JOURNAL OF BIOSCIENCE AND BIOENGINEERING © 2005, The Society for Biotechnology, Japan

Vol. 100, No. 5, 481–488. 2005

DOI: 10.1263/jbb.100.481

REVIEW
Applications of Photosynthetic Bacteria for Medical Fields
Ken Sasaki,1* Masanori Watanabe,1 Yoshito Suda,1 Akihiro Ishizuka,2
and Napavarn Noparatnaraporn3
Materials Science and Engineering, Graduate School of Engineering, Hiroshima Kokusai Gakuin University,
6-20-1 Nakano, Akiku, Hiroshima 739-0321, Japan,1 Research and Development Center, Cosmo Oil Co., Ltd.,
1134-2 Gongendo, Satte-shi, Saitama 340-0193, Japan,2 and Department of Microbiology,
Faculty of Science, Kasetsart University, Chatuchak, Bangkok 10900, Thailand 3

Received 6 May 2005/Accepted 19 July 2005

The medical applications of photosynthetic bacteria are summarized. Photosynthetic bacteria

can produce various types of physiological active substance such as vitamin B12, ubiquinone (co-
enzyme Q10), 5-aminolevulinic acid (ALA), porphyrins and RNA. In particular, photosynthetic
bacterial ALA was commercially applied to cancer diagnosis and treatment. Recently, ALA has
been applied to the treatment of acne vulgaris and the suppression of the inflammatory response
to coronary and iliac injuries. In addition, the recent applications of RNA from a marine photo-
synthetic bacterium as a medical supplement for immune improvement and suppression of infec-
tion are described. Furthermore, the feasible application of a biopolymer consisting of RNA from
a photosynthetic bacterium as a drug delivery system (DDS) to cancer treatment is described.

[Key words: photosynthetic bacteria, 5-aminolevulinic acid, porphyrin, RNA, drug delivery]

Photosynthetic bacteria, which are common microorga- medical fields.

nisms in the natural environment, have been applied in the
field of environmental protection such as in the treatment of
sewage, and domestic and restaurant wastewaters (1, 2), and
the bioremediation of sediment mud polluted with organic
matters (3). Some photosynthetic bacteria such as Rhodo-
spirillacea can grow with a relatively high growth rate un-
der an aerobic dark condition without utilization of light en-
ergy. They can utilize various types of organic matter as car-
bon and energy substrates; therefore, they are applied to en-
vironmental protection (1). On the other hand, photosyn-
thetic bacteria produce relatively large amounts of physio-
logical active substances such as vitamin B12, ubiquinone
(coenzyme Q10), 5-aminolevulinic acid (ALA), porphyrins
and RNA (1). In particular, coenzyme Q10 and ALA have
been prepared and commercialized on the basis of produc-
tion of such components by photosynthetic bacteria (1, 4,
5).
Application studies of photosynthetic bacteria for medi-
cal fields were reviewed previously (5). In this paper, we re-
view the recent applications of photosynthetic bacteria in
* Corresponding author. e-mail: sasaki@hkg.ac.jp
phone: +81-(0)82-820-2570 fax: +81-(0)82-820-2560
Abbreviations: ALA, 5-aminolevulinic acid; D, dilution rate; DCW,
dry cell weight; DDS, drug delivery system; DO, dissolved oxygen;
EPS, extracellular polymeric substance; 5FU, 5-fluoro-1-urasil; HPLC,
high-pressure liquid chromatography; MLSS, mixed liquor suspended
solid; ORP, oxidation reduction potential; PDT, photodynamic ther-
apy; PPIX, protoporphyrin IX; Tegaful, 5-fluoro-1-(tetrahydro-2-fur-
furyl) urasil.
PRODUCTION OF VITAMIN B12
Vitamin B12 has been used in treating anemia and neuritis
and as an eye lotion. Recently, its applications as health
food supplements have become popular. Another applica-
tion of vitamin B12 is as a growth-promoting factor in feed-
stock for animals (1, 5). The commercial production of
vitamin B12 was biologically carried out using Propionibac-
terium; however, photosynthetic bacteria are another potent
producer of vitamin B12. A photosynthetic bacterium, Rho-
dobacter sphaeroides P47, produced 75 µg/g dry cells under
aerobic dark culture condition. Other photosynthetic bac-
teria such as R. capsulatus and R. gelatinosa produced 21–
33 µg/g dry cells under aerobic dark culture condition (1).
Under anaerobic light culture condition, R. sphaeroides pro-
duced 87 µg/g dry cells of vitamin B12 which was almost
twice the production compared with that under aerobic dark
culture condition. However, aerobic dark cultivation is prac-
tical for mass cultivation. Vitamin B12 can be synthesized
through a pathway common to porphyrin, chlorophyll and
haem via 5-aminolevulinic acid (ALA). The Shemin-path-
way (C-4 pathway) is the main pathway operating in pho-
tosynthetic bacteria for vitamin B12 biosynthesis in which
glycine and succinate are the precursors of vitamin B12 bio-
synthesis (1, 6) in contrast with the C-5 pathway in Propi-
onibacterium in which glutamate is the precursor. Although
the vitamin B12 content of photosynthetic bacteria is not so
high compared with that of Propionibacterium, 1.58 mg/l

