Preparation method of transglucosidase
A technology of transglucosidase and spore suspension, which is applied in the field of preparation of transglucosidase, can solve the problems of low isomalto-oligosaccharide transglycosidase and the like, and achieve the effects of shortening reaction time, improving reaction rate and reducing production time.
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Embodiment 1
[0024] Such as figure 1 As shown, a preparation method of transglucosidase, according to the production steps described in the content of the invention, aspergillus niger is cultivated on a slant for 7 days to prepare mature spores, and sterile water is added to prepare a slant spore suspension. In the PDY medium shake flask, culture at 32℃, 150r / min for 48h, and put the spore suspension in the shake flask in a 500ML shake flask containing 100ML liquid medium at a ratio of 10%. The weight percentages of the components of the optimized liquid fermentation medium are as follows: sucrose 10%, peptone 2%, yeast extract 0.3%, K 2 HPO 4 0.5%, N-methyl-D-glucosamine 0.05%, NaCl0.15%, MgSO 4 0.02%, the rest is water, prepared by mixing uniformly, initial pH 6.05, cultured at 28℃, 29℃, 30℃, 31℃, 32℃, and 180r / min for 60h, and centrifuged 5000r / min for fermentation broth , 2 minutes, take the supernatant to test the enzyme activity, as shown in Table 1. Enzyme activity detection method (...
Embodiment 2
[0029] Such as figure 1 As shown, a preparation method of transglucosidase, according to the production steps described in the content of the invention, aspergillus niger is cultivated on a slope for 7 days to prepare mature spores, and sterile water is added to prepare a suspension of slope spores. In the PDY medium shake flask, culture at 32℃, 150r / min for 48h, and put the spore suspension in the shake flask in a 500ML shake flask containing 100ML liquid medium at a ratio of 10%. The weight percentages of the optimized liquid fermentation medium components are as follows: 12% sucrose, 2% peptone, 0.1% yeast extract, K 2 HPO 4 0.1%, N-methyl-D-glucosamine 0.05%, NaCl0.5%, MgSO 4 0.02%, the rest is water, prepared by mixing uniformly, initial pH 6.02, cultured at 32°C, 180r / min for 60h. Centrifuge the fermentation broth at 5000r / min for 2 minutes, take the supernatant to test the enzyme activity, and use different inducer N-methyl-D-glucosamine dosage (weight percentage) as a co...
Embodiment 3
[0034] Such as figure 1 As shown, a preparation method of transglucosidase, according to the production steps described in the content of the invention, aspergillus niger is cultivated on a slant for 7 days to prepare mature spores, and sterile water is added to prepare a slant spore suspension. In the PDY medium shake flask, culture at 32°C and 150r / min for 48h, and put the spore suspension in the shake flask in a 500ML shake flask containing 100ML liquid medium at a ratio of 10%. The weight percentages of the optimized liquid fermentation medium components are as follows: sucrose 5%, peptone 8%, yeast extract 0.5%, K 2 HPO 4 1%, N-methyl-D-glucosamine 0.05%, NaCl1%, MgSO 4 0.05%, the rest is water, prepared by mixing uniformly, initial pH 6.01, incubate at 32℃, 180r / min for 60h, centrifuge the fermentation broth at 5000r / min for 2 minutes, take the supernatant to test the enzyme activity, prepare The activity of the obtained crude enzyme solution was 20850u / ML.
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