Effect of PEG and other additives on cyclodextrin production by Bacillus macerans cyclomaltodextrin-glycosyl-transferase

Delbourg, M. F.; Drouet, Ph.; Moraes, Fabiane de; Thomas, Daniel; Barbotin, Jean‐Noël

doi:10.1007/bf00133016

Cited by 10 publications

(10 citation statements)

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“…Finally, we checked the stability of different CGTase preparations in the presence of ethanol, a cosolvent proposed by some authors for the improvement of the CGTase in the production of cyclodextrins (Delbourg et al, 1993; Shiraishi et al, 1989; Abraham, 1995; Mori et al, 1995). Figure 4 shows that the glyoxyl‐agarose immobilized enzyme was much more stable than any other preparation under these conditions (by more than a 100‐fold factor).…”

Section: Resultsmentioning

confidence: 99%

“…Different organic compounds have been used in many instances to improve the CD yields or to alter the percentage of the CD produced. Several authors have intended to explain this effect (Delbourg et al, 1993; Shiraishi et al, 1989; Abraham, 1995; Mori et al, 1995). Taking advantage of the high stability of the CGTase‐glyoxyl‐agarose preparation in the presence of ethanol even at a concentration of 30%, it was possible to compare the effect of the ethanol in the reaction rate of production of β‐CD from starch or maltodextrins with the β‐CD degradation with maltose as acceptor.…”

Section: Resultsmentioning

confidence: 99%

“…The resistance to the deleterious effect of ethanol on enzyme stability has allowed the study of the effect of ethanol in the production and degradation activities of CGTase, showing that ethanol strongly inhibits the degradation reaction while having a much more reduced effect on the synthetic activity. Thus, although other effects of cosolvents may be not discarded (Delbourg et al, 1993; Shiraishi et al, 1989; Abraham, 1995; Mori et al, 1995), the inhibition of the hydrolysis of CD by ethanol has been clearly established.…”

Section: Discussionmentioning

confidence: 99%

“…The reaction conditions could be modified to improve the yield and selectivity at industrial scale (Blackwood and Bucke, 2000; Gawande and Patkar, 2001; Plou et al, 2002). For example, ethanol and other cosolvents have been shown to permit an increment in the CD production, although the reasons for this increment are still under debate (Delbourg et al, 1993; Shiraishi et al, 1989; Abraham, 1995; Mori et al, 1995). However, this kind of reaction medium may have strong deleterious effects on enzyme stability.…”

Section: Introductionmentioning

confidence: 99%

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Immobilization and Stabilization of a Cyclodextrin Glycosyltransferase by Covalent Attachment on Highly Activated Glyoxyl-Agarose Supports

et al. 2006

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Covalent immobilization of cyclodextrin glycosyltransferase on glyoxyl-agarose beads promotes a very high stabilization of the enzyme against any distorting agent (temperature, pH, organic solvents). For example, the optimized immobilized preparation preserves 90% of initial activity when incubated for 22 h in 30% ethanol at pH 7 and 40 degrees C. Other immobilized preparations (obtained via other immobilization protocols) exhibit less than 10% of activity after incubation under similar conditions. Optimized glyoxyl-agarose immobilized preparation expressed a high percentage of catalytic activity (70%). Immobilization using any technique prevents enzyme inactivation by air bubbles during strong stirring of the enzyme. Stabilization of the enzyme immobilized on glyoxyl-agarose is higher when using the highest activation degree (75 micromol of glyoxyl per milliliter of support) as well as when performing long enzyme-support incubation times (4 h) at room temperature. Multipoint covalent immobilization seems to be responsible for this very high stabilization associated to the immobilization process on highly activated glyoxyl-agarose. The stabilization of the enzyme against the inactivation by ethanol seems to be interesting to improve cyclodextrin production: ethanol strongly inhibits the enzymatic degradation of cyclodextrin while hardly affecting the cyclodextrin production rate of the immobilized-stabilized preparation.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Immobilization and Stabilization of a Cyclodextrin Glycosyltransferase by Covalent Attachment on Highly Activated Glyoxyl-Agarose Supports

et al. 2006

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“…Previous studies revealed that the addition of PEG leads to an increase in the thermal stability of some enzymes (31)(32)(33). Therefore, the effect of PEG 2000 on the thermal sensitivity of Fuc-TVII was examined by preincubating at different temperatures for 1 h with or without PEG 2000, followed by measurement of its residual activity at 37°C.…”