481
482 SASAKI ET AL.
(75 µg/g dry cells) vitamin B12 was produced from R.
sphaeroides P47 culture in pineapple juice waste. This ob-
servation is of practical interest in producing vitamin B12
from low-cost food waste considering the use of recycled
renewable resources (1). Cells containing vitamin B12 can
be directly utilized as feedstock or food supplements since
their carbon source is pineapple which is a safe food. Such
health food applications will be developed for the medical
application of vitamin B12 using photosynthetic bacterial
cells.
COENZYME Q10 PRODUCTION
Coenzyme Q10 has been used in treating heart diseases
for more than 30 years (7). Coenzyme Q10 has variety of
physiological activities indicated for hypertension, brain vas-
cular injury, anemia, muscle dystrophy and alveolar pyor-
rhea. Recently, coenzyme Q10 has been used not only as a
medicine but also as food supplements because of its vari-
ous physiological activities.
The commercial production of coenzyme Q10 started
about 25 years ago mainly by using photosynthetic bacteria
and the efficiency in such production has improved recently.
Because photosynthetic bacterial cells contain large amounts
of coenzyme Q10. In particullar, anaerobic light culture of
Rhodospirillacae, coenzyme Q10 production was twofold
higher than that in aerobic dark culture (1). However, aero-
bic coenzyme Q10 production has been applied to the mass
culture of cells without light irradiation which is costly for
practical production. For example, R. sphaeroides produced
1.5 mg/l (0.7 mg/g dry cells) coenzyme Q10 and R. gelati-
nosus produced 1.6 mg/l (0.8 mg/g dry cells) of coenzyme
Q9 under aerobic dark cultivation (1). R. gelatinosus cells
produced mainly coenzyme Q9 (not Q10). Under anaerobic
light cultivation, R. sphaeroides P47 produced 3.0 mg/g dry
cells (1.7 mg/l Q10). We found that coenzyme Q10 produc-
tion was enhanced under microaerobic dark cultivation,
which was the aerobic culture with an almost nil level of
dissolved oxygen (DO) in the culture broth (1). A relatively
high cell mass and a high coenzyme Q10 level could be pro-
duced simultaneously in a microaerobic culture. Coenzyme
Q10 production was also affected by controlling oxidation
reduction potential (ORP). When ORP was controlled at a
low level, coenzyme Q10 production was enhanced. At a
−200 mV ORP, coenzyme Q10 production reached 13.5 mg/g
dry cells (8). Respiration quotient also affected coenzyme
Q10 production (8).
The utilization of Coenzyme Q10 as a food supplement is
now being developed. Recent medical functions of coen-
zyme Q10 such as insulin-like functions and suppressive
effect in diabetes neurosis have been reported (7). The com-
mercial production of coenzyme Q10 is now being devel-
oped in Japan and China.
ALA PRODUCTION
ALA is a well-known intermediate of tetrapyrrole biosyn-
thesis such as porphyrin, haem and vitamin B12 (6). How-
ever, few research studies about ALA production and its
physiological characterization had been reported until 1980
J. BIOSCI. BIOENG.,
because of difficulties in ALA production as a commercial
reagent. The chemical synthesis of ALA has been rather dif-
ficult until now due to the many steps involved in it (9).
As for the biological production of ALA, some reports
