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Halobacterium jilantaiense and application thereof

A technology of halophilic bacteria and high salt, applied to halophilic bacteria of gilante and its application fields, can solve the problems of lack of lipase, application limitation, activity reduction and the like

Inactive Publication Date: 2008-12-24
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 AC3CGMCC No. 1594

[0020] 1. Acquisition of Halobacterium jilantaiense AC3CGMCC No. 1594 and its biological characteristics

[0021] The culture experiment shows that the bacteria has the following characteristics: (1) Colony characteristics: on a 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, glycerol 3g, agar 4.5g) 120ml), the colonies after 7 days of cultivation are round, the colony diameter is 0.5-2.1mm, the surface is smooth, and the edges are neat , Raised, laurel red. (2) Halobacterium jilantaiense (Halobacterium jilantaiense) AC3CGMCC No. 1594 grows strictly aerobic; the suitable pH range for growth is 6-9, the optimal pH range for growth is 7.5-8.5; the growth temperature range is 35-45°C, the most The suitable temperature is 40℃; the cells are short rod-shaped; Gram staini...

Embodiment 2

[0026] Example 2: Growth of Halobacterium jilantaiense AC3CGMCC No. 1594 under different NaCl concentration conditions and the range of substrates of lipase produced

[0027] 1. Growth of Halobacterium jilantaiense AC3CGMCC No. 1594 under different NaCl concentrations

[0028] Halobacterium jilantaiense (Halobacterium jilantaiense) AC3CGMCC No. 1594 was respectively inoculated into 0.75g / 100ml Casamino acid (Difco), 1g / 100ml Yeast extract (Oxoid), 0.3g / 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 (0.00018M) ), the concentration of sodium chloride is 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 in a liquid medium (adjusted to pH 7...

Embodiment 3

[0031] Example 3. Purification of lipase produced by Halobacterium jilantaiense AC3CGMCC No. 1594 and determination of enzyme activity:

[0032] 1. Purification of lipase produced by Halobacterium jilantaiense AC3CGMCC No. 1594

[0033] The Halobacterium jilantaiense AC3CGMCC No. 1594 was cultured in the following CM liquid medium: Casamino acid (Difco) 7.5 g, Yeast extract (Oxoid) 10.0 g, MgSO 4 ·7H 2 O 20.0g, Trisodium citrate 3.0g, KCl 2.0g, FeSO 4 ·7H 2 O 0.05g, NaCl 200g, 1000ml distilled water, adjust the pH to 7.0-7.4 with 1M NaOH, sterilize at 121 degrees Celsius for 30 minutes. Cultivation temperature is 40°C, light and aeration (filling liquid volume is 1 / 5 of shake flask volume, 35mm rotation radius, 200 revolutions / min) for 5-7 days. Put the cultured bacterial solution at 8,000-12,000 rpm for 5-10 minutes, add 20-30% volume of acetone to the supernatant, and 8,000-12,000 rpm for 5-10 minutes. The collected precipitate is placed on ice. Dissolve it in the pre-cooled 0.0...

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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 present invention relates to a strain of halophilus gilantea and its application, and particularly relates to the application of the strain of halophilus gilantea in the production of lipase. Background technique [0002] Lipase (EC 3.1.1.3) is widely distributed in nature and catalyzes the synthesis of triglycerides and the reverse reaction of fatty acids and glycerol. Lipase is a very important industrial enzyme, which is widely used in the processing and manufacturing of food, cosmetics, leather, paper, and detergents. Lipase catalyzes the hydrolysis and synthesis of non-water-soluble triglycerides. The catalytic reaction is a heterogeneous catalysis that occurs at the oil-water interface. The reaction rate is not only limited by the catalytic ability of the enzyme itself, but also by the size of the contact interface between the substrate and the enzyme. related. [0003] In many areas where lipases are applied, catalytic conditions with higher salt...

Claims

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Application Information

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Patent Type & Authority Patents(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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