Denitrobacterium

The sole species currently described in the genus Denitrobacterium, D. detoxificans, is a Gram-positive, obligate anaerobe. In the study conducted by Anderson et al., all of the four strains (NPOH1, NPOH2, NPOH3, and MAJ1) are shown to possess high G+C content in their DNAs (60, 58, 56, and 60 mol%, respectively) and are closely related to one another (more than 99% sequence identity). Additionally, the closest intergeneric relative is Coriobacterium glomerans with 86% sequence identity, based on the 16S rRNA sequence comparison between the NPOH1 strain and sequences available in GenBank.

In the bovine rumen, Denitrobacterium detoxificans metabolizes the following substrates through oxidation:{{Cite journal |last1=Ochoa-García |first1=Pedro Antonio |last2=Anderson |first2=Robin C. |last3=Arévalos-Sánchez |first3=Martha María |last4=Rodríguez-Almeida |first4=Felipe Alonso |last5=Félix-Portillo |first5=Monserrath |last6=Muro-Reyes |first6=Alberto |last7=Božić |first7=Aleksandar K. |last8=Arzola-Álvarez |first8=Claudio |last9=Corral-Luna |first9=Agustín |date=2021-08-16 |title=Astragallus mollissimus plant extract: a strategy to reduce ruminal methanogenesis |url=https://doi.org/10.1007/s11250-021-02882-1 |journal=Tropical Animal Health and Production |language=en |volume=53 |issue=4 |pages=436 |doi=10.1007/s11250-021-02882-1 |pmid=34401959 |s2cid=237148677 |issn=1573-7438}}

• Hydrogen
• Formate
• Lactate

The oxidation of these above compounds are coupled with the reduction of nitrocompounds such as:

• 3-nitro-1-propionic acid (3NPA)
• 3-nitro-1-propanol (3NPOH)
• 2-nitroethane (2NEOH), an analogue of 3NPOH

There are speculations as to how these nitrocompounds are metabolized. The primary mechanism of 3NPA and 3NPOH metabolism is the reduction to amines, i.e. β-alanine and aminopropanol, respectively.{{Cite journal |last1=Anderson |first1=R C |last2=Rasmussen |first2=M A |last3=Allison |first3=M J |date=September 1993 |title=Metabolism of the plant toxins nitropropionic acid and nitropropanol by ruminal microorganisms |url=http://dx.doi.org/10.1128/aem.59.9.3056-3061.1993 |journal=Applied and Environmental Microbiology |volume=59 |issue=9 |pages=3056–3061 |doi=10.1128/aem.59.9.3056-3061.1993 |pmid=8215375 |pmc=182406 |bibcode=1993ApEnM..59.3056A |issn=0099-2240}} β-alanine is further metabolized by ruminal microorganisms, whereas aminopropanol seems to be a final product. It is also speculated that nitrite may be cleaved off from both 3NPA and 3NPOH as a minor metabolite, which is then further reduced to ammonia.{{Cite journal |last1=Majak |first1=W. |last2=Cheng |first2=K.-J. |date=1981-07-01 |title=Identification of rumen bacteria that anaerobically degrade aliphatic nitrotoxins |url=http://dx.doi.org/10.1139/m81-099 |journal=Canadian Journal of Microbiology |volume=27 |issue=7 |pages=646–650 |doi=10.1139/m81-099 |pmid=7197575 |issn=0008-4166}} 3NPA gets metabolized by ruminal microbes more rapidly than 3NPOH;{{Cite journal |last1=Gustine |first1=D. L. |last2=Moyer |first2=B. G. |last3=Wangsness |first3=P. J. |last4=Shenk |first4=J. S. |date=1977-06-01 |title=Ruminal Metabolism of 3-Nitropropanoyl-D-Glucopyranoses from Crownvetch |url=http://dx.doi.org/10.2527/jas1977.4461107x |journal=Journal of Animal Science |volume=44 |issue=6 |pages=1107–1111 |doi=10.2527/jas1977.4461107x |issn=0021-8812}}{{Cite journal |last1=MAJAK |first1=W. |last2=CLARK |first2=L. J. |title=Metabolism of Aliphatic Nitro Compounds in Bovine Rumen Fluid |date=1980-06-01 |journal=Canadian Journal of Animal Science |volume=60 |issue=2 |pages=319–325 |doi=10.4141/cjas80-041 |issn=0008-3984 |doi-access=free}} therefore, 3NPA is less toxic to ruminants grazing on leguminous plants containing the conjugates of these nitrocompounds than 3NPOH.

