International Journal of Hydrogen Energy 31 (2006) 531 – 538

www.elsevier.com/locate/ijhydene

Effect of changes in the level of light harvesting complexes of

Rhodobacter sphaeroides on the photoheterotrophic production
of hydrogen
Eui-Jin Kima , Ju-Sim Kima,1 , Mi-Sun Kimb , Jeong K. Leea,∗
a Department of Life Science and Interdisciplinary Program of Integrated Biotechnology, Sogang University, Mapo, Shinsu 1,
Seoul 121-742, Republic of Korea
b Biomass Research Team, Korea Institute of Energy Research, Daejeon 305-343, Republic of Korea

Received 29 March 2005; received in revised form 4 April 2005; accepted 14 April 2005
Available online 14 June 2005

Abstract
Increase in levels of photosynthetic spectral complexes by maintaining the plasmids harboring DNA encoding puhA, pufBA,
or pucBAC in trans in Rhodobacter sphaeroides resulted in decrease of the photoheterotrophic production of H2 . However,
removal of B875 or B800–850 light-harvesting (LH) complexes affected H2 production differently. Lack of B875 complex
following in-frame deletion of pufBA (mutant PUF1) not only slowed photoheterotrophic growth but also decreased H2
production, indicative of the essential requirement of the complex for LH process. However, the pucBA-deleted mutant, PUC1
lacking of B800–850 complex, increased H2 production in comparison with its parental cell by approximately twofold, given
irradiated with light (10 W/m2 ) saturating the growth of wild type. The H2 production of PUC1 did not increase in proportion
to the light intensity, which is also observed with wild type. Thus, we suggest that light is not limited for the H2 production
of the cells under the experimental conditions employed in this work, but the cellular energy to be used for the formation of
B800–850 complex may flow into the metabolism leading to the H2 production in PUC1.
䉷 2005 International Association for Hydrogen Energy. Published by Elsevier Ltd. All rights reserved.

Keywords: Photosynthetic spectral complexes; H2 production; Rhodobacter sphaeroides

1. Introduction inhibited if its component NifH is ADP-ribosylated by ADP-

Photoproduction of molecular hydrogen by Rhodospir-
illaceae is mediated by nitrogenase [1,2], which is reg-
ulated transcriptionally and post-transcriptionally by its
own end product, NH+ 4 [3–8]. Nitrogenase activity is
∗ Corresponding author. Tel.: +82 2 705 8459;
fax: +82 2 704 3601.
E-mail address: jgklee@sogang.ac.kr (J.K. Lee).
1 Present address: Department of Bacteriology, Division of Rick-
ettisial and Zoonotic Diseases, National Institute of Health, Seoul
122-701, Republic of Korea.
ribosyltransferase, DraT that is in turn activated by two PII-
like proteins, GlnB and GlnK, in response to ammonium
ion and darkness [4–6]. Expression of nifHDK, nitrogenase-
coding genes, is also negatively controlled in response to
ammonium ion; NifA, a transcriptional activator for nifHDK,
appears to be inhibited by GlnB and GlnK in the presence
of NH+ 4 , and is not expressed by the control of GlnB under
nitrogen-replete conditions [6–8].
The energy required for the nitrogenase-mediated H2
evolution is provided through photophosphorylation follow-
ing cyclic electron transfer [9,10]. Light is captured by two
light harvesting (LH) pigment–protein complexes, B800–850
and B875. The LH complexes trap light energy and transfer

