A CGTase with high coupling activity using γ-cyclodextrin isolated from a novel strain clustering under the genus Carboxydocella. Gulshan Kazi, Zubaida LU; Lundemo, Pontus LU; Fridjonsson, Olafur H; Hreggvidson, Gudmundur O; Adlercreutz, Patrick LU and Nordberg Karlsson, Eva LU () In Glycobiology 25 (5). p.514-523

γ-Cyclodextrin glycosyltransferase (γ-CGTase) catalyzes the biotransformation of low-cost starch into valuable γ-cyclodextrin (γ-CD), which is widely applied in biotechnology, food, and pharmaceutical industries.

Results: Using the enzyme β-cyclodextrin glycosyltransferase (β-CGTase) as a reporter protein and B. subtilis CCTCC M 2016536 as the host, nine plasmids equipped with single promoters were screened using shake-flask cultivation. The plasmid containing the P amyQ' promoter produced the greatest extracellular β-CGTase activity; 24.1 U/mL. soluble γ-CGTase activity had reached 5.51 U·mL−1.In addition, the ratio of extracellular γ-CGTase activity to total γ-CGTase activity decreased from 71.7 to 55.0% when the concentration of β-cyclodextrin increased from 0 to 10 mM. The effect of glycine concentration on γ-CGTase production by E. coli Figure 2 Time courses of the biomass concentration of Bacillus sp. G1 in continuous culture at various dilution rates. After batch culture for 12h, continuous culture was operated at 35oC with aeration of 1v/v/m and agitation at 150rpm for 72h. - "Impact of dilution rate on CGTase activity and productivity from an alkalophilic Bacillus sp.

Cgtase activity

The CGTase activity profile was successfully maintained atsteady state using dilution rates of 0.03 to 0.3h-1. Maximum CGTase activity was obtained atlow dilution rate range (0.03-0.07h-1). CGTase activity decreased from 26.4 to 12.4 U/mlwhen the dilution rate was increased to 0.3h-1… CONTINUE READING Enzyme activity of CGTase from Alkalophilic Bacillus sp. BL-31 was stimulated by presence of Mn 2+ which resulted in increased yield of glycosylated naringin from 80.2% to 92.1% .

nitrogen source for CGTase production. Ca2+ influences the CGTase production and Zn2+ inhibits the enzyme. High CGTase activity was observed at 24 h of.

The optimisation of this growth medium was carried out using response surface methodology. CGTase was expressed by recombinant K. phaffii through pH maintenance in range of 5.5–7.0. β‐CGTase activity increased to 122.0 U/mL after optimization of glycerol, phosphate buffer, pH value, ammonium sulfate, temperature, methanol, and additives based on BSM, establishing a modified defined medium. 23 Aug 2017 When expression was induced at 25 °C for 32 h, and then the temperature was shifted to 30 °C, the extracellular α-CGTase activity at 90 h was 45 Catalytic activities[edit].

CGTase was expressed by recombinant K. phaffii through pH maintenance in range of 5.5–7.0. β‐CGTase activity increased to 122.0 U/mL after optimization of glycerol, phosphate buffer, pH value, ammonium sulfate, temperature, methanol, and additives based on BSM, establishing a modified defined medium.

Gulshan Kazi, Zubaida LU; Lundemo, Pontus LU; Fridjonsson, Olafur H; Hreggvidson, Gudmundur O; Adlercreutz, Patrick LU and Nordberg Karlsson, Eva LU () In Glycobiology 25 (5). p.514-523 Activity and stability characteristics of an alkaline active cyclodextrin glycosyltransferase (CGTase) enzyme from the alkaliphilic Bacillus agaradhaerens LS-3C strain are reported. The enzyme displays unusually high amylolytic activity in relation to the cyclization activity. The standard CGTase activity assays described above was used to determine the residual activity of each enzyme (Jeang et al. 2005). Kinetic assays The K m and V max values for the purified enzymes were determined by incubating 100 µL of enzyme (0.5 µg) in 200 µL of 0.2 M phosphate buffer (pH 6.0) with soluble starch solution (0.4–6.0 mg/mL) at 60 °C for 10 min. CGTase activity was assayed as described by Kato and Horikoshi .

The specific activity was expressed in units of activity by milligram of protein. Protein concentration was estimated according to Hartree (12), using bovine serum albumin as pattern. 2017-10-10 · The β-CGTase activity in the transformant reached 3885.1UmL-1, which is the highest value reported so far, at 28°C, 6% inoculum ratio, and 1.5% methanol addition following 24h of incubation. The recombinant CGTase showed high specific activity at 80°C without any γ-CGTase activity, and had good stability in a wide pH and temperature range. CGTase which showed high specific activity at 80°C without any γ-CGTase activity (Zhang et al. 2017).
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As 24 Apr 2017 Cyclodextrin glycosyltransferase (CGTase) catalyzes the formation of The synergistic effect of individual parameters of CGTase activity and 10 May 2013 One unit of CGTase activity was defined as the amount of enzyme that produces 1 µmol of β-CD.min-1. For partial purification, the enzyme was 15 Nov 2013 stability compared with free enzyme. The optimum pH for enzyme activity was pH 8 and pH 7.5 for free and immobilized CGTase, respectively, A CGTase with high coupling activity using γ-cyclodextrin isolated from a novel strain clustering under the genus Carboxydocella.
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At this point the total soluble γ-CGTase activity had reached 5.51 U·mL − 1. In addition, the ratio of extracellular γ-CGTase activity to total γ-CGTase activity decreased from 71.7 to 55.0% when the concentration of β-cyclodextrin increased from 0 to 10 mM.

G1 in continuous culture" Some of the enzyme activity was also observed in the periplasmic and the intracellular fractions. When the mature CGTase G1 gene (without signal peptide) was cloned into pQE and pWH expression vectors and transformed into E. coli, almost all of the CGTase activity was detected intracellularly (data not shown). 2016-11-01 · glycosyltransferase (CGTase, EC 2.4.1.19, CAS 9030-09-5) preparation, residual starch, linear maltooligosaccharides, a-CD does not contain any CGTase activity because the enzyme is . CGTase activity.