Halobacterium jilantaiense and application thereof
A technology of halophilic bacteria and lipase, applied to halophilic bacteria gilante and its application fields, can solve the problems of application limitation, activity reduction, inactivation and the like
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Embodiment 1
[0019] Example 1. Isolation and Identification of Halobacterium jilantaiense AC3 CGMCC No.1594
[0020] 1. The acquisition and biological characteristics of Halobacterium jilantaiense AC3 CGMCC No.1594
[0021] Show through culture experiment, this bacterium has following characteristics: (1) bacterium colony characteristic: on milk salt plate (150mL of skimmed milk, inorganic salt solution (MgSO 4 ·7H 2 O 3g, NaCl 60g, KNO 3 0.6g, 0.5mg ferric citrate) 30mL, agar solution (Casamino acid 1.5g, glycerin 3g, agar 4.5g) 120ml) for 7 days, the colony is round, the diameter of the colony is 0.5-2.1mm, the surface is smooth and the edges are neat , raised, cinnamon red. (2) Halobacterium jilantaiense AC3 CGMCC No.1594 strain grows strictly aerobically; the suitable pH range for growth is 6-9, and the optimum growth pH range is 7.5-8.5; the growth temperature range is 35-45°C, The optimum temperature is 40°C; the cells are short rod-shaped; Gram staining is negative; the cell si...
Embodiment 2
[0026] Example 2, the growth of Halobacterium jilantaiense AC3 CGMCC No.1594 under different NaCl concentration conditions and the substrate scope of the lipase it produces
[0027] 1. Growth of Halobacterium jilantaiense AC3 CGMCC No.1594 under different NaCl concentrations
[0028] Halobacterium jilantaiense (Halobacterium jilantaiense) AC3 CGMCC No.1594 was respectively inoculated in 0.75g / 100ml casamino acid (Casamino acid, Difco), 1g / 100ml yeast extract (Yeast extract, Oxoid), 0.3 g / 100ml (0.068M) trisodium citrate trihydrate, magnesium sulfate heptahydrate 2.0g / 100ml (0.081M), potassium chloride 0.2g / 100ml (0.0268M), ferrous sulfate heptahydrate 0.05g / 100ml (0.00018 M), sodium chloride concentrations are 6g / 100ml (1.03M), 8g / 100ml (1.36M), 10g / 100ml (1.7M), 12g / 100ml (2.05M), 14g / 100ml (2.40M), 16g / 100ml(2.74M), 18g / 100ml(3.08M), 20g / 100ml(3.42M), 22g / 100ml(3.76M), 24g / 100ml(4.11M), 26g / 100ml(4.45M), 28g / 100ml (4.80M), 30g / 100ml (5.13M) and 1000ml of distilled water i...
Embodiment 3
[0031] Example 3, Purification of lipase produced by Halobacterium jilantaiense AC3 CGMCC No.1594 and determination of enzyme activity:
[0032] 1. Purification of lipase produced by Halobacterium jilantaiense AC3 CGMCC No.1594
[0033] Halobacterium jilantaiense AC3 CGMCC No.1594 was cultured in the following CM liquid medium: Casamino acid (Difco) 7.5g, Yeast extract (Oxoid) 10.0g, MgSO 4 ·7H 2 O 20.0g, Trisodium citrate 3.0g, KCl 2.0g, FeSO 4 ·7H 2 O 0.05g, NaCl 200g, distilled water 1000ml, adjust the pH to 7.0-7.4 with 1M NaOH, and sterilize at 121°C for 30 minutes. Cultivate at 40°C, light and ventilate (the amount of liquid filled is 1 / 5 of the volume of the shake flask, the radius of rotation is 35 mm, and 200 rpm) for 5-7 days. Centrifuge the cultured bacterial solution at 8,000-12,000 rpm for 5-10 minutes, add 20-30% volume of acetone to the supernatant, centrifuge at 8,000-12,000 rpm for 5-10 minutes, and store the collected precipitate on ice Dissolve in the p...
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