Method for efficiently introducing L-arabinose isomerase
An arabinose and isomerase technology, applied in the field of efficient induction of L-arabinose isomerase, can solve the problems of unfavorable L-arabinose isomerase activity expression, fast protein expression rate, and difficult to fold inclusion bodies, etc. The effect of reducing fermentation production costs, reducing production costs, and promoting folding
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Embodiment 1
[0018] Example 1 Adding D-galactose to promote the high expression of L-arabinose isomerase of A. cellulolyticus
[0019] The construction method of Escherichia coli BL21(DE3) containing A. cellulolyticus L-arabinose isomerase is as reported in the literature (Appl Microbiol Biotechnol, 2010, 86(4): 1089-1097), and the host cell used is large intestine Bacillus BL21(DE3), the source of the exogenous L-arabinose isomerase gene is Acidothermus cellulolytics ATCC 43068, and the expression vector plasmid is pET-22b(+) of the lactose operon.
[0020] When the recombinant bacteria were cultured in LB medium at 37°C until the light absorption value at 600nm was 0.6, 1mmol / L conventional inducer IPTG was added, and enzyme substrate D-galactose of different concentrations was added at the same time, induced at 30°C for 4h, and the bacteria were collected. The body was tested for fermentation enzyme activity, and the experimental results are shown in Table 1.
[0021] Table 1 Adding D-...
Embodiment 2
[0025] Example 2 Adding D-galactose to promote the high expression of L-arabinose isomerase of Bacillus stearothermophilus
[0026] The construction method of Escherichia coli BL21(DE3) containing Bacillus stearothermophilus L-arabinose isomerase is as reported in the literature (J Sci Food Agric 2010, 90(8): 1327-1333), and the host cell used is large intestine Bacillus BL21(DE3), the source of the exogenous L-arabinose isomerase gene is Bacillus stearothermophilus IAM 11001, and the expression vector plasmid is pET-22b(+) of the lactose operon.
[0027] When the recombinant bacteria were cultured in LB medium at 37°C until the light absorption value at 600nm was 0.6, 1mmol / L conventional inducer IPTG was added, and enzyme substrate D-galactose of different concentrations was added at the same time, induced at 30°C for 4h, and the bacteria were collected. The body was tested for fermentation enzyme activity, and the experimental results are shown in Table 2.
[0028] Table 2...
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