0360-3199/$30.00 䉷 2005 International Association for Hydrogen Energy. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.ijhydene.2005.04.053
532 E.-J. Kim et al. / International Journal of Hydrogen Energy 31 (2006) 531 – 538
the excitation energy to a photochemical reaction center
(RC), the site of primary photochemistry. The pathway for
energy transduction is from B800–850 to B875 complex,
and to RC, in which B875 complex apparently serves as an
obligatory intermediate [11].
Previously, we isolated a purple photosynthetic bacterium
from mud samples collected at the seashore of Korea, which
showed higher production of H2 in comparison with the
laboratory strain, Rhodobacter sphaeroides 2.4.1 [12]. The
photoproduction of H2 by the isolate, which was identified
and deposited as R. sphaeroides KCTC 12085, was further
increased in proportion to light intensity following the sup-
pression of H2 uptake by mutation in hupSL coding for H2 -
uptake hydrogenase whose activity was regulated according
to the light intensity [12]. A fixed level of H2 production
was also elevated irrespective of the light intensities when
poly-
-hydroxybutyrate (PHB) formation was blocked by
mutation in phbC, reflecting the constitutive expression of
cellular PHB content under the conditions [12].
There have been reports of the improvement of H2 pro-
duction following changes in the level of spectral complexes
of R. sphaeroides [13,14]. In case of using solar energy, the
strong light (ca.
1.0 kW/m2 ) needs to be dispersed to el-
evate H2 production by R. sphaeroides [15]. However, self-
shading of cells due to their LH complexes could result in
reduction of light penetration [13,16] and therefore lower
the H2 production. In this work, we altered the level of LH
complexes of R. sphaeroides and examined the H2 produc-
tion with respect to light levels. The results in this work
suggest that the expenditure of cellular energy for the forma-
tion of B800–850 complex, rather than the complex-caused
shade on nearby cells in light path, appears to be a primary
reason to result in reduction of H2 production in wild type,
and B875 complex is obligatory for the efficient harvesting
of light energy even at the light level saturating cell growth.
2. Materials and methods
2.1. Bacterial strains and growth conditions
R. sphaeroides KCTC 12085 (KCTC: the Korean Collec-
tion for Type Cultures) [12] and R. sphaeroides 2.4.1 were
used as wild type strains. R. sphaeroides was grown aero-
bically or photoheterotrophically at 28 ◦ C in Sistrom’s suc-
cinate minimal medium [17] as previously described [18].
Light intensity for photoheterotrophic growth was measured
at the surface of culture vessels with a photometer (Li-Cor,
Inc., USA) [18], and was adjusted to 100 or 10 W/m2 . Mod-
ified Sistrom’s medium containing DL-malate (30 mM) and
L-glutamate (7 mM) was used for H2 accumulation [12].
Cell growth was monitored with a Klett–Summerson col-
orimeter (Manostat, USA) equipped with a KS-66 filter.
Escherichia coli was grown at 37 ◦ C in Luria–Bertani (LB)
medium. Kanamycin (Km), streptomycin (Sm), and specti-
nomycin (Sp) were used at 25, 50, and 50
g/ml for both
E. coli and R. sphaeroides. Ampicillin (Ap) and tetracycline
(Tc) were used at 50 and 20
g/ml, respectively, for E. coli,
whereas Tc was used at 1
g/ml for R. sphaeroides.
2.2. Construction of plasmids harboring puhA, pufBA, and
pucBAC of R. sphaeroides 2.4.1, and their maintenance in
R. sphaeroides KCTC 12085
The nucleotide sequence of R. sphaeroides 2.4.1 DNA
was obtained from a genome site at http://mmg.uth.tmc.edu/
sphaeroides/. A 903-bp HinfI-BamHI fragment (Fig. 1) ex-
tending from 40-bp upstream from puhA (reaction center
H polypeptide) [19] to its 80-bp downstream DNA was
filled-in and cloned into SmaI site of pBS. The recombi-
nant HindIII-EcoRI fragment from the plasmid was cloned
into pRK415 [20] to generate pRKH100, in which puhA
was situated in the same direction as tet promoter. The
DNA containing pufBA (B875-
and B875- polypeptides,
respectively) [21] (Fig. 2) was provided with pRKF100, a
recombinant pRK415 containing 1997-bp EcoRI-KpnI frag-
ment extending from the 216th residue of bchZ to the 117th
residue of pufL. The insert DNA of plasmid pRKF100 also
contains orfQ and pufK [22]. A 3386-bp XhoI-SacI DNA
(Fig. 3) containing pucBAC (B800–850-
, B800–850-
polypeptides, and an assembly protein for the B800–850
complex, respectively) [23] as well as its 465-bp upstream
and 980-bp downstream DNA was cloned into the SalI and
SacI sites of pRK415 to yield pRKC100.
Plasmids were mobilized from E. coli S17-1 into R.
sphaeroides through conjugation as described previously
[24]. Mating mixtures were plated aerobically on Sistrom’s
minimal medium containing appropriate antibiotics to ob-
tain isolated transconjugants.
2.3. Spectrophotometric assay of spectral complexes and
protein determination
Cell-free lysate of R. sphaeroides during exponential
growth was prepared as described previously [25] and
was used to examine the absorption spectra of photosyn-
thetic complexes with UV 2041-PC spectrophotometer
(Shimadzu, Japan). Amount of the B800–850 and B875
complexes were measured as described previously [11].
The amount of B800–850 complex was determined from
the spectral data by using A849.900 with an extinction
coefficient () of 96 mM−1 cm−1 , normalized for three
molecules of bacteriochlorophyll a per complex, whereas
the amount of B875 complex was calculated from A878.820
with  of 73 mM−1 cm−1 , normalized for two molecules
of bacteriochlorophyll a per complex. Protein content was
determined by the modified Lowry method [26]. The quan-
tification of LH complexes was repeated three separate
times, and the data shown in this work were reproduced
within the standard deviations less than 10%.
 E.-J. Kim et al. / International Journal of Hydrogen Energy 31 (2006) 531 – 538 533

