A strain of Geobacillus and its application in the production of 2,3-butanediol and acetoin by high-temperature fermentation
A Geobacillus, high-temperature fermentation technology, applied in the biological field
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Embodiment 1
[0023] Example 1: Strain enrichment screening and isolation and purification
[0024] 1.1 Preparation of enrichment medium
[0025] Collect produced water samples from Shengli Oilfield, add 20g of glucose, 20g of peptone, and 10g of yeast powder per liter of water sample. Adjust the pH to 7.0 to obtain an enriched medium.
[0026] 1.2 Strain enrichment
[0027] The above-mentioned enriched medium was divided into shake flasks, rapidly heated to 100° C., and then rapidly transferred to a water bath shaker at 60° C. for 2 d with reciprocating shaking (140 rpm). In this step, short-time sterilization at 100°C can kill most of the heat-labile interfering bacteria, and 60°C shaker culture can further amplify thermophilic bacteria and eliminate common bacillus and anaerobic bacteria.
[0028] 1.3 Isolation and purification of strains
[0029] Prepare solid seed culture medium, each liter contains: 20g glucose, 20g peptone, 10g yeast powder, 17g agar powder. Adjust the pH of the...
Embodiment 2
[0032] Embodiment 2: strain identification and preservation
[0033] 2.1 Observation of morphological characteristics
[0034] The colony of the target bacterial strain obtained by the above-mentioned Example 1 is white, with wrinkles on the surface and uneven edges. Gram stain was positive, producing spores. The cells are rod-shaped, 0.9-1.8 μm long and 0.4-0.5 μm in diameter (figure).
[0035] 2.2 Identification and preservation of strain 16S rDNA
[0036] Using bacterial 16S rDNA universal primers 27F and 1492R as amplification primers, the 16S rDNA fragment of the target strain obtained in the above 1.3 experiment was amplified by PCR method, and after electrophoresis detection, it was sent to Shanghai Sangon Bioengineering Technology Service Co., Ltd. Sequencing results, and using the BLAST program of the National Center for Biotechnology Information (NCBI) for nucleotide homology comparison between the 16S rDNA sequence of the bacteria and the sequence already include...
Embodiment 3
[0042] Embodiment 3: determine optimal culture temperature
[0043] Follow the conventional method for determining the optimum culture temperature in microbiology experiments. Briefly, the Geobacillus XT15 described in Example 2 was inoculated in a liquid seed medium and cultured on a shaker under different temperature conditions. The changes in the OD value of the culture were measured with a spectrophotometer, and the average cell proliferation rate was calculated. Each liter of the liquid seed culture medium contains: 20g of glucose, 20g of peptone, and 10g of yeast powder.
[0044] As determined by the above experiments, the strain Geobacillus XT15 of the present invention can grow in the range of 28-65°C; the proliferation is fastest in the range of 45-55°C, and the number of cells doubles about every 2h. In the range of 45-55°C, the difference in proliferation rate is very small. In order to meet the needs of high-temperature fermentation, 55°C is selected as the optim...
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Abstract
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