Two membrane anchors of Wolinella succinogenes hydrogenase and their function in fumarate and polysulfide respiration

被引:45
作者
Gross, R [1 ]
Simon, J [1 ]
Theis, F [1 ]
Kröger, A [1 ]
机构
[1] Univ Frankfurt, Inst Mikrobiol, D-60439 Frankfurt, Germany
关键词
membrane-integrated Ni-hydrogenase; cytochrome b; Wolinella succinogenes; fumarate respiration; polysulfide respiration;
D O I
10.1007/s002030050614
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Wolinella succinogenes can grow by anaerobic respiration with fumarate or polysulfide as the terminal electron acceptor, and H-2 or formate as the electron donor. A Delta hydABC mutant lacking the hydrogenase structural genes did not grow with H-2 and either fumarate or polysulfide. In contrast to the wild-type strain, the mutant grown with fumarate and with formate instead of H-2 did not catalyze the reduction of fumarate, polysulfide, dimethylnaphthoquinone, or benzyl viologen by H-2. Growth and enzymic activities were restored upon integration of a plasmid carrying hydABC into the genome of the Delta hydABC mutant. The Delta hydABC mutant was complemented with hydABC operons modified by artificial stop codons in hydA (StopA) or at the 5'-end of hydC (StopC). The StopC mutant lacked HydC, and the hydrophobic C-terminus of HydA was missing in the hydrogenase of the StopA mutant. The two mutants catalyzed benzyl viologen reduction by H-2. The enzyme activity was located in the membrane of the mutants. A mutant with both modifications (StopAC) contained the activity in the periplasm. The three mutants did not grow with H-2 and either fumarate or polysulfide, and did not catalyze dimethylnaphthoquinone reduction by H2 We conclude that the same hydrogenase serves in the anaerobic respiration with fumarate and with polysulfide. HydC and the C-terminus of HydA appear to be required for both routes of electron transport and for dimethylnaphthoquinone reduction by H2. The hydrogenase is anchored in the membrane by HydC and by the C-terminus of HydA. The catalytic subunit HydB is oriented towards the periplasmic side of the membrane.
引用
收藏
页码:50 / 58
页数:9
相关论文
共 50 条
  • [1] BASIC LOCAL ALIGNMENT SEARCH TOOL
    ALTSCHUL, SF
    GISH, W
    MILLER, W
    MYERS, EW
    LIPMAN, DJ
    [J]. JOURNAL OF MOLECULAR BIOLOGY, 1990, 215 (03) : 403 - 410
  • [2] BERGMEYER HU, 1974, METHODEN ENZYMATISCH, P689
  • [3] SEQUENCE-ANALYSIS OF SUBUNITS OF THE MEMBRANE-BOUND NITRATE REDUCTASE FROM A DENITRIFYING BACTERIUM - THE INTEGRAL MEMBRANE SUBUNIT PROVIDES A PROTOTYPE FOR THE DIHEME ELECTRON-CARRYING ARM OF A REDOX LOOP
    BERKS, BC
    PAGE, MD
    RICHARDSON, DJ
    REILLY, A
    CAVILL, A
    OUTEN, F
    FERGUSON, SJ
    [J]. MOLECULAR MICROBIOLOGY, 1995, 15 (02) : 319 - 331
  • [4] Functional and structural role of the cytochrome b subunit of the membrane-bound hydrogenase complex of Alcaligenes eutrophus H16
    Bernhard, M
    Benelli, B
    Hochkoeppler, A
    Zannoni, D
    Friedrich, B
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 1997, 248 (01): : 179 - 186
  • [5] BODE C, 1968, Z KLIN CHEM KLIN BIO, V6, P418
  • [6] CLONING AND NUCLEOTIDE-SEQUENCE OF THE STRUCTURAL GENES ENCODING THE FORMATE DEHYDROGENASE OF WOLINELLA-SUCCINOGENES
    BOKRANZ, M
    GUTMANN, M
    KORTNER, C
    KOJRO, E
    FAHRENHOLZ, F
    LAUTERBACH, F
    KROGER, A
    [J]. ARCHIVES OF MICROBIOLOGY, 1991, 156 (02) : 119 - 128
  • [7] CONSTRUCTION AND CHARACTERIZATION OF NEW CLONING VEHICLES .2. MULTIPURPOSE CLONING SYSTEM
    BOLIVAR, F
    RODRIGUEZ, RL
    GREENE, PJ
    BETLACH, MC
    HEYNEKER, HL
    BOYER, HW
    CROSA, JH
    FALKOW, S
    [J]. GENE, 1977, 2 (02) : 95 - 113
  • [8] BIOSYNTHETIC PATHWAYS OF VIBRIO-SUCCINOGENES GROWING WITH FUMARATE AS TERMINAL ELECTRON-ACCEPTOR AND SOLE CARBON SOURCE
    BRONDER, M
    MELL, H
    STUPPERICH, E
    KROGER, A
    [J]. ARCHIVES OF MICROBIOLOGY, 1982, 131 (03) : 216 - 223
  • [9] HIGH-EFFICIENCY TRANSFORMATION OF ESCHERICHIA-COLI BY HIGH-VOLTAGE ELECTROPORATION
    DOWER, WJ
    MILLER, JF
    RAGSDALE, CW
    [J]. NUCLEIC ACIDS RESEARCH, 1988, 16 (13) : 6127 - 6145
  • [10] VALUATED MATROIDS
    DRESS, AWM
    WENZEL, W
    [J]. ADVANCES IN MATHEMATICS, 1992, 93 (02) : 214 - 250