A kind of new Halomonas and method for producing tetrahydropyrimidine
A technology of Halomonas and tetrahydropyrimidine, applied in the field of microorganisms, can solve the problems of unsuitability for large-scale industrial production, less tetrahydropyrimidine, corrosion of fermentation equipment and the like
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0027] Example 1: Obtaining Halomonas sp. HS-2255 (CGMCCNO.6248)
[0028] Take a 1g sample of sea mud from the coast of Taizhou, Zhejiang, and serially dilute it with sterile water, spread it on an LB plate containing different concentrations of NaCl (30-100g / L), incubate at 30°C, pick a single colony after the colony grows, and continue to separate Purify and screen until pure bacteria are obtained, cultivate them in a fermentation medium with a lower NaCl concentration (not exceeding 100g / L, optimally 75g / L), and detect the ectoine production of the strains by high-pressure liquid chromatography. Select mutant strains obtained through mutagenesis screening against high-dose metabolic end products and structural analogs, carry out shake flask screening, measure and evaluate the ability and genetic stability of ectoine production, and obtain an excellent strain with significantly improved yield, The mutant strain for mutagenesis screening and resistance screening, and the stra...
Embodiment 2
[0029] Example 2: 16S rDNA amplification, sequencing and sequence alignment of Halomonas sp. HS-2255 (CGMCC NO.6248).
[0030] Collect the fresh bacteria of the strain to be tested, extract the total DNA template by the Pitcher method improved by the solution bacterial enzyme method, and use the universal primers (27F and 1495R) to amplify the 16SrDNA gene. After the PCR product is detected and purified, it is directly sequenced and sequenced. Performed by Shanghai Sangon Bioengineering Technology Co., Ltd.
[0031] The primer sequences are as follows:
[0032] a) 27F:5'-GAGAGTTTGATCCTGGCTCAG-3'(E.coli7-27)
[0033] b) 1495R: 5'-CTACGGCTACCTTGTTACGA-3' (E.coli1495-1514)
[0034] The 50μl PCR reaction system is as follows:
[0035] wxya 2 o
35μl
25 μM Primer 27F
1μl
25 μM Primer 1495R
1μl
10×Poly.Buffer
5μl
2.5mM dNTPs
4μl
25mM MgCl 2
3μl
DNA template
0.5ul
0.5μl
...
Embodiment 3
[0038] Example 3: Culture method of Halomonas sp. HS-2255 (CGMCCNO.6248)
[0039] Insert the strain of the present invention into the seed culture medium, the composition of the seed culture medium is: fish peptone 20g / L, glucose 20g / L, yeast extract 20g / L, NaCl 40g / L, pH7.5, culture temperature is 28-35°C, The shaker speed is 180-250r / min, and the culture time is 24-48h, until the seed solution OD 600 The absorption value is 0.1-1.0 (with water as a reference, the seed liquid is diluted 30 times).
PUM
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