Bacillus amyloliquefaciens and method for preparing Maotai-flavor sweet tobacco by using bacillus amyloliquefaciens
A technology for dissolving starch spores and bacilli, applied in the field of microorganisms, can solve problems such as the preparation and application of unconcerned strains in sauce-flavored fermentation broth, and achieve the effects of abundant raw materials, low production cost and little environmental pollution
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0052] Screening and isolation of embodiment 1 bacterial strain
[0053] S1: Weigh 10 g of high-temperature Daqu after grinding, add it to a conical flask filled with 90 mL of sterile normal saline, shake and culture at 37 °C and 180 r / min for 1 h, and place the prepared bacterial suspension in a water bath at 80 °C for 30 min .
[0054] S2: configuration 10 -3 、10 -4 、10 -5 、10 -6 、10 -7 Dilute the gradient, take 0.1mL respectively and spread it on the LB solid medium, and culture it upside down at 37°C for 24h.
[0055] S3: Select a single colony that grows well on the medium and has obvious morphological characteristics of the colony, streak the plate on the LB solid medium, and culture it upside down at 37°C for 24 hours. The numbered single colonies were inserted into the slant medium, cultured at 37°C for 24 hours, and stored at 4°C for later use.
[0056] For the specific ratio of the medium, see LB solid medium and slant medium in Table 1.
Embodiment 2
[0057] Embodiment 2 produces tetramethylpyrazine bacterial strain screening
[0058] The tetramethylpyrazine standard substance was made into different concentration gradients to make a standard curve. figure 1 Be the regression equation of tetramethylpyrazine standard curve: y=1.0412x-1.6985, R 2 = 0.9979.
[0059] About 103 suspected bacillus strains obtained by screening were inserted into 250ml shake flasks equipped with 50ml fermentation medium (using glucose as substrate) respectively, and were shaken and cultivated for 72h at 37°C and 180r / min. After the fermentation, the fermented liquid was diluted and extracted with 70% methanol, centrifuged at 10000r / min for 10min, and the supernatant was passed through a 0.22um organic filter membrane. Quantitative analysis of tetramethylpyrazine in the supernatant by HPLC detection. The fermentation broth was centrifuged at 5000r / min for 10min, the supernatant was extracted with 95% ethanol, and the components in the supernata...
Embodiment 3
[0066] Classification and Identification of Example 3 Bacterial Strains
[0067] The preserved strains were activated in LB medium, and 2ml of the bacterial liquid was centrifuged at 10,000r / min for 10min. After collecting the bacterial cells, the bacterial genome was extracted and amplified by PCR with bacterial universal primers 1492R and 27F. PCR reaction system (25 μL): plus and downstream primers 1.0 μL each, Mix 12.5 μL, DNA template 1.0 μL, ddH 2 O 9.5 μL. PCR program: pre-denaturation at 94°C for 10 minutes, and 30 cycles on this basis, the conditions are as follows: denaturation at 95°C for 30 seconds, annealing at 55°C for 50 seconds, extension at 72°C for 1.5 minutes, and finally extension at 72°C for 10 minutes. After the PCR, 1 μL of DNA was taken and poured into 1% agarose gel wells sequentially, and electrophoresed at 120V for 20 minutes. After the target band was determined, it was sent to Shanghai Bioengineering Co., Ltd. for sequencing. The BLAST compariso...
PUM
Login to view more Abstract
Description
Claims
Application Information
Login to view more - 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.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap