Method for immobilizing lipase B from candida antarctica fermented supernatant fluid
A technology of Candida Antarctica, fermentation supernatant, applied in microorganism-based methods, immobilized on/in organic carriers, biochemical equipment and methods, etc., can solve difficult large-scale production, CALB enzyme activity recovery Reduce the rate of reduction, low catalytic activity and other problems, to achieve the effect of weakening the exclusion effect, easy to judge the end point, and enhancing the catalytic ability
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Embodiment 1
[0035] The activity of lipase B in the fermentation supernatant of Candida antarctica is 2256U / mL, the volume is 500L, and the temperature is pre-cooled to 20°C in a jacketed stirring tank.
[0036] The diameter of the pre-column is 30cm, and the height is 200cm (diameter-to-height ratio 1:6.67). Take 100L of NKA-II macroporous resin as the pre-column filler (the volume ratio of resin to fermentation supernatant is 1:5), add 50L methanol to soak, and use 50L Carry out wet packing of methanol (column packing volume 0.71BV), use 200L methanol as the mobile phase for cleaning, flow rate 150L / h (flow rate 1.06BV / h), use 300L distilled water to wash out methanol after methanol cleaning, flow rate 150L / h (flow rate 1.06BV / h).
[0037] The diameter of the adsorption column is 20cm, and the height is 200cm (the ratio of diameter to height is 1:10). Take 50L of HPD600 macroporous resin as the filler of the adsorption column (the volume ratio of resin to fermentation supernatant is 1:10...
Embodiment 2
[0052] The activity of lipase B in the fermentation supernatant of Candida antarctica is 2328U / mL, the volume is 5000L, and the temperature is pre-cooled to 15°C in a jacketed stirring tank.
[0053] The pre-columns were prepared and pretreated according to the specifications and methods in Example 1, and a total of 6 pre-columns were prepared and connected in parallel.
[0054] The diameter of the adsorption column is 30cm, and the height is 200cm (diameter-height ratio 1:6.67). A total of 6 columns are prepared, which are respectively connected in series to the pre-column, and 105L of HPD600 macroporous resin is respectively taken as the adsorption column filler (the total volume ratio of the resin to the fermentation supernatant is 1:8), add 50L methanol for soaking, use 50L methanol for wet packing (column packing volume 0.74BV), use 200L methanol as mobile phase for cleaning respectively, flow rate 150L / h (flow rate 1.06BV / h), methanol cleaning Finally, 300L of distilled ...
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