The above nitrocompounds are abundant in many forages in the forms of glycosides and glucose esters.{{Cite journal |last1=Anderson |first1=Robin C. |last2=Majak |first2=Walter |last3=Rassmussen |first3=Mark A. |last4=Callaway |first4=Todd R. |last5=Beier |first5=Ross C. |last6=Nisbet |first6=David J. |last7=Allison |first7=Milton J. |date=2005-03-01 |title=Toxicity and Metabolism of the Conjugates of 3-Nitropropanol and 3-Nitropropionic Acid in Forages Poisonous to Livestock |url=https://pubs.acs.org/doi/10.1021/jf040392j |journal=Journal of Agricultural and Food Chemistry |language=en |volume=53 |issue=6 |pages=2344–2350 |doi=10.1021/jf040392j |pmid=15769179 |issn=0021-8561}} Miserotoxin is the most common glycoside of 3NPOH as 3-nitro-1-propyl-β-_D-glucopyranoside, first isolated from Astragalus oblongifolius.{{Cite journal |last1=Stermitz |first1=Frank R. |last2=Norris |first2=Frank A. |last3=Williams |first3=Miles Coburn |date=July 1969 |title=Miserotoxin, new naturally occurring nitro compound |url=http://dx.doi.org/10.1021/ja01044a078 |journal=Journal of the American Chemical Society |volume=91 |issue=16 |pages=4599–4600 |doi=10.1021/ja01044a078 |issn=0002-7863}} Other glycosides of 3NPOH include β-_D-gentiobioside,{{Cite journal |last1=Majak |first1=Walter |last2=Benn |first2=Michael H. |date=1988 |title=3-Nitro-1-propyl-β-d-gentiobioside from Astragalus miser var. serotinus |url=http://dx.doi.org/10.1016/0031-9422(88)80279-6 |journal=Phytochemistry |volume=27 |issue=4 |pages=1089–1091 |doi=10.1016/0031-9422(88)80279-6 |bibcode=1988PChem..27.1089M |issn=0031-9422}} allolactoside,{{Cite journal |last1=Majak |first1=W. |last2=Benn |first2=M. H. |last3=Huang |first3=Y. Y. |date=September 1988 |title=A New Glycoside of 3-Nitropropanol from Astragalus miser var. serotinus |url=http://dx.doi.org/10.1021/np50059a032 |journal=Journal of Natural Products |volume=51 |issue=5 |pages=985–988 |doi=10.1021/np50059a032 |pmid=21401184 |issn=0163-3864}} laminaribioside,{{Cite journal |last1=Benn |first1=Michael H. |last2=Majak |first2=Walter |date=January 1989 |title=3-Nitro-1-propyl-β-d-laminaribioside from Astragalus miser var. serotinus |url=http://dx.doi.org/10.1016/s0031-9422(00)97986-x |journal=Phytochemistry |volume=28 |issue=9 |pages=2369–2371 |doi=10.1016/s0031-9422(00)97986-x |bibcode=1989PChem..28.2369B |issn=0031-9422}} and cellobioside{{Cite journal |last1=Long |first1=Melissa |last2=Benn |first2=Michael |last3=Majak |first3=Walter |last4=McDiarmid |first4=Ruth |date=January 1992 |title=3-nitropropyl glycosides of Astragalus miser var. serotinus |url=http://dx.doi.org/10.1016/0031-9422(91)83063-q |journal=Phytochemistry |volume=31 |issue=1 |pages=321–323 |doi=10.1016/0031-9422(91)83063-q |bibcode=1992PChem..31..321L |issn=0031-9422}} from Astragalus miser var. serotinus. Glucose esters of 3NPA are produced by species of the genera Coronilla{{Cite journal |last=Majak |first=W |date=1976 |title=Nitropropanylglucopyranoses in Coronilla varia |url=http://dx.doi.org/10.1016/s0031-9422(00)86835-1 |journal=Phytochemistry |volume=15 |issue=3 |pages=415–417 |doi=10.1016/s0031-9422(00)86835-1 |bibcode=1976PChem..15..415M |issn=0031-9422}}{{Cite journal |last1=Moyer |first1=Barton G. |last2=Pfeffer |first2=Philip E. |last3=Moniot |first3=Jerry L. |last4=Shamma |first4=Maurice |last5=Gustine |first5=David L. |date=January 1977 |title=Corollin, coronillin and coronarian: Three new 3-nitropropanoyl-d-glucopyranoses from Coronilla varia |url=http://dx.doi.org/10.1016/0031-9422(77)80068-x |journal=Phytochemistry |volume=16 |issue=3 |pages=375–377 |doi=10.1016/0031-9422(77)80068-x |bibcode=1977PChem..16..375M |issn=0031-9422}}{{Cite journal |last1=Majak |first1=Walter |last2=Benn |first2=Michael |date=1994 |title=Additional esters of 3-nitropropanoic acid and glucose from fruit of the New Zealand karaka tree, Corynocarpus laevigatus |url=http://dx.doi.org/10.1016/s0031-9422(00)90635-6 |journal=Phytochemistry |volume=35 |issue=4 |pages=901–903 |doi=10.1016/s0031-9422(00)90635-6 |bibcode=1994PChem..35..901M |issn=0031-9422}}, Astragalus{{Cite journal |last1=Stermitz |first1=F.R. |last2=Lowry |first2=W.T. |last3=Ubben |first3=E. |last4=Sharifi |first4=I. |date=January 1972 |title=1,6-Di-3-nitropropanoyl-β-d-glucopyranoside from Astragalus cibarius |url=http://dx.doi.org/10.1016/s0031-9422(00)89851-9 |journal=Phytochemistry |volume=11 |issue=12 |pages=3525–3527 |doi=10.1016/s0031-9422(00)89851-9 |bibcode=1972PChem..11.3525S |issn=0031-9422}}, Indigofera{{Cite journal |last1=Finnegan |first1=R.A. |last2=Stephani |first2=R.A. |date=February 1968 |title=Structure of Hiptagin as 1,2,4,6-tetra-O-(3-nitropropanoyl)-β-D-Glucopyranoside, its Identity with Endecaphyllin X, and the Synthesis of its Methyl Ether |url=http://dx.doi.org/10.1002/jps.2600570233 |journal=Journal of Pharmaceutical Sciences |volume=57 |issue=2 |pages=353–354 |doi=10.1002/jps.2600570233 |pmid=5641692 |issn=0022-3549}}{{Cite journal |last1=Benn |first1=Michael |last2=Mcewan |first2=Denise |last3=Pass |first3=Michael A. |last4=Majak |first4=Walter |date=July 1992 |title=Three nitropropanoyl esters of glucose from Indigofera linnaei |url=http://dx.doi.org/10.1016/0031-9422(92)83284-6 |journal=Phytochemistry |volume=31 |issue=7 |pages=2393–2395 |doi=10.1016/0031-9422(92)83284-6 |bibcode=1992PChem..31.2393B |issn=0031-9422}}{{Cite journal |last1=Garcez |first1=Walmir S. |last2=Garcez |first2=Fernanda R. |last3=Honda |first3=Neli K. |last4=da Silva |first4=Antonio J.R. |date=January 1989 |title=A nitropropanoyl-glucopyranoside from Indigofera suffruticosa |url=http://dx.doi.org/10.1016/0031-9422(89)80220-1 |journal=Phytochemistry |volume=28 |issue=4 |pages=1251–1252 |doi=10.1016/0031-9422(89)80220-1 |bibcode=1989PChem..28.1251G |issn=0031-9422}}, and Hiptage. 3NPA is also produced by Astragalus canadensis in the forms of oxotetrahydrofuranyl{{Cite journal |last=Benn |first=M |date=December 1995 |title=Aliphatic nitro-compounds in Astragalus canadensis |url=http://dx.doi.org/10.1016/0031-9422(95)00482-m |journal=Phytochemistry |volume=40 |issue=6 |pages=1629–1631 |doi=10.1016/0031-9422(95)00482-m |bibcode=1995PChem..40.1629B |issn=0031-9422}} and isoxazolinone esters.{{Cite journal |last1=Benn |first1=Michael H |last2=Majak |first2=Walter |last3=Aplin |first3=Robin |date=July 1997 |title=A nitropropanoyl isoxazolinone derivative in two species of Astragalus |url=http://dx.doi.org/10.1016/s0305-1978(97)00022-7 |journal=Biochemical Systematics and Ecology |volume=25 |issue=5 |pages=467–468 |doi=10.1016/s0305-1978(97)00022-7 |issn=0305-1978}}

