Production and purification of CGTase of alkalophylicBacillus isolated from Brazilian soil |
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Authors: | Matioli Graciette Zanin Gisella M. Gljimarães Manoel F. De moraes Flávio f. |
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Affiliation: | (1) Chemical Engineering Department, State University of Marin’a, Av. Colombo, 5790-Bl. E46-09, 87020-900 Maringá, PR, Brazil;(2) Biochemistry Department, Federal University of Paraná, P. O. Box 19046, 81531-990 Curitiba-PR, Brazil |
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Abstract: | Alkalophylic bacilli that produce cyclodextringlycosyltransferase (CGTase) were isolated from Brazilian soil, with a scheme of two plating steps. In the first step, the bacterial isolate forms a halo in the cultivation medium that contains γ-cyclodextrin (CD) complexing dyes. The CGTase of an isolate was purified 157-fold by biospecific affinity chromatography, with β-CD showing a mol wt of 77,580 Daltons. It produces a γ- to β-CD ratio of 0.156 and a small amount of α-CD, using maltodextrin 10% as substrate, at 50°C, pH 8.0 and 22 h reaction time, reaching 21.4% conversion of the substrate to cyclodextrins. In the second screening step, the isolates chosen give larger halos with β-CD complexing dyes, and smaller halos with β-CD complexing dyes, leading to a 30% improvement in γ-CD selectivity, although at lower total yield for cyclodextrins (11.5%). |
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Keywords: | KeywordHeading" >Index Entries Cyclodextringlycosyltransferase CGTase cyclodextrin alkalophylic bacillus screening |
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