were previously presented in the 1980s. ALA production
(also called formation) was carried out by adding levulinic
acid as an inhibitor of ALA dehydratase and extracellular
ALA accumulation. However, the extracellular accumula-
tion level of ALA was quite low for practical applications
(0.1–1.0 µM). We observed large amounts of ALA excre-
tion (about 4.2 mM) by the photosynthetic bacterium, R.
sphaeroides with the intermittent addition of levulinic acid
during cultivation (10). By various improvements in the pro-
duction procedure, practical methods of ALA production by
photosynthetic bacteria have been established (11, 12).
ALA was first used in the field of agriculture as a biode-
gradable herbicide and growth- promoting factor for plants.
These applications were reviewed in our previous paper (6).
Recently, medical applications of ALA have been devel-
oped. Traditionally, for a long time, ALA was used for the
diagnosis of heavy- metal poisoning. However, Kennedy et
al. used ALA for cancer treatment by laser irradiation tech-
nology (13). This form of treatment was called photody-
namic therapy (PDT) (13–15). For example, ALA-contain-
ing cream was applied to cancer-affected areas of the skin.
ALA accumulated in the cancer cells and protoporphyrin IX
(PPIX) was produced. After 3–4 h, laser irradiation was
carried out. The cancer cells were killed by singlet oxygen
produced by PPIX. But the normal cells were not damaged
because PPIX was not formed in the cells (13). This mecha-
nism is the same as that of plant herbicides (16). With the
use of ALA, 86% of the skin cancer could be treated. Hae-
matoporphyrin derivatives are frequently used for PDT. How-
ever, haematoporphyrin derivatives take time to accumulate
in cancer cells. In addition, the side effects of a haematopor-
phyrin derivatives, such as light injuries are serious prob-
lems in the use of PDT. By using ALA, such side effects
will not be produced. PDT using ALA was applied to the
treatment of not only skin cancer but also oral, esophageal,
colon, duodenal, pancreatic and bladder cancers (6).
One of the recent developments of ALA application is in
the area of photodynamic diagnosis (14, 15). As shown in
Fig. 1, ALA was used in the diagnosis of glioma cells (brain
tumor cells) during surgery. The glioma cells were detected
by their red fluorescence with violet-blue light (395–415
nm) irradiation, thereby facilitating surgery. PPIX mainly
accumulated in the tumor cells, while normal brain cells did
not accumulate PPIX (nonfluorescent). Good diagnosis and
operative results were achieved by this procedure (15).
Another recent application of ALA in the medical field
was in the PDT of intractable acne vulgaris for laser irradia-
tion. Four days after irradiation, the lesion formed a very
thin crust and was completely healed in 10 d (17). Thus,
ALA applications to laser irradiation is very promising and
must be developed. Furthermore, ALA was applied to the
suppression coronary and iliac artery injuries in response to
PDT (18). ALA inhibits neointimal hyperplasia in injured
arteries in animals. Application still at the animal research
or experimental level, however, it may soon be applied to
humans. Further applications of ALA are now under inves-
VOL. 100, 2005 MEDICAL APPLICATIONS OF PHOTOSYNTHETIC BACTERIA 483

