Inhibition of Intracellular Succinate Dehydrogenase of the Bacillus subtilis Bacterium as a Criterion for the Involvement of an Enzyme in the Reduction of Chloride Iodonitrotetrazolium, an Indicator of Cell Viability

Mammalian and Escherichia coli succinate dehydrogenase (SDH) and E. coli fumarate reductase apparently contain an essential cysteine residue at the active site, as shown by substrate-protectable inactivation with thiol-specific reagents. Bacillus subtilis SDH was found to be resistant to this type of reagent and contains an alanine residue at the amino acid position equivalent to the only invariant cysteine in the flavoprotein subunit of E. coli succinate oxidoreductases. Substitution of this alanine, at position 252 in the flavoprotein subunit of B. subtilis SDH, by cysteine resulted in an enzyme sensitive to thiol-specific reagents and protectable by substrate. Other biochemical properties of the redesigned SDH were similar to those of the wild-type enzyme. It is concluded that the invariant cysteine in the flavoprotein of E. coli succinate oxidoreductases corresponds to the active site thiol. However, this cysteine is most likely not essential for succinate oxidation and seemingly lacks an assignable specific function. An invariant arginine in juxtaposition to Ala-252 in the flavoprotein of B. subtilis SDH, and to the invariant cysteine in the E. coli homologous enzymes, is probably essential for substrate binding.

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Characterization of cspB, a Bacillus subtilis inducible cold shock gene affecting cell viability at low temperatures.

Journal of Bacteriology ◽

10.1128/jb.174.20.6326-6335.1992 ◽

1992 ◽

Vol 174 (20) ◽

pp. 6326-6335 ◽

Cited By ~ 146

Author(s):

G Willimsky ◽

H Bang ◽

G Fischer ◽

M A Marahiel

Keyword(s):

Bacillus Subtilis ◽

Cell Viability ◽

Low Temperatures ◽

Cold Shock ◽

Shock Gene

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Gene replacement method for determining conditions in which Bacillus subtilis genes are essential or dispensable for cell viability

Applied Microbiology and Biotechnology ◽

10.1007/s00253-003-1502-5 ◽

2004 ◽

Vol 64 (3) ◽

pp. 382-386 ◽

Cited By ~ 8

Author(s):

H. Yakhnin ◽

P. Babitzke

Keyword(s):

Bacillus Subtilis ◽

Cell Viability ◽

Gene Replacement ◽

Replacement Method

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Effects of Bacillus Subtilis-Fermented White Sword Bean Extract on Adipogenesis and Lipolysis of 3T3-L1 Adipocytes

Foods ◽

10.3390/foods10061423 ◽

2021 ◽

Vol 10 (6) ◽

pp. 1423

Author(s):

Yujeong Choi ◽

Da-Som Kim ◽

Min-Chul Lee ◽

Seulgi Park ◽

Joo-Won Lee ◽

...

Keyword(s):

Bacillus Subtilis ◽

Cell Viability ◽

Dietary Supplement ◽

Free Glycerol ◽

Glycerol Content ◽

Treatment Groups ◽

Protein Levels ◽

Triglyceride Accumulation ◽

Before And After

To investigate the adipogenesis and lipolysis effects of the Bacillus subtilis-fermented white sword bean extract (FWSBE) on 3T3-L1 adipocytes, we treated 3T3-L1 preadipocytes before and after differentiation with FWSBE and measured triglyceride, free glycerol, mRNA, and protein levels. First, FWSBE reduced the cell viability of 3T3-L1 pre-adipocytes under 1000 µg/mL conditions. Triglyceride accumulation in 3T3-L1 pre-adipocytes was suppressed, and free glycerol content in mature 3T3-L1 adipocytes was increased in the FWSBE treatment groups, indicating that FWSBE has anti-obesity effects. Further, FWSBE suppressed adipogenesis in 3T3-L1 pre-adipocytes by lowering the protein levels of C/EBPα, PPARγ, and FAS and increasing the level of pACC and pAMPK. Additionally, FWSBE promoted lipolysis in mature 3T3-L1 adipocytes by increasing the transcription levels of Ppara, Acox, and Lcad and the protein levels of pHSL and ATGL. Thus, we suggest that FWSBE can be a potential dietary supplement because of its anti-obesity properties.

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