Production, purification, and properties of dextransucrase from Leuconostoc mesenteroides NRRL B-512F

John F. Robyt, Timothy F. Walseth

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105 Scopus citations

Abstract

The production of dextransucrase fromLeuconostoc mesenteroides NRRL B-512F was stimulated 2-fold by the addition of 0.005% of calcium chloride to the medium; levansucrase levels were unaffected. Dextransucrase was purified by concentration and dialysis of the culture supernatant with a Bio-Fiber 80 miniplant, and by treatment with dextranase followed by chromatography on Bio-Gel A-5m. A 240-fold purification, with a specific activity of 53 U/mg, was obtained. Contaminating enzyme activities of levansucrase, invertase, dextranase, glucosidase, and sucrose phosphorylase were decreased to non-detectable levels. Poly(acrylamide)-gel electrophoresis of the purified enzyme showed only two protein bands, both of which had dextransucrase activity. These bands also gave a carbohydrate stain, indicating that the dextransucrase could be a glycoprotein. Acid hydrolysis, followed by paper chromatography, of the purified enzyme showed that the major carbohydrate was mannose. ConcanavaIin A completely removed dextransucrase activity from solution, confirming the mannoglycoprotein character of the enzyme. Dextransucrase activity was not altered by the addition of 0.008-4 mg/ml of dextran, but its storage stability was increased by the addition of 4 mg/ml of dextran. As previously shown by others, the activity of dextransucrase was decreased by EDTA, and was restored by the addition of calcium ions. Zinc, cadmium, lead, mercury, and copper ions were inhibitory to various degrees.

Original languageEnglish (US)
Pages (from-to)95-111
Number of pages17
JournalCarbohydrate Research
Volume68
Issue number1
DOIs
StatePublished - Jan 1979
Externally publishedYes

Bibliographical note

Funding Information:
*This work was supported by Grant No. DE-03578 from the National Institute of Dental Research, National Institutes of Health, U.S. Public Health Service, and was taken in part from a thesis by T.F.W. submitted to Iowa State University, Ames, Iowa, in partial fulfihnent of the requirements for the Ph.D. degree. tPresent address: Department of PhysioIogy, Vanderbilt University, Nashville, Tennessee 37202, U S.A.

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