BamH I

BamH I
Hinf I

Bgl II
Xho I

Xho I
Xho I
Pst I
100 bp

orf479 puhA orf213

Fig. 1. Restriction map of the R. sphaeroides 2.4.1 DNA containing puhA coding for RC-H polypeptide.
Hinc II
EcoR I

Sma I
Kpn I

Kpn I

Xho I
Acc I

Acc I

Acc I

Sma I
Pst I

Sal I
Mutant PUF1 ∆
200 bp
puf

bchZ orfQ K B A L M X

(A)

∆A = 0.2

WT

PUF1

350 Wavelength (nm) 900

(B)

Fig. 2. (A) Restriction map of the R. sphaeroides 2.4.1 DNA containing pufBA (B875-
and B875- polypeptides, respectively) is shown
and pufBA is in-frame deleted in mutant PUF1. Genes bchZ (BchZ subunit of chlorophyllide a reductase), orfQ (spectral complex assembly
protein [22]), pufLM (RC-L and RC-M polypeptides, respectively), and pufX (intrinsic membrane protein) are shown. (B) Absorption spectral
profiles are provided with cell-free extracts from R. sphaeroides KCTC 12085 (solid line; WT) and its mutant PUF1 (dotted line) grown
photoheterotrophically at 10 W/m2 .

