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1887

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Volume 156, Issue 7

Production of hydrogen sulfide by two enzymes associated with biosynthesis of homocysteine and lanthionine in subsp. ATCC 25586 Free

Abstract

produces a large amount of the toxic metabolite hydrogen sulfide in the oral cavity. Here, we report the molecular basis of HS production, which is associated with two different enzymes: the previously reported Cdl (Fn1220) and the newly identified Lcd (Fn0625). SDS-PAGE analysis with activity staining revealed that crude enzyme extracts from ATCC 25586 contained three major HS-producing proteins. Two of the proteins with low molecular masses migrated similarly to purified Fn0625 and Fn1220. Their kinetic values suggested that Fn0625 had a lower enzymic capacity to produce HS from -cysteine (∼30 %) than Fn1220. The Fn0625 protein degraded a variety of substrates containing C–S linkages to produce ammonia, pyruvate and sulfur-containing products. Unlike Fn0625, Fn1220 produced neither pyruvate nor ammonia from -cysteine. Reversed-phase HPLC separation and mass spectrometry showed that incubation of -cysteine with Fn1220 produced HS and an uncommon amino acid, lanthionine, which is a natural constituent of the peptidoglycans of ATCC 25586. In contrast, most of the sulfur-containing substrates tested, except -cysteine, were not used by Fn1220. Real-time PCR analysis demonstrated that the gene showed several-fold higher expression than and housekeeping genes in exponential-phase cultures of . Thus, we conclude that Fn0625 and Fn1220 produce HS in distinct manners: Fn0625 carries out -elimination of -cysteine to produce HS, pyruvate and ammonia, whereas Fn1220 catalyses the -replacement of -cysteine to produce HS and lanthionine, the latter of which may be used for peptidoglycan formation in .

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Keyword(s): PLP, pyridoxal 5′-phosphate
SGM
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2010-07-01
2025-04-29
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/content/journal/micro/10.1099/mic.0.039180-0
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Production of hydrogen sulfide by two enzymes associated with biosynthesis of homocysteine and lanthionine in Fusobacterium nucleatum subsp. nucleatum ATCC 25586
Microbiology 156, 2260 (2010); https://doi.org/10.1099/mic.0.039180-0
/content/journal/micro/10.1099/mic.0.039180-0
/content/journal/micro/10.1099/mic.0.039180-0
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