FIG. 1. Photodynamic diagnosis of glioma (brain tumor) during surgery. Accumulated PPIX (protoporphyrin IX) from ALA in tumor cells
fluoresces under violet-blue light irradiation, thereby simplifying the distinction between glioma cells and normal cells (cited from Kaneko et al.
[14]).

tigation in various medical fields.

PORPHYRIN PRODUCTION
Porphyrin are well-known intermediates of the biosyn-
thesis of tetrapyrroles such as chlorophyll, haem and vita-
min B12. Porphyrin is widely commercialized as a medicine
for liver diseases, cancer diagnosis and cancer treatment.
For example, haematoporphyrin is frequently used for PDT
of cancer. Up to now, porphyrin derivatives have been pro-
duced from discharged blood from cows and other animals.
However, mad cow disease (bovine spongiform encephalo-
pathy, BSE) and other animal diseases have made prophyrin
production from animal blood less acceptable for medical
applications. The microbial production of porphyrin has
therefore become more attractive.
As for the microbial production of porphyrin, photosyn-
thetic bacteria are the most potent microorganisms because
R. sphaeroides can produce 10–100 mg/l of porphyrin extra-
cellularly (19). Ishii et al. (20) reported the production of
115 mg/l porphyrin using immobilized photosynthetic bac-
teria with light illumination. Kojima et al. reported a 635
mg/l coproporphyrin and uroporphyrin production by a mu- FIG. 2. Aerobic porphyrin production in dark by R. sphaeroides
IFO 12203 and mutant R. sphaeroides CR386 in GGY2 medium. Culti-
tant strain of Arthrobacter hyalinus under aerobic dark cul- vation was carried out in a 2-l jar fermentor (TITEC, CTB-33) with 1 l
tivation (21). Aerobic dark cultivation is attractive from a of medium at 30°C. pH was controlled to 7.0 ±0.1. DO >7.0 mg/l. ALA-
practical point of view. HCl (2 g/l) was added at the late exponential phase. (a) Porphyrin for-
We have been studying practical methods of porphyrin mation; (b) cell growth; (c) ALA concentration in the medium. Closed
lozenges and closed squares indicate IFO12203 and CR386, respec-
production using photosynthetic bacteria under aerobic dark tively.
culture condition and we have isolated a mutant strain of R.
sphaeroides CR386 produced by UV irradiation. In Fig. 2, a
comparison of porphyrin production between two strains of duced uroporphyrin at 1.0%, coproporphyrin III at 93.9%
photosynthetic bacteria is shown under aerobic dark condi- and protoporphyrin III at 5.1% (22). In A. hyalinus CYS,
tion using GGY2 medium (glucose and yeast extract me- 30% uroporphyrin and 70% coproporphyrin were extracel-
dium). R. sphaeroides IFO12203 which is a type culture of lularly produced simultaneously (21). Therefore, relatively
R. sphaeroides produced almost no porphyrin extracellu- complex separation procedures are required to obtain pure
larly under aerobic dark cultivation; however, CR386 pro- coproporphyrin III which is a key intermediate in the syn-
duced about 15 mg/l of porphyrin under a completely aero- thesis of porphyrin derivatives. The use of CR389 is more
bic condition (DO >7 mg/l) (22). The advantage of porphy- advantageous considering that no practical separation pro-
rin production by CR386 was that almost 100% copropor- cedures that require a high cost of treatment.
phyrin III and less than 0.1% uroporphyrin III and protopor- In addition, we also found that porphyrin extracellular
phyrin III were produced. In contrast to this, IFO12203 pro- production is affected by the DO level of the culture condi-
484 SASAKI ET AL.
FIG. 3. Effects of strict control of DO (± 0.2 mg/l) for aerobic por-
phyrin production in dark by R. sphaeroides CR386 in GGY2 medium.
Cultivations were carried out in the same manner as that in the jar fer-
mentor shown in Fig. 2 with the DO controlled to 0, 1.0, 2.0, 3.0 mg/l.
(a) Porphyrin formation; (b) cell growth; (c) ALA concentration in
the medium; (d) DO in the medium. Closed lozenges, closed squares,
closed triangles and asterisks indicate DO controlled to 0, 1.0, 2.0 and
3.0 mg/l, respectively.
tion. As shown in Fig. 3. DO influenced porphyrin produc-
tion. At a controlled DO level of about 2.0 ±0.2 mg/l, por-
phyrin (coproporphyrin III) production reached 56.3 mg/l.