2.4. Construction of pufBA-deleted and pucBA-deleted
mutants of R. sphaeroides KCTC 12085
Genes of pufBA and pucBA of R. sphaeroides KCTC
12085 were disrupted through homologous recombination
using appropriate DNA of R. sphaeroides 2.4.1. The pufBA-
deleted mutant PUF1 lacking of B875 complex was con-
structed as follows. The 220-bp DNA from the first nu-
cleotide of the 15th residues of pufB to the first nucleotide of
the 33rd residue of pufA was in-frame deleted from the 0.9-
kb AccI-KpnI DNA (Fig. 2A, thick line) of R. sphaeroides
2.4.1. A 404-bp DNA fragment immediately upstream from
the deletion region was PCR amplified with 5 primer F1
(5 -GTC AAC ACG CCG GTC CAT-3 ) and 3 primer R1
(5 -CTG CGC CTG CTG CAG TCC-3 , mutated sequence
underlined, unless otherwise noted) having the 14th and 15th
codons of pufB changed into PstI site (shown in bold). The
424-bp DNA fragment just next to the downstream from
the deletion region was also amplified using 5 primer F2
(5 -CCA CCA TGC ATC CTG CTG A-3́) having the 33rd
and 34th codons of pufA changed into NsiI site (shown in
bold), and 3 primer R2 (5 -CAA GGG CCG GCG GGT
AGA C-3 ). The upstream DNA fragment was digested by
AccI and PstI, and the downstream fragment was digested
534 E.-J. Kim et al. / International Journal of Hydrogen Energy 31 (2006) 531 – 538
BamH I
Xma III
Pvu II
Xmn I
Xho I
Pst I
Stu I
Mutant PUC1 ∆
puc
orf124 B A
(A)
350 Wavelength (nm)
(B)
Fig. 3. (A) Restriction map of the R. sphaeroides 2.4.1 DNA containing pucBA (B800–850-
and B800–850- polypeptides, respectively)
is shown and pucBA is in-frame deleted in mutant PUC1. pucC (assembly protein for B800–850 complex [23]) and nnr (LuxR-type
transcriptional regulator) are shown. (B) Absorption spectral profiles are provided with cell-free extracts from R. sphaeroides KCTC 12085
(solid line; WT) and its mutant PUC1 (dotted line) grown photoheterotrophically at 10 W/m2 .
by NsiI and KpnI. The 329-bp AccI-PstI upstream fragment
and 375-bp NsiI-KpnI downstream fragment were cloned
into the AccI and KpnI sites of pBS to generate pBSFD. Se-
quence analysis confirmed an in-frame deletion of pufBA as
well as no other mutations in the amplified DNA. The insert
DNA of pBSFD was then subcloned into SacI and SphI sites
of a suicide plasmid pLO1 [27,28]. The resulting plasmid
pLO1-pufBA, which harbored aph (Kmr ) and sacB, was
mobilized from E. coli S17-1 into R. sphaeroides KCTC
12085 through conjugation as described [24]. The Kmr ex-
conjugant, which was generated from single crossover, was
isolated and subjected to segregation to double-crossover
recombination showing Kms phenotype on sucrose (15%)-
containing Sistrom’s minimal medium. As a result, five of
such recombinants devoid of B875 complex were obtained.
The chromosomal structures of the mutants were examined
for the correct deletion by genomic Southern hybridization
analysis [29], and one recombinant PUF1 was analyzed fur-
ther.
In order to construct the pucBA-deleted mutant PUC1,
the 267-bp XmnI-XmaIII DNA spanning from pucB to pucA
was removed from 1.34-kb PstI-PvuII fragment (Fig. 3A,
thick line) of R. sphaeroides 2.4.1. The resulting recom-
Xho I
Xho I
Xho I
Sph I
Sac I
Pst I
Pst I
Pst I
200 bp
C nnr
∆A=0.2
WT
PUC1
900
binant DNA fragment was cloned into pLO1 to generate
pLO1-pucBA, which was subsequently mobilized from E.
coli S17-1 into R. sphaeroides KCTC 12085 through con-
jugation as described previously [24]. Double-crossover re-
combinations were selected as described above for PUF1.
As a result, four of such recombinants devoid of B800–850
complex were obtained. The genomic DNA of the recombi-
nants was examined for correct deletion of pucBA by South-
ern hybridization analysis [29], and one recombinant PUC1
was analyzed further.
2.5. Measurement of hydrogen gas
R. sphaeroides was inoculated into a modified Sistrom’s
medium for H2 evolution in a homemade serum bottle
(60 ml) having a side arm (1.2 × 11; diameter × height in
centimeters), followed by gassing with argon as described
previously [12]. Cells were contained in the side arm of the
bottle, and the arm tube was stood in a tube rack in front of
light box. The gas phase (100–300
l) was withdrawn inter-
mittently during photoheterotrophic growth and analyzed
for H2 by GC-17A gas chromatograph (Shimadzu, Japan)
equipped with a thermal conductivity detector and a col-
 E.-J. Kim et al. / International Journal of Hydrogen Energy 31 (2006) 531 – 538 535