The D. detoxifican strain NPOH1 was first isolated and cultured in the 1996 study by Anderson et al., investigating the metabolism of nitrotoxins such as 3-nitro-1-propanol and 3-nitro-1-propionate. Strains NPOH2 and NPOH3 were isolated from a roll tube containing an agar medium with energy-depleted rumen fluid (at 40% v/v), sodium carbonate, resazurin, _L-cysteine-HCl, lipoic acid, vitamins, minerals, and Amisoy (a partially purified soy protein product by Quest International, at 0.08% w/v), supplemented with 9 mM 3-nitro-1-propanol and inoculated with 2 x 10⁻⁴ mL of nonenriched ruminal fluid. The rumen contents containing NPOH1, NPOH2, and NPOH3 were obtained from two different cows (one with NPOH1 and another with NPOH2&3) at the National Animal Disease Center (NADC) in Ames, IA, USA. Strain MAJ1 was isolated from rumen contents of a cow on a milkvetch range harboring Astragalus miser var. serotinus in British Columbia, Canada.

In the 2000 article, Anderson et al. proposed the assignment of the novel bacteria into the class Actinobacteria, subclass Coriobacteridae, order Coriobacteriales, family Coriobacteriaceae based on the high mole percent G+C content and 16S rRNA sequence. The genus Denitrobacterium was included in the family Coriobaceteriaceae by Zhi et al. in the 2009 publication{{Cite journal |last1=Zhi |first1=X.-Y. |last2=Li |first2=W.-J. |last3=Stackebrandt |first3=E. |date=2009-03-01 |title=An update of the structure and 16S rRNA gene sequence-based definition of higher ranks of the class Actinobacteria, with the proposal of two new suborders and four new families and emended descriptions of the existing higher taxa |journal=International Journal of Systematic and Evolutionary Microbiology |language=en |volume=59 |issue=3 |pages=589–608 |doi=10.1099/ijs.0.65780-0 |pmid=19244447 |issn=1466-5026 |doi-access=free}} on addendum to the class Actinobacteria.

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