The Alcaligenes eutrophus membrane-bound hydrogenase gene locus encodes functions involved in maturation and electron transport coupling

被引:87
作者
Bernhard, M
Schwartz, E
Rietdorf, J
Friedrich, B
机构
[1] HUMBOLDT UNIV BERLIN,INST BIOL,D-10115 BERLIN,GERMANY
[2] FREE UNIV BERLIN,INST PFLANZENPHYSIOL & MIKROBIOL,D-1000 BERLIN,GERMANY
关键词
D O I
10.1128/jb.178.15.4522-4529.1996
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Alcaligenes eutrophus H16 produces two [NiFe] hydrogenases which catalyze the oxidation of hydrogen and enable the organism to utilize H-2 as the sole energy source, The genes (hoxK and hoxG) for the heterodimeric, membrane-bound hydrogenase (MBH) are located adjacent to a series of eight accessory genes (hoxZ, hoxM, hoxL, hoxO, hoxQ, hoxR, hoxT, and hoxV). In the present study, we generated a set of isogenic mutants with in-frame deletions in the two structural genes and in each of the eight accessory genes. The resulting mutants can be grouped into two classes on the basis of the H-2-oxidizing activity of the MEW. Class I mutants (hoxK Delta, hoxG Delta, hoxM Delta, hoxO Delta, and hoxQ Delta) were totally devoid of MBH-mediated, H-2-oxidizing activity. The hoxM deletion strain was the only mutant in our collection which was completely blocked in carboxy-terminal processing of large subunit HoxG, indicating that hoxM encodes a specific protease, Class II mutants (hoxZ Delta, hoxL Delta, hoxR Delta, hoxT Delta, and hoxV Delta) contained residual amounts of MBH activity in the membrane fraction of the extracts, Immunochemical analysis and Ni-63 incorporation experiments revealed that the mutations affect various steps in MBH maturation, A lesion in hoxZ led to the production of a soluble MBH which was highly active with redox dye.
引用
收藏
页码:4522 / 4529
页数:8
相关论文
共 46 条
  • [21] CLONING AND SEQUENCING OF THE GENES ENCODING THE LARGE AND THE SMALL SUBUNITS OF THE H-2 UPTAKE HYDROGENASE (HUP) OF RHODOBACTER-CAPSULATUS
    LECLERC, M
    COLBEAU, A
    CAUVIN, B
    VIGNAIS, PM
    [J]. MOLECULAR & GENERAL GENETICS, 1988, 214 (01): : 97 - 107
  • [22] THE ALCALIGENES-EUTROPHUS H16 HOXX GENE PARTICIPATES IN HYDROGENASE REGULATION
    LENZ, O
    SCHWARTZ, E
    DERNEDDE, J
    EITINGER, M
    FRIEDRICH, B
    [J]. JOURNAL OF BACTERIOLOGY, 1994, 176 (14) : 4385 - 4393
  • [23] CLONING, SEQUENCING AND CHARACTERIZATION OF THE [NIFE]HYDROGENASE-ENCODING STRUCTURAL GENES (HOXK AND HOXG) FROM AZOTOBACTER-VINELANDII
    MENON, AL
    STULTS, LW
    ROBSON, RL
    MORTENSON, LE
    [J]. GENE, 1990, 96 (01) : 67 - 74
  • [24] NUCLEOTIDE-SEQUENCES AND GENETIC-ANALYSIS OF HYDROGEN OXIDATION (HOX) GENES IN AZOTOBACTER-VINELANDII
    MENON, AL
    MORTENSON, LE
    ROBSON, RL
    [J]. JOURNAL OF BACTERIOLOGY, 1992, 174 (14) : 4549 - 4557
  • [25] MUTATIONAL ANALYSIS AND CHARACTERIZATION OF THE ESCHERICHIA-COLI HYA OPERON, WHICH ENCODES [NIFE] HYDROGENASE-1
    MENON, NK
    ROBBINS, J
    WENDT, JC
    SHANMUGAM, KT
    PRZYBYLA, AE
    [J]. JOURNAL OF BACTERIOLOGY, 1991, 173 (15) : 4851 - 4861
  • [26] CLONING, SEQUENCING, AND MUTATIONAL ANALYSIS OF THE HYB-OPERON ENCODING ESCHERICHIA-COLI HYDROGENASE-2
    MENON, NK
    CHATELUS, CY
    DERVARTANIAN, M
    WENDT, JC
    SHANMUGAM, KT
    PECK, HD
    PRZYBYLA, AE
    [J]. JOURNAL OF BACTERIOLOGY, 1994, 176 (14) : 4416 - 4423
  • [27] CLONING AND SEQUENCING OF A PUTATIVE ESCHERICHIA-COLI [NIFE] HYDROGENASE-1 OPERON CONTAINING 6 OPEN READING FRAMES
    MENON, NK
    ROBBINS, J
    PECK, HD
    CHATELUS, CY
    CHOI, ES
    PRZYBYLA, AE
    [J]. JOURNAL OF BACTERIOLOGY, 1990, 172 (04) : 1969 - 1977
  • [28] Miller J.H., 1972, ASSAY BETA GALACTOSI
  • [29] SITE-DIRECTED MUTAGENESIS OF THE HYDROGENASE SIGNAL PEPTIDE CONSENSUS BOX PREVENTS EXPORT OF A BETA-LACTAMASE FUSION PROTEIN
    NIVIERE, V
    WONG, SL
    VOORDOUW, G
    [J]. JOURNAL OF GENERAL MICROBIOLOGY, 1992, 138 : 2173 - 2183
  • [30] NUCLEOTIDE-SEQUENCE AND ORGANIZATION OF AN H2-UPTAKE GENE-CLUSTER FROM RHIZOBIUM-LEGUMINOSARUM BV VICIAE CONTAINING A RUBREDOXIN-LIKE GENE AND 4 ADDITIONAL OPEN READING FRAMES
    REY, L
    HIDALGO, E
    PALACIOS, J
    RUIZARGUESO, T
    [J]. JOURNAL OF MOLECULAR BIOLOGY, 1992, 228 (03) : 998 - 1002