Section: Preparation Of Fuc-tvii-mentioning

confidence: 99%

Enzymatic Characterization of Human α1,3-Fucosyltransferase Fuc-TVII Synthesized in a B Cell Lymphoma Cell Line

Shinoda

Morishita

Sasaki

et al. 1997

Journal of Biological Chemistry

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The human ␣1,3-fucosyltransferase, Fuc-TVII, a key enzyme in the biosynthesis of selectin ligands, was expressed as a soluble protein-A chimeric form in a human B cell lymphoma cell line, Namalwa KJM-1, and purified using IgG-Sepharose. The enzymatic properties of recombinant soluble Fuc-TVII were then examined. Its enzyme activity was highest at pH 7.5, and the presence of 25 mM Mn 2؉ was required for full activity. Fuc-TVII exhibits an acceptor specificity restricted to ␣2,3-sialylated type 2 oligosaccharides, and the apparent K m values for ␣2,3-sialyl lacto-N-neotetraose and GDP-fucose were 3.08 mM and 16.4 M, respectively. The inhibitory effects of various nucleotides on the activity of Fuc-TVII reflected its donor specificity for the nucleotide portion of GDP. Fuc-TVII was demonstrated to be useful for the synthesis of a sialyl Lewis x hexasaccharide from lacto-N-neotetraose in combination with an ␣2,3-sialyltransferase, ST3Gal IV. Polyethylene glycols enhanced the thermal stability of Fuc-TVII, leading to increased formation of the reaction product.Cell surface glycoproteins and glycolipids containing ␣1,3-fucosylated lactosaminoglycans play important roles in cell-cell interaction and cell migration in connection with physiological and pathological processes such as embryogenesis, cancer metastasis, lymphocyte trafficking, and immune responses (1, 2). In particular, the sialyl Lewis x (sLe x ; NeuAc␣2-3Gal␤1-4(Fuc␣1-3)GlcNAc) oligosaccharide structure and related structures have evoked considerable interest because they were demonstrated to serve as ligands for the three known selectins (E-, P-, and L-selectins), which are cell adhesion molecules involved in the recruitment of leukocytes to lymphoid tissues and the sites of inflammation (2-6).The biosynthesis of ␣1,3-fucosylated lactosaminoglycans requires the actions of several glycosyltransferases. Among them, an ␣1,3-fucosyltransferase (Fuc-T) is considered to be the key enzyme, which catalyzes the transfer of fucose from GDPfucose to N-acetylglucosamine via an ␣1,3-linkage. The expression-cloning approach led to the cloning of the first human ␣1,3-Fuc-T, Fuc-TIII (7). Subsequently, low stringency hybridization-cloning and expression-cloning approaches revealed the cDNAs for three additional members, Fuc-TIV, Fuc-TV, and Fuc-TVI (8 -13). Detailed analyses have shown that Fuc-TIII, Fuc-TIV, and Fuc-TVI correspond to Lewis-type, myeloid-type, and plasma-type enzymes, respectively (7-10, 14). Biochemical analyses involving enzyme preparations extracted from human tissues and/or the respective recombinant proteins revealed their acceptor specificities, pH optima, kinetic properties, cation requirements, and N-ethylmaleimide (NEM) 1 sensitivities (15-20).We (21) and Natsuka et al. (22) previously cloned a novel human ␣1,3-Fuc-T, Fuc-TVII, expressed in myeloid lineage cells that had never been characterized biochemically. We presented evidence of its involvement in the biosynthesis of an E-selectin ligand. Recent studies have also demonstrated its involveme...

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Complex Formation of Unsaturated Cyclodextrin Solutions with Various Polymers

Kildemark

Larsen

Zimmermann

1996

Proceedings of the Eighth International Symposium on Cyclodextrins

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Effect of PEG and other additives on cyclodextrin production by Bacillus macerans cyclomaltodextrin-glycosyl-transferase

Cited by 10 publications

References 18 publications

Immobilization and Stabilization of a Cyclodextrin Glycosyltransferase by Covalent Attachment on Highly Activated Glyoxyl-Agarose Supports

Immobilization and Stabilization of a Cyclodextrin Glycosyltransferase by Covalent Attachment on Highly Activated Glyoxyl-Agarose Supports

Enzymatic Characterization of Human α1,3-Fucosyltransferase Fuc-TVII Synthesized in a B Cell Lymphoma Cell Line

Complex Formation of Unsaturated Cyclodextrin Solutions with Various Polymers

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