Method for preparing internally tangent-beta-1,3-1,4-dextranase from recombination escherichia coli XM-LU
A technology of recombinant Escherichia coli and glucanase, which is applied in the field of bacterial culture medium and fermentation technology, can solve the problems of microbial growth inhibition, limitation of continuous cell growth and protein expression, and few reports of β-glucanase. The effect of reducing costs and increasing product output
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Embodiment 1
[0021] Example 1: Different carbon sources were selected respectively, and the effects of different concentrations of carbon sources on the growth of bacteria and the synthesis of β-glucanase were investigated. Batch culture conditions: ventilation rate 1L / (L·min), stirring rate 400r / min, fermentation temperature 37°C. Other components in the medium: yeast powder 24g L -1 , NaCl 10g·L -1 、KH 2 PO 4 2.4g L-1, K 2 HPO 4 12.5g·L -1 , Kanamycin 0.05g·L -1 , and the rest is water. The results are shown in Table 1. The results showed that when sucrose and glucose were used as carbon sources, the enzyme activities were lower, and further decreased with the increase of the added concentration. The enzyme production effect was the best when wheat flour was used as carbon source, followed by barley flour and glycerol. However, since most of the ingredients in wheat flour are insoluble in water, it is difficult to separate the cells from it after fermentation, which will have a...
Embodiment 2
[0024] Embodiment 2: with glycerol (10g L -1 ) and wheat flour (50g·L -1 ) is the carbon source, and under the intermittent culture conditions described in Example 1, the effects of different nitrogen sources on cell growth and β-glucanase synthesis were investigated. Other components in the medium except carbon and nitrogen sources: NaCl 10g L -1 、KH 2 PO 4 2.4g·L -1 、K 2 HPO 4 12.5g·L -1 , Kanamycin 0.05g·L -1 , and the rest is water. The results are shown in Table 2.
[0025] Table 2 Effect of nitrogen source on bacterial growth and enzyme production
[0026]
[0027] The results showed that when inorganic nitrogen was used as the only nitrogen source, the biomass was very low, which limited the synthesis of enzymes; however, when yeast powder and corn steep liquor were used as organic nitrogen sources and glycerol, the enzyme activity was significantly improved.
Embodiment 3
[0028] Embodiment 3: with glycerol (10g L -1 ) is the carbon source, and the intermittent culture conditions as described in Example 1 were used to investigate the effect of the combination of organic and inorganic nitrogen sources on cell growth and β-glucanase synthesis. The other components in the culture medium are as in Example 2 except carbon and nitrogen sources. The results are shown in Table 3.
[0029] Table 3 Effect of combination of organic and inorganic nitrogen on bacterial growth and enzyme production
[0030]
[0031] It can be seen from Table 3 that the combination of yeast powder and corn steep liquor and inorganic nitrogen sources can promote the synthesis of enzymes, especially yeast powder and NaNO 3 (1.20∶0.72) when compounded, the enzyme activity reached nearly 300U / mL, corn steep liquor and NaNO 3 (2.4:1.2) when compounded, the enzyme activity reached 306.51U / mL. However, the stimulating effect of corn steep liquor on the growth of bacteria is mo...
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