Artificial Organic Lake Peptone Medium

[所属分类:培养基配方] [发布时间:2019-11-11] [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

Artificial Organic Lake Peptone Medium
Composition per 1001.0mL:
NaCl............................................................................................30.0g
MgSO4 ·7H2O................................................................................9.5g
KCl................................................................................................5.0g
Peptone..........................................................................................5.0g
Yeast extract..................................................................................1.0g
CaCl2 ·2H2O...................................................................................0.2g
KNO3 ............................................................................................0.1g
(NH4)2SO4 .....................................................................................0.1g
Modified Hutner’s basal salts ..................................................20.0mL
Phosphate supplement..............................................................20.0mL
Artificial organic lake vitamin solution.....................................1.0mL
pH 7.3 ± 0.2 at 25°C
Modified Hutner’s Basal Salts:
Composition per liter:
MgSO4 ·7H2O..............................................................................29.7g
Nitrilotriacetic acid.....................................................................10.0g
CaCl2 ·2H2O................................................................................3.34g
FeSO4 ·7H2O ............................................................................99.0mg
Ammonium molybdate............................................................9.25mg
Metals “44”..............................................................................50.0mL
Preparation of Modified Hutner’s Basal Salts:  Dissolve the
nitrilotracetic acid first and neutralize the solution with KOH. Add the
other components and adjust the pH to 7.2 with KOH or H 2 SO 4 . There
may be a slight precipitate. Store at 5°C.
Metals “44”
Composition per liter:
ZnSO 4 ·7H2O.................................................................................1.1g
FeSO4 ·7H2O .................................................................................0.5g
CuSO4 ·5H2O...............................................................................0.04g
EDTA..........................................................................................0.25g
MnSO4 ·7H2O............................................................................0.154g
Co(NO3)2 ·6H2O........................................................................0.025g
Na2B4O7 ·10H2O.......................................................................0.018g
Preparation of Metals “44”:  Add components to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. Autoclave
for 15 min at 15 psi pressure–121°C. Add aseptically to sterile modi-
fied Hutner’s basal salts solution.
Phosphate Supplement:
Composition per liter:
K2HPO4 .........................................................................................2.5g
KH2PO4 .........................................................................................2.5g
Preparation of Phosphate Supplement: Add components to dis-
tilled/deionized water and bring volume to 20.0mL. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C.
Artificial Organic Lake Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Nicotinamide..............................................................................5.0mg
Riboflavin ..................................................................................5.0mg
Thiamine·HCl ............................................................................5.0mg
Biotin.........................................................................................2.0mg
Folic acid ...................................................................................2.0mg
Cyanocobalamin........................................................................0.1mg
Preparation of Artificial Organic Lake Vitamin Solution:
Add components to distilled/deionized water and bring volume to
1.0L. Mix thoroughly. Filter sterilize. Store at 5°C.
Preparation of Medium: Add components, except modified Hut-
ner’s basal salts solution, phosphate supplement solution, and vitamin
solution, to distilled/deionized water and bring volume to 959.0mL.
Mix thoroughly. Bring pH to 7.3. Autoclave for 15 min at 15 psi pres-
sure–121°C. Cool to 50°C. Aseptically add 20.0mL of sterile modified
Hutner’s basal salts solution, 20.0mL of sterile phosphate supplement
solution, and 1.0mL of sterile artificial organic lake vitamin solution.
Mix thoroughly. Aseptically distribute into sterile tubes or flasks.
Use: For the cultivation of Halomonas meridiana.