Journal
ENVIRONMENTAL MICROBIOLOGY
Volume 23, Issue 11, Pages 6965-6980Publisher
WILEY
DOI: 10.1111/1462-2920.15751
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Funding
- National Key R&D Program of China [2019YFA0905500, 2016YFD0501409]
- National Natural Science Foundation of China (NSFC) [31861133002, 91951101, 31870103]
- Chinese Academy of Science [KFZD-SW-219-3, KFZD-SW-309]
- CAS Key Technology Talent Program
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Heterotrophic nitrifier strain HO-1 was identified as a novel bacterium capable of oxidizing ammonia to hydroxylamine and ultimately producing N-2 gas. Genome analysis revealed a unique denitrification pathway and the absence of genes for known ammonia monooxygenases or hydroxylamine oxidoreductases. Further experiments showed that strain HO-1 denitrified nitrite to N-2 and N2O under anaerobic and aerobic conditions, indicating a novel direct ammonia oxidation pathway named Dirammox.
Heterotrophic nitrifiers are able to oxidize and remove ammonia from nitrogen-rich wastewaters but the genetic elements of heterotrophic ammonia oxidation are poorly understood. Here, we isolated and identified a novel heterotrophic nitrifier, Alcaligenes ammonioxydans sp. nov. strain HO-1, oxidizing ammonia to hydroxylamine and ending in the production of N-2 gas. Genome analysis revealed that strain HO-1 encoded a complete denitrification pathway but lacks any genes coding for homologous to known ammonia monooxygenases or hydroxylamine oxidoreductases. Our results demonstrated strain HO-1 denitrified nitrite (not nitrate) to N-2 and N2O at anaerobic and aerobic conditions respectively. Further experiments demonstrated that inhibition of aerobic denitrification did not stop ammonia oxidation and N-2 production. A gene cluster (dnfT1RT2ABCD) was cloned from strain HO-1 and enabled E. coli accumulated hydroxylamine. Sub-cloning showed that genetic cluster dnfAB or dnfABC already enabled E. coli cells to produce hydroxylamine and further to N-15(2) from ((NH4)-N-15)(2)SO4. Transcriptome analysis revealed these three genes dnfA, dnfB and dnfC were significantly upregulated in response to ammonia stimulation. Taken together, we concluded that strain HO-1 has a novel dnf genetic cluster for ammonia oxidation and this dnf genetic cluster encoded a previously unknown pathway of direct ammonia oxidation (Dirammox) to N-2.
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