Abstract
In producing recombinant β-glucosidase in Escherichia coli by high-cell density cultivation (HCDC), insufficient soluble oxygen is always a problem. To address it, Vitreoscilla hemoglobin (VHb) was introduced into Escherichia coli by the bicistron and T₇ promoter expression systems, to improve soluble oxygen by bacterial cells and thereby to enhance the biomass and recombinant β-glucosidase production. In the case of bicistron expression system, cell density in shaking flask reached OD₆₀₀=(4.24±0.29), 35.03% higher than that of the control without VHb. Correspondingly, the maximum activity of β-glucosidase co-expressed with VHb was (9.78±0.55) U/mL, 25.38% higher than that of the control. In a 3-L fermentor, the maximum activity of β-glucosidase was 141.23 U/mL, 35.57% higher than that of the control. In contrast, the activity of β-glucosidase co-expressed with VHb under T₇ promoter was lower than that of the control, either in flask or in fermentor. Co-expressing β-glucosidase with VHb using the bicistron expression system may improve the tolerance of E. coli to insufficient soluble oxygen and thus promote the bacterial biomass and the enzyme yield.
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