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

C-niveau - KVUC Bild. EP2316929B1 - Maltogenic alpha-amylase variants - Google Patents. Effect of storage temperature on -amylase activity from .

process. DNA from the CGTase source organism (E. coli Cyclodextrin glucanotransferase (CGTase, EC. 2.1.1.19) produced using new alkaliphile Microbacterium terrae KNR 9 has been purified to homogeneity in a single step by the starch adsorption method. The specific activity of the purified CGTase was 45 U/mg compared to crude 0.9 U/mg.

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2013a, b). The culture supernatant (0.1 mL) was mixed with 0.9 mL of 5% (w/v) soluble starch in 50 mM phosphate buffer (pH 6.0) and incubated at 40 °C for 10 min. D-xylose. The CGTase activity was assayed by the method of Nomoto et al.

CGTase activity was assayed as described by Kato and Horikoshi [ 5 ]. One unit of enzyme activity was defined as the amount of enzyme that forms 1 μmol of γ-CD from soluble starch in 1 min. Alkaline phosphatase was assayed by the method of Yamane et al. [ 9 ].

Cyclodextrin glucanotransferase (CGTase, EC 2.4.1.9) is an unique enzyme capable of converting starch and related substrates into cyclodextrins (CDs). In this paper, we report an one step gel Estimation of CGTase Activity CGTase activity was measured using phenolphthalein β-cyclodextrin complexation method20. Enzyme solution dialyzed against buffer before enzyme assay. When required, different salts such as NaCl, KCl, CaCl2 and NH4Cl were introduced in the reaction mixture in 10-70 mM final The extracellular β-CGTase activity of CGTd4 reached 571.2 U/mL after 57.5 h of fermentation (Fig.

Beneficial Activity by Bacillus subtilis. Ramesh C Ray ⋅ Manas R Swain Häftad Production of CGTase from Bacillus subtilis. Bhargavi Kumaraswamy. 689 

BL-31 was stimulated by presence of Mn 2+ which resulted in increased yield of glycosylated naringin from 80.2% to 92.1% . However, some other metal ions such as Zn 2+ , Cu 2+ and Fe 2+ are inhibitors for most CGTase [ 158 ]. An unit of CGTase activity was defined as the amount of enzyme that produces 1 mmol of b-CD per minute, under the assay conditions. 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.

Maximum CGTase activity reached at 24 h of incubation in all media except medium supplemented with ammonium sulphate (Fig.
Thermodynamics is the study of

CGTase activity. The introduction of an ionic interaction at the Ca-I site via the mutation N132R disrupted CGTase catalytic activity. Conversely, the variant N28R, which has an additional ionic interaction at the Ca-II site, displayed increased cyclization activity. However, thermostability was not affected. The eluted fractions containing CGTase activity were pooled and then brought to 80% ammonium sulfate saturation.

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.
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The activity of GSTs is dependent upon a steady supply of GSH from the synthetic enzymes gamma-glutamylcysteine synthetase and glutathione synthetase. As 

The CGTase activity was increased in the presence of metal ions (5 mM): Ca+2 (130 %), Mg+2 (123 %), Mn+2 (119 %) and Co+2 (116 %). The enzyme activity was strongly inhibited in the presence of Hg+ The β-CGTase from alkalophilic Bacillus sp. N-227 was separately mutagenized to give three site-directed β-CGTase mutants, Y127F, R254F and D355R, that showed enhanced cyclization activity towards a starch substrate from 1.64 to 2.1-folds. Kinetic studies indicate that the mutants had higher affinity towards the substrate than the wild type The optimum temperature of CGTase activity increased from 50 to 60 °C after the stabilizer application.

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, 

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. 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. The introduction of an ionic interaction at the Ca-I site via the mutation N132R disrupted CGTase catalytic activity.

All these activities share the  23 Sep 2016 megaterium in a submerged fermentation. Afterwards, the CGTase was purified partially and its activity to synthesize α-, β-, and γ-CDs was  It was reported that CGTases are active on both starch fractions: amylose and amylopectin but the high amylopectin content were preferred for CGTase activity. (  General CGTase assay (iodine method).The activity of CGTase was assayed using amylose AS-30 as a substrate by measuring the decrease in iodine- staining  CGTase activity and β-cyclodextrin production were determined by phenolphthalein assay method described by Goel and Nene [15] with minor modification. Isolate showing highest CGTase activity was characterized morphologically and biochemically.