At high and low DO controlled levels, porphyrin production
was not so high (Fig. 3). The value of 56.3 mg/l is relatively
low compared with that of A. hyalinus CYS, but the culture
time is shorter and a separation procedure is unnecessary for
obtaining coproporphyrin III. Therefore, this production is
advantageous for the production of labeled porphyrin which
can be used for medical research. In addition, by immobiliz-
ing the cell system, higher porphyrin production may be
possible under aerobic dark culture condition. These tech-
nologies (mutant utilization and DO control) may be applied
to the practical production of various types of porphyrin.
RNA PRODUCTION
RNA is an attractive source of 5′-ribonucleotides for use
as a flavor enhancer in the food industry (23). Recently,
RNA production has attracted another interest as a dietary
source of pyrimidine and purines for human immune func-
tions (24). In Table 1, the effects of ribonucleotides and
RNA on the immune response of humans have been re-
ported (24). For example, the suppression of the rejection
J. BIOSCI. BIOENG.,
TABLE 1. Effects of ribonucleotides in medical applications
Intestinal administration
Specialization of organ; growth of organ
Protection from infection; protection from complication
(shortening of hospitalization period)
Improvement of immune system (protection from infection;
suppression of rejection after transplantation)
Intravenous administration
Improvement of protein metabolism
Enhancement of reclamation of liver cells
Protection of mucous membrane of intestine
Improvement of immune function (protection from infection;
suppression of rejection after transplantation)
Improvement of energy metabolism
Reduction of obstacle of blood flow
Biochemical modulation
In vitro effects
(in vitro applications for basic medical research experiments)
Cultivation of liver cells
Cultivation of tumor cells
Growth of macrophage
Modified from Usami and Saitoh (24).
reaction after transplantation and suppression of Staphylo-
coccus aureus (MRSA) infection are quite attractive effects
because in particular some antibiotics are not effective in
suppressing MRSA infection. In addition to the effects on
the immune response of humans, other medical applications
of RNA summarized in Table 1 (24). For example, improve-
ment in protein and energy metabolism, enhancement of the
reclamation of liver cells, protection and maintenance of
mucous membrane of the intestine and reduction in the ob-
stacle of blood flow are reported.
Ribonucleotides are usually produced by the enzymatic
hydrolysis of RNA from waste yeast cells such as brewed
beer yeast (25). Vidotto et al. reported the results of an ef-
fective RNA production by Candida albicans using dif-
ferent concentrations of ammonium in the medium (26). We
have found recently that the photosynthetic bacterium,
Rhodovulum sp. PS88 strain can produce extracellular poly-
meric substances (EPSs) in the surface of its cells (27).
These EPSs contain large amount of RNA (about 60 mg/g
cells), however their DNA content was low. EPS also con-
tains about 45 mg protein/g cells (27). The PS88 strain has a
self-flocculating activity and we conclude that the RNA was
the major element for flocculation due to the deflocculation
by RNase treatment in contrast with the lack of defloccula-
tion by protease treatment (27).
The mass production of RNA by the PS88 strain was
examined using an aerobic continuous culture system. At a
high dilution rate (D =growth rate) in continuous culture,
extensive RNA synthesis can be expected (28). The PS88
strain allowed for easier cell separation by self-flocculation.
As shown in Fig. 4, aerobic dark continuous cultivation in a
1-l jar fermentor (500 ml medium) with 1 vvm and 300 rpm
(DO > 3 mg/l) culture conditions was carried out. RNA pro-
duction at a steady state condition was measured. The MLSS
corresponding to dry cell weight (DCW) plus EPS was al-
most constant at various dilution rates. The maximum RNA
production was observed at a dilution rate (D) of 0.32 h–1
which was 460 mg/l of RNA. At a high dilution rate of more
than 0.32 h–1, RNA concentration decreased and productiv-
VOL. 100, 2005
FIG. 4. Steady-state results of growth and RNA production by
