1887

Abstract

produces an abundance of hydrogen sulfide (HS) in the oral cavity that is mediated by several enzymes. The identification and characterization of three distinct enzymes (Fn0625, Fn1055 and Fn1220) in that catalyse the production of HS from -cysteine have been reported. In the current study, a novel enzyme involved in the production of HS in ATCC 25586, whose molecular mass had been estimated to be approximately 130 kDa, was identified by two-dimensional electrophoresis combined with MALDI-TOF MS. The enzyme, Fn1419, has previously been characterized as an -methionine γ-lyase. SDS-PAGE and gel-filtration chromatography indicated that Fn1419 has a molecular mass of 43 kDa and forms tetramers in solution. Unlike other enzymes associated with HS production in , the quaternary structure of Fn1419 was not completely disrupted by exposure to SDS. The purified recombinant enzyme exhibited a of 0.32±0.02 mM and a of 0.69±0.01 s. Based on current and published data, the enzymic activity for HS production from -cysteine in is ranked as follows: Fn1220>Fn1055>Fn1419>Fn0625. Based on kinetic values and relative mRNA levels of the respective genes, as determined by real-time quantitative PCR, the amount of HS produced by Fn1419 was estimated to be 1.9 % of the total HS produced from -cysteine in ATCC 25586. In comparison, Fn1220 appeared to contribute significantly to HS production (87.6 %).

Funding
This study was supported by the:
  • , Ministry of Education, Culture, Sports, Science and Technology, Japan , (Award 23592721)
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2011-10-01
2021-01-26
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