Reaction method suitable for being catalyzed by solid catalyst
A solid-phase catalyst and reaction technology, applied in the field of reaction, can solve the problems of limited application range, fragmentation and wear, shortened service life, etc., and achieve the effect of wide application range, avoiding fragmentation and wear, and avoiding the decline of vitality
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example 1
[0060] Example 1. Suspension agitation damages the immobilized enzyme carrier
[0061] use Figure 4 In the shown reactor, the currently widely used granular immobilized enzyme resin carrier Sepabeads EC-EP (Grazu V, Abian O, Mateo C, Batista-Viera F, Fernandez-Lafuente R, Guisan JM.Biotechnol Bioeng.2005, 90:597-605; Grazu V, Abian O, Mateo C, Batista-Viera F, Fernandez-Lafuente R, Guisan JM.Biomacromolecules.2003, 4:1495-1501) and granular immobilized glucose isomerase, respectively. Disc stirring (that is, as mentioned above, the granular immobilized enzyme resin carrier and the immobilized glucose isomerase are respectively loaded into the container 53, and the container 53 follows the rotating shaft 21 for centrifugal rotation to obtain the stirring effect) and suspension stirring test, compared The degree of damage to the resin carrier. Sepabeads EC-EP was purchased from Italy Resindion S.R.L company. Granular immobilized glucose isomerase was purchased from Sigma, St...
example 2
[0068] Example 2. The whole piece of open-pore immobilized cells is used as a stirring paddle for continuous stirring reaction to prepare high fructose syrup [1]
[0069] Glucose isomerase catalyzes the reaction that converts glucose into fructose. The preparation of the vector pRSET-lac-MGI4-35-kan expressing glucose isomerase is as follows. According to the sequence design of pGEMT-Easy (Promega), PCR primers are specifically: upstream primer RBS-NdeI: 5'-CATATGTATATCTCCTTCTTGTGTGAAATTG-3'; downstream primer RBS-AlwNI:
[0070] 5'-CAGTGGCTGCTGCCAGTGGCGATAAGTC-3'. Using pGEMT-Easy (Promega) as a template, PCR was performed with the above primers, and a 755bp product was amplified. The PCR conditions were: 50ng pGEMT-Easy (Promega), 0.4 μM RBS-NdeI, 0.4 μM RBS-AlwNI, 50 μM dATP, 50 μM dTTP, 50 μM dCTP, 50 μM dGTP, 20 mM Tris-HCl (pH 8.8), 10 mM KCl, 10 mM ( NH 4 ) 2 SO 4 , 2mM MgSO 4 , 0.1% Triton X-100, 2.5U Pfu DNA polymerase (Promega), the reaction volume was adjuste...
example 3
[0077] Example 3. The whole piece of open-pore immobilized cells is used as a stirring paddle for continuous stirring reaction in series to prepare high fructose syrup
[0078] The preparation of the glucose isomerase-immobilized cell sheet, the determination of its activity, its installation on the reaction tank, the operation of the isomerization reaction and the reaction tank, etc. basically refer to Example 2. In this example, 50 grams of glucose isomerase-immobilized cell sheets were prepared, and the enzyme activity was 5,298 units / gram. Using 2 such as Figure 4 In the shown reaction tanks, reaction tank I and reaction tank II, 23.5 grams of immobilized cell sheet belts with a specific activity of 5,298 units / g were filled in the grid tray 20 respectively, and reacted separately, and the operation was the same as that of Example 2. When the flow rate of the reaction tank I was recorded at 7.1 g / min, the optical rotation (22°C) was -8.68 (equivalent to 48.68% fructose c...
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