Rhodovulum sp. PS88 in continuous culture. One-liter jar fermentor
(500 ml of medium) was used for the production of RNA. Acetate me-
dium was used and DO was controlled to more than 3.0 mg/l. Temper-
ature and pH were kept at 20 ± 0.5°C and 8.0± 0.1, respectively. The
dilution rates (D) ranged from 0.20 to 0.55 h–1. Mmixed liquor sus-
pended solid (MLSS) indicate dry cell weight (DCW) plus extracellu-
lar polymeric substances (EPS). (a) Productivity of RNA; (b) RNA in
the culture medium; (c) MLSS and EPS in the culture medium. Closed
circles, open circles, closed triangles, closed squares, open triangles
indicate productivity of RNA, RNA in MLSS (cells and EPS), RNA in
the broth, MLSS and EPS in the broth, respectively.
ity was almost constant (150–170 mg RNA/l/h). In addition,
RNA was produced not only in the EPS but also in the cells.
For example, as shown in Table 2, intracellular RNA was
about threefold higher than that of EPS in every D. The max-
imum RNA content in the cells was 134 mg/g cells (MLSS).
With EPS, the maximum RNA content was 203 mg/g MLSS
(134 mg/MLSS plus 69 mg/EPS) at D of 0.32 h–1. This value
was two- to threefold greater than that of yeast RNA pro-
duction (26). Such high RNA production was not reported
so far. These observations suggest that the mass production
of RNA by photosynthetic bacteria that have the self-floc-
culation activity is possible. Some photosynthetic bacteria
of Rhodospirillacea have the self-flocculation activity.
MEDICAL APPLICATIONS OF PHOTOSYNTHETIC BACTERIA 485
TABLE 2. Distributions of RNA in cells , EPS and culture broth at
each steady state of continuous culture of Rhodovulum sp. PS88
RNA in MLSS RNA
D (1/h) Intracellular EPS in broth
mg/l mg/MLSS mg/l mg/MLSS mg/l
0.10 171 102 121 49 96.7
0.22 284 129 129 46 82.9
0.32 266 134 193 69 66.5
0.44 192 113 167 65 49.5
0.58 191 124 113 53 44.9
MEDICAL APPLICATION OF EPS
The photosynthetic bacterium, Rhodovulum sp. PS88 pro-
duces EPS which contains large amounts of RNA on the
surface of the cells. EPS also contains proteins and is a bio-
polymer which has sticky characteristics. In connection
with this, some biopolymers have been applied as a drug de-
livery system (DDS) like chitosan gel. Chitosan gel has
been applied to cancer chemotherapy (29), preventing re-
stenosis and gene therapy as a new DDS (30, 31). Biopoly-
mers such as chitosan gel and EPS from PS88 have advan-
tages over chemically synthesized polymers because these
are safe to use, biodegradable and have low side effects
being natural biopolymers (29, 30). In addition, RNA has
immune suppressive effects and exerts protection against
MRSA infection as shown in Table 1. Therefore, the feasi-
bility of the application of EPS from PS88 strain to DDS
was investigated.
First, the possibility of drug adsorption under various pHs
was examined using an anticancer agent model, 5-fluoro-
1-uracil (5FU), which is a popular agent for testing DDS ac-
tivity. EPS was prepared as described previously (27). EPS
reaction mixture (50 ml) in 5 mM phosphate buffer which
contained 10 mM magnesium ions and 10 mg of 5FU were
mixed and incubated for 20–120 min at 20°C under recipro-
cal shaking (100 rpm, 10 cm amplitude). Residual 5FU was
measured by HPLC with a Shim-pack STR ODSII column.
As shown in Fig. 5, the maximum amount of 5FU de-
crease was observed during the 40-min incubation at each
pH. At pH 8.0, the maximum amount of drug decrease was
observed (Fig. 5a). This is a favorable characteristics be-
cause pH 8.0 is close to that of human blood. On the other
hand, in the 5FU and Mg mixture also examined as control
experiment (Fig. 5b), there was an approximately 50% de-
crease in 5FU level by conjugation with Mg. Consequently,
the adsorbed 5FU to EPS (EPS–5FU conjugate) was esti-
mated as shown in Fig. 5c. The maximum adsorption was
6.8 mg 5FU/l EPS reaction mixture at pH 8.0. This level is
similar to that of chitosan gel (29, 30).
Another anticancer reagent model 5-fluoro-1-(tetrahydro-
2-furfuryl) uracil (Tegaful) was also examined in the same
manner by HPLC. As shown in Fig. 6, the adsorption trend
was the same as that of 5FU. Approximately 14% (27.9 mg
Tegaful/l EPS reaction mixture) was adsorbed as EPS-
Tegaful conjugate. Thus, the EPS–gel–drug conjugate of 5FU
