Halocella cellulolytica Medium

[所属分类:培养基配方] [发布时间:2021-3-26] [发布人: ] [阅读次数:] [返回]

Halocella cellulolytica Medium

培养基配方

Composition per liter:
NaCl..........................................................................................150.0g
Cellobiose or microcrystalline cellulose.......................................5.0g
Yeast extract..................................................................................2.0g
MgCl 2 ·6H 2 O .................................................................................3.6g
CaCl 2 ...........................................................................................0.33g
KCl..............................................................................................0.33g
KH 2 PO 4 .......................................................................................0.33g
NH 4 Cl .........................................................................................0.33g
Resazurin...................................................................................2.0mg
NaHCO 3  solution.....................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Wolfe’s vitamin solution..........................................................10.0mL
Trace elements solution SL-10..................................................1.0mL
pH 7.0 ± 0.2 at 25°C
NaHCO 3 Solution:
Composition per 10.0mL:
NaHCO 3 ........................................................................................2.5g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Sparge with 100% N 2 .
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.5g
Preparation of Na 2 S·9H 2 O Solution:  Add Na 2 S·9H 2 O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
p-Aminobenzoic acid.................................................................5.0mg
Lipoic acid.................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
Riboflavin..................................................................................5.0mg
Thiamine·HCl............................................................................5.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Biotin.........................................................................................2.0mg
Folic acid ...................................................................................2.0mg

Vitamin B 12 ................................................................................0.1mg

Preparation of Wolfe’s Vitamin Solution:  Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize.
Trace Elements Solution SL-10:
Composition per liter:
FeCl 2 ·4H 2 O...................................................................................1.5g
CoCl 2 ·6H 2 O ...........................................................................190.0mg
MnCl 2 ·4H 2 O...........................................................................100.0mg
ZnCl 2 ........................................................................................70.0mg
Na 2 MoO 4 ·2H 2 O .......................................................................36.0mg
NiCl 2 ·6H 2 O..............................................................................24.0mg
H 3 BO 3 ........................................................................................6.0mg
CuCl 2 ·2H 2 O ...............................................................................2.0mg
HCl (25% solution)..................................................................10.0mL
Preparation of Trace Elements Solution SL-10: Add FeCl 2 ·4H 2 O to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized water and bring volume to 1.0L. Add remaining components. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium:  Prepare and dispense medium under 80% N 2 + 20% CO 2 . Add components, except NaHCO 3  solution, Na 2 S·9H 2 O solution, and Wolfe’s vitamin solution, to distilled/deionized water and bring volume to 970.0mL. Mix thoroughly. Gently heat and bring to boiling. Cool to room temperature while sparging with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically add 10.0mL of sterile NaHCO 3 solution, 10.0mL of sterile Na 2 S·9H 2 O solution, and 10.0mL of sterile Wolfe’s vitamin solution. Mix thoroughly. Check that final pH is 7.0.
Use: For the cultivation of Halocella cellulolytica.