Marine low-temperature alkaline lipase Bohai Sea-9145 crystal and preparation method thereof
A lipase, low-temperature technology, applied in the field of marine microorganisms, can solve the problems of undiscovered, and achieve the effects of short fermentation period, high low-temperature catalytic activity, and wide source of raw materials
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0028] Example 1——Preparation of marine low-temperature alkaline lipase Bohai Sea-9145 crystals
[0029] 1) Preparation of crystallization (growth) buffer: Prepare a 0.1M sodium acetate buffer with a pH of 4.6, which contains 20% by mass of ammonium sulfate as a precipitant.
[0030] 2) Preparation of marine low-temperature alkaline lipase Bohai Sea-9145 aqueous solution
[0031] The marine low-temperature alkaline lipase Bohai Sea-9145 with an enzyme activity of 4000U / mL was prepared as a 10mg / ml aqueous solution.
[0032] 3) Preparation of crystallization solution:
[0033] 1 μL of crystallization buffer prepared in the above step 1, 1 μL of 20 mg / ml marine yeast low-temperature alkaline lipase Bohai Sea-9145 aqueous solution; using the hanging drop method, place the above crystallization solution on a plastic cover glass, and then drop the crystallization solution Put it upside down on a 24-well crystallization plate, seal the gap in the middle with Vaseline, culture the ...
Embodiment 2、3
[0034] Embodiment 2, 3—Preparation of marine low-temperature alkaline lipase Bohai Sea-9145 crystal
[0035] The preparation procedure of embodiment 2,3 is identical with embodiment 1, and difference is: wherein prepare respectively 0.2M, 0.3M, pH is the sodium acetate buffer solution of 4.6, and the mass percent content containing precipitating agent ammonium sulfate is respectively 25% 30% ; The prepared marine low-temperature alkaline lipase Bohai Sea-9145 aqueous solution was 15, 20mg / ml respectively.
Embodiment 4
[0036] Example 4——Structural analysis of marine low-temperature alkaline lipase Bohai Sea-9145 crystal
[0037] First, use software such as HKL2000 to conduct X-ray diffraction on the crystal of marine low-temperature alkaline lipase Bohai Sea-9145, collect the obtained diffraction data for processing, and obtain a complete data file; secondly, use software such as Phaser and Molrep in the CCP4 program package, Using molecular replacement (MR, Molecular Replacement) method, Yarrowia lipolytica lipase (PDBid: 3O0D), thermophilic (Humicola) humic lipase (PDBid: 1DT3) and fungal lipase (PDBid: 1TIA) were used as Search the model and obtain the fine three-dimensional structure of marine low-temperature alkaline lipase Bohai Sea-9145 ( figure 2 , 3 ).
PUM
Property | Measurement | Unit |
---|---|---|
molecular weight | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com