Estrogen contamination is recognized as one of the most serious environmental problems, causing widespread concern worldwide. Environmental estrogens are mainly derived from human and vertebrate excretion, drugs, and agricultural activities. The use of microorganisms is currently the most economical and effective method for biodegradation of environmental estrogens. Rhodococcus equi DSSKP-R-001 (R-001) has strong estrogen-degrading capabilities. Our study indicated that R-001 can use different types of estrogen as its sole carbon source for growth and metabolism, with final degradation rates above 90%. Transcriptome analysis showed that 720 (E1), 983 (E2), and 845 (EE2) genes were significantly upregulated in the estrogen-treated group compared with the control group, and 270 differentially expressed genes (DEGs) were upregulated across all treatment groups. These DEGs included ABC transporters; estrogen-degrading genes, including those that perform initial oxidation and dehydrogenation reactions and those that further degrade the resulting substrates into small molecules; and metabolism genes that complete the intracellular transformation and utilization of estrogen metabolites through biological processes such as amino acid metabolism, lipid metabolism, carbohydrate metabolism, and the tricarboxylic acid cycle. In summary, the biodegradation of estrogens is coordinated by a metabolic network of estrogen-degrading enzymes, transporters, metabolic enzymes, and other coenzymes. In this study, the metabolic mechanisms by which Rhodococcus equi R-001 degrades various estrogens were analyzed for the first time. A new pollutant metabolism system is outlined, providing a starting point for the construction of engineered estrogen-degrading bacteria.

We screened bacteria that use E2 as its sole source of carbon and energy for growth and identified them as Rhodococcus, and we named them DSSKP-R-001. For a better understanding of the metabolic potential of the strain, whole genome sequencing of Rhodococcus DSSKP-R-001 and annotation of the functional genes were performed. The genomic sketches included a predicted protein-coding gene of approximately 5.4 Mbp with G + C content of 68.72% and 5180. The genome of Rhodococcus strain DSSKP-R-001 consists of three replicons: one chromosome and two plasmids of 5.2, 0.09, and 0.09, respectively. The results showed that there were ten steroid-degrading enzymes distributed in the whole genome of the strain. The existence and expression of estradiol-degrading enzymes were verified by PCR and RTPCR. Finally, comparative genomics was used to compare multiple strains of Rhodococcus. It was found that Rhodococcus DSSKP-R-001 had the highest similarity to Rhodococcus sp. P14 and there were 2070 core genes shared with Rhodococcus sp. P14, Rhodococcus jostii RHA1, Rhodococcus opacus B4, and Rhodococcus equi 103S, showing evolutionary homology. In summary, this study provides a comprehensive understanding of the role of Rhodococcus DSSKP-R-001 in estradiol-efficient degradation of these assays for Rhodococcus. DSSKP-R-001 in bioremediation and evolution within Rhodococcus has important meaning.

In this study, we isolated a new estrogen-degrading bacterium from a soil sample collected near a pharmaceutical factory in Beijing, China. Morphological observations, physiological and biochemical analyses, and sequence analysis showed that the strain was in the genus Acinetobacter, and it was named DSSKY-A-001. The estrogen degradation rate and growth density of strain DSSKY-A-001 were determined by high-performance liquid chromatography and a growth assay using a microplate reader, respectively. The estrogen degradation rate was 76% on the third day and 90% on the sixth day of culture. Three kinds of estrogen metabolism intermediates were detected by high-performance liquid chromatography and mass spectrometry, and the estrogen metabolic pathway and possible estrogen-degrading enzymes were predicted. RT-PCR was used to verify whether the three putative enzymes, catechol 1,2-dioxygenase, dioxygenase, and 7α-hydroxysteroid dehydrogenase, were expressed in the strain. The results of the validation were consistent with the predictions that these three enzymes were present and expressed in Acinetobacter DSSKY-A-001. To further understand the estrogen-degrading activity of the strain at the genetic level, we sequenced the genome and performed a functional gene annotation. Through this gene sequence analysis, we identified genes predicted to encode the previously detected enzymes, catechol 1,2-dioxygenase, dioxygenase, and 7α-hydroxysteroid dehydrogenase, as well as six other enzymes that may be involved in estrogen degradation. Therefore, a total of nine enzymes related to estrogen degradation were found.

In this study, we investigated environmental oestrogen 17b-estradiol (E2) degradation by a mixed culture of Rhodococcus equi DSSKP-R-001(RS) and Comamonas testosteroni QYY20150409(CT). The results demonstrated that the two strains exhibited synergistic E2 degradation activity, showing that multispecies bacterial systems can improve microbial survival and activity. The E2 degradation rate for the mixed culture system was 94% under conditions in which E2 was the sole source of carbon and growth energy, while the degradation rates of the R. equi DSSKP-R-001 and C. testosteroni QYY20150409 single strains were 86% and 76%, respectively. Comparing and analyzing the degradation products of E2, intermediate cross-metabolism products were observed in the culture containing the two bacteria. The metabolic products of benzene were essentially removed during the process of E2 degradation in the mixed culture, which showed stronger degradation ability than cultures of the individual strains. The results from this investigation on the degradation of E2 by individual and mixed bacterial cultures provide a theoretical basis for the microbial remediation of environmental oestrogens.

At present, the oestrogens detected in the environment have been classified as group 1 carcinogens by the World Health Organization. They have obvious effects on organisms at extremely low environmental concentration (1.0 ng/L) and easily accumulate in the ecosystem, which has adverse effects on the environment and ecological health. The use of microbial degradation to remove steroid estrogens from polluted environments has received increasing attention. In this study, a bacterium capable of degrading 17b-estradiol was isolated from a sewage treatment plant in Jilin, China, and identified as Serratia nematodiphila DH-S01. The results of degradation experiments showed that after culturing the bacteria for 4 days, the degradation rate of oestrone and 17b-estradiol at 15 mg/L reached 93.47% and 93.2%, respectively. Genome-wide sequencing showed that the genome of strain DH-S01 consists of a single circular chromosome, 5,256,558 bp in length, which contains 4,874 predicted coding genes. Based on genome annotation, high abundance genes are related to the metabolism of terpenoids and polyketides. Nine types of sterol-and oestrogen-degrading enzymes were annotated in this strain, and the existence and expression of the enzymes were analyzed by polymerase chain reaction (PCR) and reverse transcription polymerase chain reaction (RT-PCR). Comparative genomic analysis showed that there are genes encoding eight enzymes in the common genes of the four Serratia strains, highlighting the potential of the other three Serratia strains to degrade steroid estrogen.