umn containing 60/80 mesh Molecular Sieve 5A (Supelco, and 100 W/m2 , and compared with those of the cells con-
USA). Argon was used as a carrier gas. The analyses of taining pRK415 as controls. As observed previously [12],
H2 accumulation in this work were performed at least three H2 production reached the maximum level at the stationary
separate times, and the data presented here were reproduced phase of cell growth (Fig. 4, A1 and A2). All the cells grew
within standard deviations of 10–15%. with doubling time between 5 and 6 h (Fig. 4, B1 and B2).
Therefore, the light level as much as 10 W/m2 is saturating
cell growth.
3. Results and discussion The cells containing the recombinant plasmids produced
less H2 compared with the cells containing pRK415 (Fig.
3.1. Photosynthetic spectral complexes are increased by 4, A1 and A2). Although it remains to be determined how
maintaining the plasmids harboring DNA encoding puhA, the H2 production is lowered in the order of cells con-
pufBA, or pucBAC in trans in R. sphaeroides taining plasmids pRKH100, pRKF100, and pRKC100, the
presence of additional spectral complexes appears to result
Plasmids pRKH100, pRKF100, and pRKC100 contain in reduction of the maximum level of H2 production by
the insert DNA encoding puhA (Fig. 1), pufBA (Fig. 2), approximately 10–50% in comparison with those of the
and pucBAC (Fig. 3), respectively, which are derived from control cells at the light intensities examined (Fig. 4, A1
R. sphaeroides 2.4.1. Each plasmid was mobilized into R. and A2). The same decrease was also observed with the H2
sphaeroides KCTC 12085, and the transconjugants were accumulation rates (data not shown), which were in parallel
grown photoheterotrophically at 10 and 100 W/m2 . The cel- with the maximum levels of H2 production as described
lular level of B875 and B800–850 complexes were measured previously [12]. The results suggest that the low level of
and compared with those of cells containing the vector plas- H2 production by maintaining the recombinant plasmids in
mid pRK415 (Table 1). The B875 complex was increased cells cannot be ascribed to light-limiting due to the shadow
by approximately 30–50% only when pRKF100 containing by higher level of spectral complexes, because neither the
pufBA was provided in trans. No such increase was observed H2 production nor cell growth were enhanced at the brighter
by either pRKH100 or pRKC100. However, all the transcon- light 100 W/m2 (Fig. 4, A2 and B2) that should have di-
jugants containing the recombinant plasmids showed an in- minished the shade if any. The energy-wasting metabolism
crease of the level of B800–850 complex by approximately leading to the formation of surplus spectral complexes
20–40%. rather appears to be a primary reason for the decreased H2
production.
3.2. Increase in cellular level of photosynthetic spectral
complexes decreased the photoheterotrophic production 3.3. Removal of B875 complex not only slowed cell
of H2 growth but also decreased H2 production even at light
(
10 W/m2 ) saturating the growth of wild type
R. sphaeroides KCTC 12085 containing pRKH100,
pRKF100, and pRKC100 were examined for the photo- Since an increase in cellular level of the photosynthetic
heterotrophic production of H2 at light intensities of 10 spectral complexes decreased the photoheterotrophic pro-
duction of H2 (Fig. 4), it was examined whether removal of
either B875 or B800–850 complex results in elevation of H2
Table 1 production. The DNA encoding pufBA was in-frame deleted
Relative level of LH complexes of the wild-type cells carry-
from the chromosome of R. sphaeroides KCTC 12085. The
ing pRKH100 (puhA), pRKF100 (pufBA), pRKC100 (pucBAC),
resulting mutant PUF1 (Fig. 2A) did not show any B875
and pRK415 (vector plasmid control), which were grown photo-
heterotrophically at 10 and 100 W/m2 complex under the photoheterotrophic conditions (Fig. 2B).
The level of B800–850 complex of PUF1 was not different
Light Strains Level of LH complex from the wild type although the carotenoid level appears to
intensity carrying (nmole/mg of protein) be lowered (Fig. 2B).
(W/m2) plasmids The H2 production and cell growth of wild type at
B875 B800–850 10 W/m2 were not much different from those observed at
10 KCTC12085 (pRK415) 11.9 ± 0.1 13.6 ± 0.6 100 W/m2 (Fig. 5, A and B). The growth of PUF1 under
KCTC12085 (pRKH100) 12.3 ± 0.6 16.4 ± 1.0 aerobic conditions is the same as that of wild type (data not
KCTC12085 (pRKF100) 17.8 ± 0.1 17.3 ± 0.8 shown). However, PUF1 grew with a doubling time of ap-
KCTC12085 (pRKC100) 10.7 ± 0.5 17.8 ± 0.1 proximately 13 h at 10 W/m2 and its maximum cell density
(KU) went up to approximately 80% of that of wild type
100 KCTC12085 (pRK415) 11.8 ± 0.5 9.1 ± 0.6 which doubled in every 5 h under the same conditions (Fig.
KCTC12085 (pRKH100) 11.7 ± 0.9 12.9 ± 0.1
5, B1). PUF1, though with 7-h doubling time, grew up to
KCTC12085 (pRKF100) 15.1 ± 0.6 11.4 ± 0.4
the same KU as that of wild type at 100 W/m2 (Fig. 5, B2).
KCTC12085 (pRKC100) 9.3 ± 0.2 12.2 ± 0.3
The results suggest that B875 complex is essential for the
536 E.-J. Kim et al. / International Journal of Hydrogen Energy 31 (2006) 531 – 538

2000 10 W/m2 2000 100 W/m2

H2 (µl/ml culture)

H2 (µl/ml culture)
1500 pRK415 1500 pRK415
pRKH100 pRKH100
1000 pRKF100 1000 pRKF100
pRKC100
pRKC100
500 500

0 0
0 40 80 120 0 40 80 120
(A1) Time (h) (A2) Time (h)

1000 1000
KU

KU
100 100

10 10
0 40 80 120 0 40 80 120
(B1) Time (h) (B2) Time (h)

Fig. 4. H2 accumulation (A1, A2) and cell growth (B1, B2) of R. sphaeroides KCTC 12085 containing pRK415 (square), pRKH100
(diamond), pRKF100 (circle), and pRKC100 (triangle) under the photoheterotrophic conditions at 10 W/m2 (A1, B1) and 100 W/m2 (A2, B2).

