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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

Inactive Publication Date: 2006-10-11
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The activity of ordinary lipase is reduced or completely inactivated under high salt concentration conditions, which limits its application
[0004] There is currently a lack of lipases that can tolerate high salt concentrations

Method used

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  • Halobacterium jilantaiense and application thereof
  • Halobacterium jilantaiense and application thereof
  • Halobacterium jilantaiense and application thereof

Examples

Experimental program
Comparison scheme
Effect test

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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PUM

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Abstract

This invention discloses a strain of Halobacterium jilantaiense and the application thereof. This strain is Halobacterium jilantaiense AC3 CGMCC No.1594, whose 16S rRNA encoding gene has the nucleotide sequence indicated by the No.1 sequence in the sequence table. The strain can produce lipase with tolerance to high salt concentration. The auxocyte, cell suspension, or the lipasse can catalyse the hydrolysis and synthesis of lipids in the range of 1.0-5.0M concentration of NaCl. And the lipase produced by this strain can catalyse triglyceride and other glycerides in high salt concentration solution or organic solvent.

Description

technical field [0001] The invention relates to a strain of halophilic bacillus gilantai and its application, in particular to the application of the strain of halophilic bacillus gilantai in the production of lipase. Background technique [0002] Lipase (lipase, EC 3.1.1.3) is widely distributed in nature and catalyzes the synthesis of triglyceride from fatty acid and glycerol and its reverse reaction. Lipase is a very important industrial enzyme, which is widely used in the processing and manufacturing of food, cosmetics, leather, paper, and washing products. Lipase catalyzes the hydrolysis and synthesis of water-insoluble triglycerides. The catalytic reaction is heterogeneous catalysis at the oil-water interface. The reaction rate is not only limited by the catalytic ability of the enzyme itself, but also depends on the size of the contact interface between the substrate and the enzyme. related. [0003] In many fields of application of lipase, catalytic conditions with...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N15/31C12N9/16C12P7/64
Inventor 周培瑾刘双江杨勇刘志培姜成英
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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