Fe(III)-Aided Novosphingobium sp. ES2-1 Regulates Molecular Mechanisms of 17β-Estradiol Biodegradation

17β-estradiol (E2) is one of the strongest environmental estrogens threatening wildlife and human health globally. Microbial degradation is an alternative strategy to remediate E2-contaminated sites and may be regulated by ubiquitous Fe(III) in eco-environments. We have previously obtained a high-efficiency E2 degrader, Novosphingobium sp. ES2-1, and investigated its metabolic pathway in connection with monooxygenase EstO1-induced ring-B opening; however, the molecular mechanisms of ring-A cleavage in E2 are sorely lacking, especially under Fe(III)-aided regulation. Here, an extradiol dioxygenase EstN1 from strain ES2-1 involved in the ring-A cleavage of E2 was reported. It catalyzed the 4,5-seco reaction of 4-hydroxyestrone (4-OH-E1, a key E2-oxidized intermediate) with the support of the electron transport chain consisting of ferredoxin EstN2 and ferredoxin reductase EstN3, resulting in a ring-A meta-cleaved product. Interestingly, Fe(III)-assisted strain ES2-1 consolidated the opening of rings A and B in E2 by reinforcing the expression of estO1 and estN1 genes, consequently enhancing E2 metabolism. Compared to Fe(III) starvation, the biodegradation half-life of E2 was sharply reduced from 1.35 to 0.59 d after Fe(III) supplementation. Simultaneously, the transcription of estO1 and estN1 genes increased clearly from 4.3 to 47.5 times and 6.6 to 246.8 times after Fe(III) induction, respectively, accompanied by remarkable improvement in the abundance of ring-A/B cleavage products and their pyridine derivatives. These findings highlight the significance of Fe(III) in regulating the microbial remediation of environmental estrogens at the molecular level.