2500 10 W/m2 2500 100 W/m2

H2 (µl/ml culture)

H2 (µl/ml culture)

2000 PUC1 2000

1500 1500 PUC1

WT WT
1000 1000
PUF1
500 500
PUF1
0 0
0 50 100 150 0 50 100 150
(A1) Time (h) (A2) Time (h)

1000 1000
KU
KU

100 100

10 10
0 50 100 150 0 50 100 150
(B1) Time (h) (B2) Time (h)

Fig. 5. H2 accumulation (A1, A2) and cell growth (B1, B2) of R. sphaeroides KCTC 12085 (square; WT), and its two mutants, PUC1
(diamond) and PUF1 (triangle), under the photoheterotrophic conditions at 10 W/m2 (A1, B1) and 100 W/m2 (A2, B2).

efficient light harvesting and transfer of excitation energy
to RC. The growth of PUF1 was fully complemented with
pRKF100 (data not shown), which suggests no additional
mutation(s) in the mutant except for pufBA deletion.
The H2 production of PUF1 during the photo-
heterotrophic growth at 10 W/m2 amounted to only 30% of
the wild-type level (Fig. 5, A1). PUF1, however, produced
H2 up to the 80% level of wild type at 100 W/m2 (Fig. 5,
A2), which is consistent with its growth restoration at the
brighter light (Fig. 5, B). Thus, the B875 complex needs to
be present for efficient cell growth and H2 production of R.
sphaeroides.
 E.-J. Kim et al. / International Journal of Hydrogen Energy 31 (2006) 531 – 538
3.4. Removal of B800–850 complex increased the H2
production at the light level saturating the growth of wild
type
The B800–850 complex was removed by deleting pucBA
on the chromosome of R. sphaeroides KCTC 12085 to gen-
erate mutant PUC1 (Fig. 3A) which grew as wild type under
aerobic conditions (data not shown). No B800–850 complex
was found in PUC1 grown photoheterotrophically (Fig. 3B).
The level of B875 complex of PUC1 was not different from
wild type. PUC1 grew with a doubling time of 6 h during
photoheterotrophic growth at 10 W/m2 , and the maximum
KU was approximately 20% lower than that of the wild type
as observed with PUF1 (Fig. 5, B1). However, PUC1 grew
like wild type at 100 W/m2 (Fig. 5, B2), which indicate
that the B800–850 complex is needed for the efficient light
harvesting at light dimmer than 10 W/m2 , and the complex
requirement for the photoheterotrophic growth can be com-
pletely abolished if cells are exposed to the brighter light.
The H2 production of PUC1 during photoheterotrophic
growth at 10 W/m2 was increased up to nearly twofold in
comparison with that of wild type (Fig. 5, A1). The 30–40%
increase of H2 production of the mutant was also observed in
the same comparison at 100 W/m2 (Fig. 5, A2) although it re-
mains to be determined how its H2 production at 100 W/m2
is lower than that observed at 10 W/m2 . Thus, the H2 pro-
duction of PUC1 was not increased in proportion to the
light intensity (Fig. 5, A1 and A2). The same was true for
wild type. The results suggest that the cellular energy for
H2 production of wild type and PUC1 were not limited at
10 W/m2 , but the energy that may be saved by the lack of
B800–850 complex of PUC1 rather appears to flow into the
metabolism leading to the enhanced production of H2 under
the photoheterotrophic conditions.
4. Concluding remarks
1. Increase in the cellular level of B800–850 and B875 com-
plexes of R. sphaeroides resulted in decrease of H2 pro-
duction.
2. The B875 complex needs to be present in cell mem-
brane for efficient cell growth and H2 production of R.
sphaeroides.
3. Removal of the most peripheral B800–850 complex in-
creased H2 production by up to approximately twofold
in comparison with its parental cell at the light level sat-
urating the growth of wild type.
Acknowledgements
This Research was supported by the Hydrogen En-
ergy R&D Center, one of the 21st Century Frontier R&D
537
Programs, funded by the Ministry of Science and Technol-
ogy of Korea.
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