Abstract
BackgroundMaltoheptaose as malto-oligosaccharides with specific degree of polymerization, has wide applications in food, medicine and cosmetics industries. Currently, cyclodextrinase have been applied as prepared enzyme to prepare maltoheptaose. However, the yield and proportion of maltoheptaose was lower, which is due to limited substrate and product specificity of cyclodextrinase (CDase). To achieve higher maltoheptaose yield, cyclodextrinase with high substrate and product specificity should be obtained.ResultsIn this study, cyclodextrinase derived from Thermococcus sp B1001 (TsCDase) was successfully expressed and characterized in Bacillus subtilis for the first time. The specific activity of TsCDase was 637.95 U/mg under optimal conditions of 90 °C and pH 5.5, which exhibited high substrate specificity for cyclodextrins (CDs). When the concentration of β-CD was 8%, the yield of maltoheptaose achieved by TsCDase was 82.33% across all reaction products, which exceeded the yields of maltoheptaose in other recent reports. Among malto-oligosaccharides generated as reaction products, maltoheptaose was present in the highest proportion, about 94.55%.ConclusionsThis study provides high substrate and product specificity of TsCDase. TsCDase is able to prepare higher yield of maltoheptaose through conversion of β-CD in the food industry.
Highlights
Maltoheptaose as malto-oligosaccharides with specific degree of polymerization, has wide applications in food, medicine and cosmetics industries
The pH effect on Thermococcus sp B1001 CDase (TsCDase) stability was obtained by incubation at 4 °C and pH 5.0–9.0 for 24 h
Relative activity of TsCDase was stable at pH 7.5–9.0, retaining over 60.0% of maximum activity, suggesting TsCDase was suitable for storage in weak alkaline buffer
Summary
Maltoheptaose as malto-oligosaccharides with specific degree of polymerization, has wide applications in food, medicine and cosmetics industries. To achieve higher maltoheptaose yield, cyclodextrinase with high substrate and product specificity should be obtained. Malto-oligosaccharides are composed of 2–10 glucose units connected by α-1,4 linkages They are considered to be important functional oligosaccharides that are beneficial for digestion and absorption in humans. Due to their good adaptability, malto-oligosaccharides have been widely applied as food additives in the food industry to improve the properties of food products, such as sweetness, hygroscopicity, stability, viscosity, and gelation [1, 2]. Compared with that of maltotriose and maltotetraose, maltoheptaose has a lower osmotic pressure, higher viscosity, better moisturizing effect, and stronger film-forming performance, so that it has been widely applied in the food, medicine, cosmetics, and. An investigation of enzymes with the substrate and product specificity is essential to achieve the mass production of high purity maltoheptaose
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