capped with a silicon plug; it was then autoclaved at 121
C for 15 min to sterilize
it. After having cooled to room temperature, 30 mL of the N-system, which was
cultured for 2–8 weeks, was added as a seed and cultured with an incubation
involving an L/D cycle ¼ 12 hr/12 hr, with an illuminance of 2400 lux (photosynthesis photon flux density ¼ 36 μmol/m
2 /s) at 25
C. The culture was stirred lightly
by hand once per day until culturing began 1–3 days later, after which time it was
allowed to rest. Subculturing was performed once every 2 months. A National (a.k.
a., Panasonic) FL20SS-W/18 was used as a source of light, and all fluorescent bulbs
were changed every 3 months or as needed based upon the illumination or a
photosynthesis photon flux density, measured every month. The conditions of
incubation of the microcosm (Figs. 3.1 and 3.2) (i.e., L/D ¼ 12 hr/12 hr, illuminance ¼ 2400 lux, and 25
C) as well as the appearance (i.e., the culturing N-system
in a 300 mL volume flask capped with a silicon plug) of the microcosm are shown in
Fig. 3.2. Measurement of production and respiration amount is shown in Fig. 3.3.
– Silicon plug: Matsuura Machinery, Catalog No. 6-343-16, Type No. C-40; ventilation amount ¼ 2000–4500 mL/min.
– Polypeptone: Wako Chemicals Co., Code No.394-00115 (500 g), 390-00117
(10 kg). Polypeptone (Nihon Pharmaceutical Co., Ltd.) is an ashy yellow dry
powder, which is purified after zymolysis of milk casein, and is equivalent to the
Trypticase Peptone and Casitone of BD Co., Ltd.
– Additional methods of Polypeptone depended upon each experiment (e.g., it was
added as a powder each time or an autoclaved stock solution of Polypeptone was
made and added aseptically each time, etc.).
Table 3.1 Taub Polypeptone medium
Stock solutions
Volume
A
Polypeptone*
2、3、4
2.0 g
/100 ml
10.0 ml
B
MgSO 4 ・7H 2 O
4.93 g
/200 ml
2.0 ml
C
K H 2 PO 4
2.72 g
/200 ml
2.0 ml
NaOH
0.56 g
D
CaCl 2 ・2H 2 O
7.35 g
/500 ml
40.0 ml
E
NaCl
2.92 g
/500 ml
60.0 ml
F
FeSO 4 ・7H 2 O
4.98 g
/200 ml
0.375 ml
Na 2 EDTA
2.72 g
G
H 3 BO 4
0.930 g
/500 ml
1.0 ml
ZnSO 4 ・7H 2 O
0.144 g
MnCl 2 ・4H 2 O
0.99 g
Na 2 MoO 4 ・2H 2 O
0.012 g
CuSO 4 ・5H 2 O
0.025 g
Co(NO 3 ) 2 6H 2 O
0.145 g
D.W.
Up to 2000 ml
Total nitrogen (T-N) ¼ 100 mg/L Â 14.5% ¼ 14.5 mg/L
Total phosphorus (T-P) ¼ 1.0(peptone) + 3.1(KH 2 PO 4 ) ¼ 4.1 mg/L
T-N/T-P ¼ 3.54
20
K. Shibata et al.
C for 15 min to sterilize
it. After having cooled to room temperature, 30 mL of the N-system, which was
cultured for 2–8 weeks, was added as a seed and cultured with an incubation
involving an L/D cycle ¼ 12 hr/12 hr, with an illuminance of 2400 lux (photosynthesis photon flux density ¼ 36 μmol/m
2 /s) at 25
C. The culture was stirred lightly
by hand once per day until culturing began 1–3 days later, after which time it was
allowed to rest. Subculturing was performed once every 2 months. A National (a.k.
a., Panasonic) FL20SS-W/18 was used as a source of light, and all fluorescent bulbs
were changed every 3 months or as needed based upon the illumination or a
photosynthesis photon flux density, measured every month. The conditions of
incubation of the microcosm (Figs. 3.1 and 3.2) (i.e., L/D ¼ 12 hr/12 hr, illuminance ¼ 2400 lux, and 25
C) as well as the appearance (i.e., the culturing N-system
in a 300 mL volume flask capped with a silicon plug) of the microcosm are shown in
Fig. 3.2. Measurement of production and respiration amount is shown in Fig. 3.3.
– Silicon plug: Matsuura Machinery, Catalog No. 6-343-16, Type No. C-40; ventilation amount ¼ 2000–4500 mL/min.
– Polypeptone: Wako Chemicals Co., Code No.394-00115 (500 g), 390-00117
(10 kg). Polypeptone (Nihon Pharmaceutical Co., Ltd.) is an ashy yellow dry
powder, which is purified after zymolysis of milk casein, and is equivalent to the
Trypticase Peptone and Casitone of BD Co., Ltd.
– Additional methods of Polypeptone depended upon each experiment (e.g., it was
added as a powder each time or an autoclaved stock solution of Polypeptone was
made and added aseptically each time, etc.).
Table 3.1 Taub Polypeptone medium
Stock solutions
Volume
A
Polypeptone*
2、3、4
2.0 g
/100 ml
10.0 ml
B
MgSO 4 ・7H 2 O
4.93 g
/200 ml
2.0 ml
C
K H 2 PO 4
2.72 g
/200 ml
2.0 ml
NaOH
0.56 g
D
CaCl 2 ・2H 2 O
7.35 g
/500 ml
40.0 ml
E
NaCl
2.92 g
/500 ml
60.0 ml
F
FeSO 4 ・7H 2 O
4.98 g
/200 ml
0.375 ml
Na 2 EDTA
2.72 g
G
H 3 BO 4
0.930 g
/500 ml
1.0 ml
ZnSO 4 ・7H 2 O
0.144 g
MnCl 2 ・4H 2 O
0.99 g
Na 2 MoO 4 ・2H 2 O
0.012 g
CuSO 4 ・5H 2 O
0.025 g
Co(NO 3 ) 2 6H 2 O
0.145 g
D.W.
Up to 2000 ml
Total nitrogen (T-N) ¼ 100 mg/L Â 14.5% ¼ 14.5 mg/L
Total phosphorus (T-P) ¼ 1.0(peptone) + 3.1(KH 2 PO 4 ) ¼ 4.1 mg/L
T-N/T-P ¼ 3.54
20
K. Shibata et al.