Bacillus amyloliquefaciens and applications of Bacillus amyloliquefaciens in co-production of bacterial cellulose and gamma-polyglutamic acid
A technology for dissolving starch spores and bacterial cellulose, applied in the field of fermentation engineering, can solve the problems of high production cost and low utilization rate of raw materials, and achieve the effects of low production cost, simple operation, and convenient and simple operation.
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Embodiment 1
[0037] Example 1: Identification of the genus of the strain NX-2S and the identification of the fermentation product γ-PGA and bacterial cellulose.
[0038] Genomic DNA of bacterial strain NX-2S was extracted using a bacterial genomic DNA extraction kit, and the 16S rDNA sequence was amplified by PCR with upstream primer 27F and downstream primer 1492R as follows: figure 1 As shown, PCR was purified and sent to Nanjing GenScript for sequencing. The sequencing results were compared with the known 16S rDNA sequences in the GenBank database by BLAST. The results showed that the strain NX-2S reached 100% homology with Bacillus amyloliquefaciens ZH1. According to the results of strain morphology observation and physiological and biochemical experiment analysis, it is determined that the present invention uses Bacillus amyloliquefaciens, specifically Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) NX-2S.
[0039] Take Bacillus amyloliquefaciens (Bacillus amyloliquefaciens)...
Embodiment 2
[0043] Example 2: Static fermentation of Bacillus amyloliquefaciens NX-2S in a 7.5L tank to produce γ-PGA and bacterial cellulose.
[0044] Seed medium: glucose 10g / L, yeast extract 3g / L, peptone 10g / L, MgSO 4 ·7H 2 O 0.5g / L, use NaOH solution to adjust the pH to 6.8.
[0045] Fermentation medium: glucose 60g / L, citric acid 1g / L, beef extract 10g / L, yeast extract 5g / L, Na 2 HPO 4 12H 2 O 5g / L, ethanol 6% (v / v), using Ca(OH) 2 The solution was adjusted to pH 6.8.
[0046] Cultivate Bacillus amyloliquefaciens NX-2S (CCTCC M 2016346) in the seed medium, 30°C, 200r / min shaker conditions for 14h, and inoculate the seed solution at a seed volume of 5% (v / v) in Cultivate in a fermenter pre-installed with 3.5L sterilized fermentation medium, culture conditions: static culture at 30°C, air ratio controlled at 1.2VVM, open pH automatic control device during fermentation, use HCl or Ca(OH ) 2 Control the pH value of the fermentation broth between 6.5-8.0. After 5 days of ferment...
Embodiment 3
[0049] Example 3: 7.5L tank statically fermented Bacillus amyloliquefaciens NX-2S to produce γ-PGA and bacterial cellulose.
[0050] Seed medium: glucose 10g / L, yeast extract 3g / L, peptone 10g / L, MgSO 4 ·7H 2 O 0.5g / L, use NaOH solution to adjust the pH to 6.8.
[0051] Fermentation medium: inulin crude liquid 40g / L, sucrose 10g / L, glucose 10g / L, citric acid 2g / L, peptone 10g / L, (NH 4 ) 2 SO 4 5g / L, Na 2 HPO 4 12H 2 O 6g / L, ethanol 3% (v / v), using Ca(OH) 2 The solution was adjusted to pH 6.8.
[0052] Cultivate Bacillus amyloliquefaciens NX-2S (CCTCC M 2016346) in the seed culture medium, 32°C, 200r / min shaker conditions for 14h, and inoculate the seed solution at a seed volume of 6% (v / v) in Cultivate in a fermenter pre-installed with 3.5L sterilized fermentation medium, culture conditions: static culture at 32°C, air ratio controlled at 1.2VVM, open pH automatic control device during fermentation, use HCl or Ca(OH ) 2 Control the pH value of the fermentation brot...
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