and Tegaful consist of 6.8 mg of 5FU and 27.9 mg of
Tegaful per EPS . These values were quite similar to that of
chitosan gel, which is an excellent biopolymer for DDS (29,
486 SASAKI ET AL.
FIG. 5. Adsorption of 5FU onto EPS gel under different pHs. (a)
EPS, Mg2+ and 5FU contained in reaction mixture. (b) Mg2+ and 5FU
contained in reaction mixture as control experiment. (c) Estimated ad-
sorbed 5FU on EPS gel. The initial 5FU concentration and temperature
in the reaction mixture were fixed to 10 mg/l reaction mixture solution
and 20°C, respectively. Open circles, open triangles, open squares,
closed circles and closed triangles indicate pH 7.0, pH 8.0, pH 9.0, pH
10.0, and pH 11.0, respectively.
30). These results suggest that EPS derived from PS88 is
capable of drug adsorption and that it is possible that DDS
functions as an EPS–gel–drug conjugate. Tegaful–EPS con-
jugate was rated as a weak conjugate compared with 5FU–
EPS conjugate. Tegaful was released easily as time elapsed.
To apply EPS for DDS, the release rate of a drug from EPS
is also important. As shown in Fig. 7, drug (5FU [Fig. 7a],
Tegaful [Fig. 7b]) release profile at different temperatures
was examined. The amount of 5FU release from the EPS–
drug conjugate increased with an increase in temperature
(Fig. 7a). The release of 5FU from the EPS–drug conjugate
was relatively repressed at lower temperatures (below 20°C),
thus thermosensitive drug release was observed from the
EPS–drug conjugate. Similar release profiles were obtained
J. BIOSCI. BIOENG.,
FIG. 6. Adsorption of Tegaful onto EPS gel under different pHs.
(a) EPS, Mg2+ and Tegaful in reaction mixture. (b) Mg2+ and Tegaful in
reaction mixture as control experiment. (c) Estimated adsorbed Tegaful
onto EPS gel. The initial Tegaful concentration and temperature in the
reaction mixture were fixed to 10 mg/l reaction mixture solution and
20°C, respectively. Symbols are the same as those in Fig. 5.
from Tegaful as shown in Fig. 7b. The mechanism underly-
ing the formation of the EPS–drug conjugate is still under
investigation. EPS is a gel that forms a mucus substance
(27, 28); therefore, the adsorption of a drug onto EPS might
mainly occur due to the entrapment of the drug by EPS gel
similar to chitosan because chitosan is a mucus substance
and it forms 5FU and Tegaful conjugates mainly by entrap-
ment (29, 30).
These results suggest the feasibility that this EPS from
PS88 is clinically applicable to thermo-chemotherapy. These
findings also suggest that EPS can be applied as pH-sensi-
tive biodegradable materials like chitosan. The advantage of
EPS derived from PS88 is mainly related to RNA. RNA
functions for the improvement of the immune function and
hepatic metabolism in humans (32), as described above. In
VOL. 100, 2005
FIG. 7. Effects of temperature on release profiles of (a) 5FU and
(b) Tegaful from EPS–drug conjugates in physiological saline solution.
EPS–drug conjugates (adsorbed drugs at pH 8.0 for 40 h) were resus-
pended in 10 ml of physiological saline buffer at 20–50°C and the
amounts of 5FU and Tegaful released in the supernatant from EPS were
measured according to Ohya’s method (29). Open circles, open tri-
angles, open squares and closed circles indicate 20°C, 30°C, 40°C and
50°C, respectively.
addition, EPS can be produced abundantly using relatively
low-cost fermentation technology (28). On the other hand,
chitosan is produced from natural materials such as crabs
and crayfish, which are limited resources, therefore, chito-
san production depends on the amount of these natural ma-
terials harvested.
Details of the mechanism of drug entrapment by EPS re-
main unclear, and further studies are needed to understand
the practical utilization of EPS as a drug carrier or adsor-
bent, including its biodegradability, stability and the struc-
ture of EPS.
Thus, various types of cell material of photosynthetic bac-
teria may be applicable in medical fields. As an example of
ALA applications, the bioproduction of ALA was only per-
formed from photosynthetic bacterial culture. More detailed
research studies are required to apply some components of
photosynthetic bacteria in the medical field.
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