Middle and high temperature resisting exopoly water insoluble polysaccharide protoplast fusion engineering bacterium and application thereof
A technology of protoplast fusion and water-insoluble polysaccharides, which is applied in the direction of bacteria, microorganisms, and drilling compositions, can solve the problems of polysaccharides easily split into fragments and poor toughness, and achieve the effect of improving oil recovery
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Embodiment 1
[0042] Embodiment 1, the construction of protoplast fusion engineering bacteria
[0043] Step 1: Screening of Parental Strains
[0044] The screening method of the parent strain JD is as follows: according to the conventional strain screening method, the water sample collected from Jilin Oilfield was diluted 10,000 times, spread on the agar plate containing 4% (v / v) molasses, and cultured at 30°C for 24 hours. Connect the isolated single colonies of various microorganisms into 50mL 4% molasses liquid medium, and place them for anaerobic culture at 30°C for 1 day to judge the production status of insoluble polymers. After multiple separations and purifications, a water-producing strain was obtained. JD strains of insoluble polymers. The strain can utilize monosaccharides to synthesize extracellular water-insoluble polysaccharides at a temperature lower than 37°C.
[0045] The screening method of the parent strain AJ is as follows: According to the conventional strain screenin...
Embodiment 2
[0056] Example 2, the influence of temperature on the extracellular water-insoluble polysaccharide produced by protoplast fusion engineering bacteria ZR3
[0057]Inoculate JD bacteria and protoplast fusion engineering bacteria ZR3 into 50mL liquid complete medium (1% peptone, 1% glucose, 0.5% yeast powder, 0.5% beef extract, 0.5% NaCl, pH value 7.2), the inoculation amount is 1%, respectively placed at 20°C, 30°C, 35°C, 40°C, 42°C, and 45°C for static culture for 2 days.
[0058] After the cultivation, extracellular water-insoluble polysaccharides were obtained by filtering with filter paper, washed with distilled water and weighed. The effect of temperature on the production of extracellular water-insoluble polysaccharides by protoplast fusion engineered bacteria ZR3 is shown in figure 1 , the maximum temperature of extracellular water-insoluble polysaccharide produced by protoplast fusion engineering strain ZR3 was 45°C, and the production of extracellular water-insoluble p...
Embodiment 3
[0059] Example 3, Effect of Salt Concentration on the Production of Extracellular Water-Insoluble Polysaccharides by Protoplast Fusion Engineering Bacteria ZR3
[0060] Take 30.5 mL of activated fusion engineered bacteria ZR and inoculate them in 50 mL of liquid complete medium containing 1000 mg / L, 2000 mg / L, 3000 mg / L, 5000 mg / L, 10000 mg / L, 20000 mg / L and 50000 mg / L NaCl ( 1% peptone, 1% glucose, 0.5% yeast powder, 0.5% beef extract, pH 7.2), cultured at 40°C for 2 days
[0061] After the cultivation, extracellular water-insoluble polysaccharides were obtained by filtering with filter paper, washed with distilled water and weighed. The effect of salt concentration on the production of extracellular water-insoluble polysaccharides by protoplast fusion engineered bacteria ZR3 is shown in figure 2 , the protoplast fusion engineering strain ZR3 can still produce polysaccharides at a high salt concentration of 50000mg / L, and the sugar production is relatively high, and the pro...
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