Enterobacteria strain FY-07 and method thereof for producing bacterial cellulose by static liquid submerged fermentation
A FY-07, bacterial cellulose technology, applied in the field of biotechnology and biomaterials, can solve the problem of inability to achieve the effect of static culture, and achieve the effect of improving product quality and reducing production costs
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Embodiment 1
[0032] Screening and breeding of Enterobacter sp. FY-07 strain provided by the invention.
[0033] Take 10mL of oilfield production fluid from Jilin Oilfield in 90mL of glucose-containing inorganic salt medium (medium composition: KH 2 PO 4 3.48g / L, Na 2 HPO 4 12H 2 O 1.5g / L, (NH 4 ) 2 SO 4 2g / L, MgSO 4 ·7H 2 O 0.5g / L, yeast extract 0.05g / L, glucose 10g / L, distilled water 1000mL, pH 7.2, sterilized at 105°C for 30min), placed on a 200r / min low-temperature shaker at 30°C for enrichment culture for 5 days. Due to the fine network structure of bacterial cellulose, its colony is not in close contact with the surface of the solid medium, and it is easy to slide on the surface of the solid medium when picking the colony. Therefore, after the above-mentioned enriched culture is streaked on a solid plate, colonies that are not in close contact with the surface of the medium are selected for re-screening; during re-screening, the selected colonies are inoculated into fresh 100...
Embodiment 2
[0035] Morphological and physiological and biochemical characteristics of Enterobacter sp.FY-07 strain.
[0036] Refer to the experimental method of "Bergey's Manual of Systematic Bacteriology" (Vol.Ⅷ) to detect its Gram staining, cell size and shape, presence or absence of spores, growth temperature, growth pH range, and NaCl tolerance. Catalase, M.R. experiment, V.P. experiment, indole production, nitrate reduction, starch hydrolysis, citrate utilization, glucose fermentation and other experiments.
[0037] Enterobacter FY-07 can grow white, round colonies when cultured on LB agar (peptone 10g / L, yeast powder 5g / L, NaCl 5g / L, agar powder 20g / L) plates at 30°C for 24 hours, and the colonies are Granular, with regular edges, 1-2 mm in size and diameter, not in close contact with the culture medium; Gram staining is negative, and the bacteria are short rod-shaped, with a size of 0.5-1.0 μm (width)×1.0-2.5 μm (length), no Spore formation; growth temperature 15~40℃, growth pH ra...
Embodiment 3
[0039] PCR amplification and sequence determination of 16S rRNA gene of Enterobacter sp.FY-07 strain.
[0040] The 16S rRNA gene sequence characteristics of the Enterobacter sp.FY-07 strain provided by the present invention: the FY-07 strain was inoculated in LB medium (peptone 10g / L, yeast powder 5g / L, NaCl 5g / L L), cultivated on a shaker at 30°C (180rpm) for 24 hours, collected the cells by centrifugation, resuspended, added lysozyme and SDS to break the wall, extracted genomic DNA by the phenol-chloroform method, and used the upstream primer (5'-GAGAGTTTGATCCTGGCTCAG-3' ) and downstream primers (5'-AAGGAGGTGATCCA GCCGCA-3'), use this pair of primers to perform PCR amplification of its 16S rDNA gene, and send the amplified primers to Dalian Bao Biological Company for sequencing. The sequence of its 16S rDNA is shown in SEQ ID No .1 shown. The PCR conditions are: 94°C, 5min; 94°C, 45s, 56.2°C, 45s, 72°C, 90s, 30 cycles; 72°C, 9min, 4°C storage. The length of the 16S rDNA ge...
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