Desulfotomaculum acetoxidans Medium

[所属分类:培养基配方] [发布时间:2020-6-8] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Desulfotomaculum acetoxidans Medium

培养基配方
Composition per 1011.0mL:
Solution A..................................................................1.0L
Solution B................................................................10.0mL
Vitamin solution...........................................................1.0mL
pH 7.0 ± 0.2 at 25°C
Solution A:
Composition per liter:
NaHCO3 ......................................................................4.5g
Na2SO4 .....................................................................2.84g
Sodium acetate...............................................................1.4g
Sodium butyrate..............................................................1.4g
NaCl........................................................................1.17g
Yeast extract................................................................1.0g
MgCl2 ·6H2O ................................................................0.4g
KCl..........................................................................0.3g
NH4Cl ......................................................................0.27g
KH2PO4 ......................................................................0.2g
CaCl2 ·2H2O ...............................................................0.15g
Resazurin ..................................................................0.5mg
Trace elements solution .....................................................1.0mL
Preparation of Solution A: Add components, except NaHC3 and
vitamin solution, and bring volume to 1.0L. Mix thoroughly. Gently
heat and bring to boiling. Continue boiling for 3–4 min. Allow to cool
to room temperature while gassing under O2 -free 80% N2 + 20% CO2 .
Add NaHCO3 and continue gassing with O2 -free 80% N2 + 20% CO2
until pH reaches 6.9–7.1. Seal the flask under 80% N2 + 20% CO2 . Au-
toclave for 15 min at 15 psi pressure–121°C.
Trace Elements Solution:
Composition per liter:
FeCl2 ·4H2O..................................................................1.5g
CoCl2 ·6H2O ..............................................................120.0mg
MnCl2 ·4H2O...............................................................100.0mg
ZnCl2 ......................................................................68.0mg
H3BO3 ......................................................................62.0mg
Na2MoO4 ·2H2O .............................................................24.0mg
NiCl2 ·6H2O................................................................24.0mg
CuCl2 ·2H2O ...............................................................17.0mg
HCl (25% solution)..........................................................10.0mL
Preparation of Trace Elements Solution:  Add FeCl2 ·4H2O to
10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized wa-
ter and bring volume to 1.0L. Add remaining components. Mix thor-
oughly. Gas under 100% N2 .
Vitamin Solution:
Composition per 100.0mL:
Thiamine·HCl ............................................................10.0mg
p-Aminobenzoic acid........................................................4.0mg
D (+)-Biotin...............................................................1.0mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize. Gas under 100% N2 .
Solution B:
Composition per 10.0mL:
Na2S·9H2O.................................................................0.36g
Preparation of Solution B:  Add Na2S·9H2O to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Gas under 100%
N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: To 1.0L of sterile solution A, add 10.0mL
of sterile solution B and 1.0mL of sterile vitamin solution. Mix thor-
oughly.
Use:  For the cultivation and maintenance of Desulfotomaculum ace-
toxidans.