The improved purification technique for isolation of the novel CGTase from the alkaliphilc strain Caldalkalibacillus mannanilyticus IB-OR17-B1

Abstract

Cyclodextrin-glucanotransferase (CGTase, EC 2.4.1.19) is a multifunctional enzyme that catalyzes many enzymatic reactions including cyclization, binding, disproportionation and hydrolysis reactions, playing an important role in the enzymatic synthesis of compounds that are widely used in agriculture, pharmaceuticals, food, chemical and biotechnology industries. The present research is aimed to optimize the purification protocol for the extracellular CGTase of alkaliphilc bacterial strain Caldalkalibacillus mannanilyticus IB-OR17-B1 guaranteeing the enzyme homogeneity and its high yield. The improved combination of ultrafiltration and corn-starch (5% w/v) affinity sorption techniques resulted to mild and rapid isolation of electrophoritically homogenic enzyme at 18 × increase of its specific activity and yield 56%. The developed two-step procedure instead the practiced tree-step one using commonly ion-exchange chromatography as final purification technique highly contributes in advance of cost-effectiveness for industrial production and isolation of valuable CGTases.

Keywords:

• Alkaliphilic bacilli
• Caldalkalibacillus mannanilyticus
• CGTase
• cyclodexdrins
• purification

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Disclosure statement

All the authors declare that they have no conflict of interest.

Availability of data material

All data generated or analyzed during this study are included in this published article.

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Author contribution

The concept, main research, data analysis and spelling were performed by P. Milman; E. Gilvanova and G. Aktuganov contributed into special researches and the data preparation. G. Aktuganov is responsible for the editing and process documentation.

Ethical approval

The article does not contain any studies with human participants or animals performed by any of the authors.

Additional information

Funding

The study was supported within framework of the state assignment No. 075-00326-19-00 of the Ministry of Science and Higher Education of the Russian Federation for the topic No. АААА-А18-118022190098-9.