Method for producing l-glutamine by fermentation and l-glutamine producing bacterium
a technology of l-glutamine and bacterium, which is applied in the direction of enzymology, ligases, transferases, etc., can solve the problems of insufficient l-glutamine yield, low production ability of wild strains per se for l-amino acid production, and insufficient l-glutamine fermentation. , to achieve the effect of suppressing the by-product of l-glutamine production and efficient production
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Benefits of technology
Problems solved by technology
Method used
Examples
example 1
Evaluation of GS Gene-amplified Strain
(1) Cloning of glnA Gene of Coryneform Bacterium
[0158] The glnA sequence of Corynebacterium glutamicum had been already clarified (FEMS Microbiology Letters, 81-88, (154) 1997). Based on the reported nucleotide sequence, the primers shown in Sequence Listing as SEQ ID NOS: 4 and 5 were synthesized, and a glnA fragment was amplified by the PCR method using chromosome DNA of Brevibacterium flavum ATCC 14067 strain as a template.
[0159] The chromosomal DNA of Brevibacterium flavum ATCC 14067 strain was prepared by using Bacterial Genome DNA Purification Kit (Advanced Genetic Technologies Corp.). PCR was performed for 30 cycles each consisting of reactions at 94° C. for 30 seconds for denaturation, at 55° C. for 15 seconds for annealing and 72° C. for 2 minutes for extension by using Pyrobest DNA Polymerase (Takara Shuzo).
[0160] The produced PCR product was purified in a conventional manner, digested with a restriction enzyme SalI, ligated with ...
example 2
Evaluation of GS Adenylylation Site-modified Strain
(1) Construction of Adenylylation Site-modified Plasmid
[0167] The adenylylation site of glnA gene product of coryneform bacteria had been already clarified (FEMS Microbiology Letters, 303-310, (173) 1999). Therefore, an adenylylation site-modified strain was obtained by replacing the glnA gene on the chromosome with a glnA gene of which adenylylation site was modified. Specific procedures will be described below.
[0168] First, PCR was performed by using chromosome DNA of Brevibacterium flavum ATCC 14067 strain as a template and the synthetic DNAs shown in Sequence Listing as SEQ ID NOS: 6 and 7 as primers to obtain an amplification product for the N-terminus side of the glnA gene. Separately, in order to obtain an amplification product for the C-terminus side of the glnA gene, PCR was performed by using chromosome DNA of Brevibacterium flavum ATCC 14067 strain as a template and the synthetic DNAs shown in Sequence Listing as SEQ ...
example 3
Evaluation of GDH Gene-amplified Strain
(1) Construction of gdh-amplified Strain and Evaluation of Culture
[0173] Construction of a plasmid pGDH into which the gdh gene of coryneform bacteria was cloned was performed as follows. First, chromosome DNA of Brevibacterium lactofermentum ATCC 13869 strain was extracted, and PCR was performed by using the chromosome DNA as a template and the synthetic DNAs shown in Sequence Listing as SEQ ID NOS: 12 and 13 as primers. The obtained DNA fragment was blunt-ended and inserted into the SmaI site of pHSG399 (Takara Shuzo). This plasmid was designated as pHSG399GDH.
[0174] Then, a replication origin derived from the plasmid pHM1519 (Agric. Biol. Chem., 48, 2901-2903 (1984)) that could autonomously replicate in coryneform bacteria was introduced into the SalI site of pHSG399GDH. Specifically, the aforementioned pHK4 was digested with restriction enzymes BamHI and KpnI to obtain a gene fragment containing the replication origin, and the obtained ...
PUM
Property | Measurement | Unit |
---|---|---|
Temperature | aaaaa | aaaaa |
Fraction | aaaaa | aaaaa |
Fraction | 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