Isolation of a unique benzothiophene-desulphurizing bacterium, Gordona sp. strain 213E (NCIMB 40816), and characterization of the desulphurization pathway
Gordona sp. strain 213E (NCIMB 40816) grew in pure culture in a mineral salts medium containing fructose as a source of carbon and energy, and benzothiophene (BTH) as the sole source of sulphur. During growth a phenolic compound accumulated, as indicated by the production of a blue colour on addition of Gibb's reagent. Therefore this pathway is analogous to the dibenzothiophene (DBT) desulphurization pathway of Rhodococcus sp. strain IGTS8, in which 2-hydroxybiphenyl accumulates during growth with DBT as the sole sulphur source. Ethyl acetate extraction of the culture medium yielded the metabolites benzothiophene S-oxide (BTHO), benzothiophene S,S-dioxide (BTHO2), benzo[c][1,2]oxathiin 6-oxide (BcOTO), 2-(2'-hydroxyphenyl)ethan 1-al (HPEal) and benzofuran (BFU). The deduced pathway for BTH desulphurization is BTH ? BTHO ? BTHO2 ? HPESi- ? HPEal. HPESi- is (Z)-2-(2'-hydroxyphenyl)ethen 1-sulphinate, the stable aqueous-solution form of BcOTO. It was concluded that HPEal was the Gibb's-reagent-reactive phenolic compound which accumulated in the culture medium of strain 213E during growth, and that the presence of BFU was due to partial condensation of HPEal during the ethyl acetate extraction procedure. Gordona sp. strain 213E was unable to grow in a mineral salts medium containing fructose as a source of carbon and energy and DBT as the sole sulphur source. BTH-desulphurization-active cells (grown using BTH as sole sulphur source) were unable to desulphurize DBT. Likewise Rhodococcus sp. strain IGTS8 was unable to grow using BTH as the sole sulphur source, and DBT-desulphurization-active cells of strain IGTS8 (grown using DBT as sole sulphur source) were unable to desulphurize BTH. This absence of cross-reactivity is discussed in terms of fundamental differences in the chemistry of the DBT- and BTH-desulphurization reactions.
AsturiasJ.A.,
EltisL.D.,
PruchaM.,
TimmisK.M.1994; Analysis of three 2,3-dihydroxybiphenyl 1,2-dioxygenases found in Rhodococcus globerulus P6.. Biol Chem 269:7807–7815
BeilS.,
KehrliH.,
JamesP.,
StaudenmannW.,
CookA.M.,
LeisingerT.,
KerteszM.A.1995; Purification and charac-terisation of the arylsulfatase synthesised by Pseudomonas aeruginosa PAO during growth in sulfate-free medium and cloning of the arylsulfatase gene (atsA).. Eur J Biochem 229:385–394
BowieJ.H.,
WilliamsD.H.,
LawessonS.-O.,
MadsenJ.ø.,
NoldeC.,
SchrollG.1966; Studies in mass spectrometry. XV. Mass spectra of sulphoxides and sulphones. The formation of C-C and C-O bonds upon electron impact.. Tetrahedron 22:3515–3525
ChiangY.,
KresgeA.J.,
WalshP.A.,
YinY.1989; Phenyl- acetaldehyde and its cis- and trans-enols and enolate ions. Determination of the cis: trans ratio under equilibrium and kinetic control.. J Chem Soc Chem Commun869–871
ConstantiM.,
GirlatJ.,
BordonsA.1996; Degradation and desulfurization of dibenzothiophene sulfone and other sulfur compounds by Agrobacterium MC501 and a mixed culture.. Enzyme Microb Technol 19:214–219
DenomeS.A.,
OlsonE.S.,
YoungK.D.1993; Identification and cloning of genes involved in specific desulphurisation of dibenzothiophene by Rhodococcus sp. strain IGTS8.. Appl Environ Microbiol 59:2837–2843
KayserK.J.,
Bielaga-JonesB.A.,
JackowskiK.,
OdusanO.,
KilbaneJ.J.1993; Utilization of organosulphur compounds by axenic and mixed cultures of Rbodococcus rbodochrous strain IGTS8.. J Gen Microbiol 139:3123–3129
KosonoS.,
MaedaM.,
FujiF.,
AraiH.,
KudoT.1997; Three of the seven bphC genes of Rbodococcus erytbropolis TA421, isolated from a termite ecosystem, are located on an indigenous plasmid associated with biphenyl degradation.. Appl Environ Microbiol 63:3283–3285
LeiB.,
TuS.-C.1996; Gene overexpression, purification and identification of a desulfurization enzyme from Rbodococcus sp. strain IGTS8 as a sulfide/sulfoxide monooxygenase.. J Bacteriol 17:5699–5705
LiM.Z.,
SquiresC.H.,
ChildsJ.D.1996; Genetic analysis of the dsz promoter and associated regulatory regions of Rbodococcus erytbropolis IGTS8.. J Bacteriol 178:6409–6418
MasaiE.,
SugiyamaK.,
IwashiteN.,
ShimizuS.,
HauschildJ.E.,
HattaT.,
KimbaraK.,
YanoK.,
FukudaM.1997; The bphDEF meta-cleavage pathway genes involved in biphenyl/ polychlorinated biphenyl degradation are located on a linear plasmid and separated from the initial bphhCB genes in Rbodococcus sp. strain RHA1.. Gene 187:141–149
OhshiroT.,
HirataT.,
IzumiY.1996a; Desulfurization of dibenzothiophene derivatives by whole cells of Rbodococcus erytbropolis strain H-2.. FEMS Microbiol Lett 142:65–70
OlsonE.S.,
StanleyD.C.,
GallagherJ.R.1993; Charac-terisation of intermediates in the microbial desulfurization of dibenzothiophene.. Energy & Fuels 7:159–164
van der PloegJ.R.,
Iwanikca-NowickaR.,
KerteszM.A.,
LeisingerT.,
HoyniewiczM.M.1998; Involvement of CysB and Cbl regulatory proteins in the tauABCD operon and other sulphate starvation inducible genes in Escherichia coli.. J Bacteriol 179:7671–7678
RheeS.-K.,
ChangJ.H.,
ChangY.K.,
ChangH.O.1998; Desulfurization of dibenzothiophene and diesel oils by a newly isolated Gordona strain, CYKS1.. Appl Environ Microbiol 64:2327–2331
WangP.,
KrawiecS.1996; Kinetic analyses of desulfurization of dibenzothiophene by Rbodococcus erytbropolis in batch and fed-batch cultures Appl Environ Microbiol
. 62:3066–3068
WangP.,
HumphreyA.E.,
KrawiecS.1996; Kinetic analyses of desulfurization of dibenzothiophene by Rbodococcus erytbropolis in continuous cultures.. Appl Environ Microbiol 62:3066–3068
Isolation of a unique benzothiophene-desulphurizing bacterium, Gordona sp. strain 213E (NCIMB 40816), and characterization of the desulphurization pathway