水污泥中分离到一株能够以邻苯二甲酸二丁酯(DBP)作为唯一碳源和能源的罗得西亚放线菌(Rhodococcus sp.)JDC-11,主要基于 16S rRNA 基因序列分析进行鉴定。罗得西亚放线菌(Rhodococcus sp.)JDC-11 降解 DBP 的最适 pH、温度和搅拌速度分别为 8.0、30°C 和 175rpm。此外,葡萄糖浓度对 DBP 降解的影响表明,低浓度的葡萄糖抑制 DBP 的降解,而高浓度的葡萄糖则增加其降解。同时,底物利用试验表明,JDC-11 还可以利用其他邻苯二甲酸酯。此外,通过气相色谱-质谱联用技术鉴定了 DBP 降解的主要代谢产物为单丁基邻苯二甲酸酯和邻苯二甲酸,并初步推测了罗得西亚放线菌(Rhodococcus sp.)JDC-11 降解 DBP 的代谢途径。使用一组新的简并引物,从该菌株中获得了邻苯二甲酸 3,4-双加氧酶基因的部分序列。序列分析表明,JDC-11 的邻苯二甲酸双加氧酶基因与 Rhodococcus coprophilus 菌株 G9 的邻苯二甲酸双加氧酶大亚基高度同源。

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