Skip to Content
Merck
  • Cloning and characterization of a Weissella confusa dextransucrase and its application in high fibre baking.

Cloning and characterization of a Weissella confusa dextransucrase and its application in high fibre baking.

PloS one (2015-01-21)
Ilkka Kajala, Qiao Shi, Antti Nyyssölä, Ndegwa Henry Maina, Yaxi Hou, Kati Katina, Maija Tenkanen, Riikka Juvonen
ABSTRACT

Wheat bran offers health benefits as a baking ingredient, but is detrimental to bread textural quality. Dextran production by microbial fermentation improves sourdough bread volume and freshness, but extensive acid production during fermentation may negate this effect. Enzymatic production of dextran in wheat bran was tested to determine if dextran-containing bran could be used in baking without disrupting bread texture. The Weissella confusa VTT E-90392 dextransucrase gene was sequenced and His-tagged dextransucrase Wc392-rDSR was produced in Lactococcus lactis. Purified enzyme was characterized using (14)C-sucrose radioisotope and reducing value-based assays, the former yielding K(m) and V(max) values of 14.7 mM and 8.2 μmol/(mg ∙ min), respectively, at the pH optimum of 5.4. The structure and size of in vitro dextran product was similar to dextran produced in vivo. Dextran (8.1% dry weight) was produced in wheat bran in 6 h using Wc392-rDSR. Bran with and without dextran was used in wheat baking at 20% supplementation level. Dextran presence improved bread softness and neutralized bran-induced volume loss, clearly demonstrating the potential of using dextransucrases in bran bioprocessing for use in baking.

MATERIALS
Product Number
Brand
Product Description

Supelco
Methanol, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Sodium chloride solution, 0.9% in water, BioXtra, suitable for cell culture
Sigma-Aldrich
Methanol, suitable for NMR (reference standard)
Supelco
Sodium hydroxide concentrate, 0.1 M NaOH in water (0.1N), Eluent concentrate for IC
Sigma-Aldrich
Imidazole, ReagentPlus®, 99%
Sigma-Aldrich
Sodium chloride-35Cl, 99 atom % 35Cl
Sigma-Aldrich
Sodium chloride solution, 0.85%
Supelco
Fructose, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Sodium chloride, random crystals, 99.9% trace metals basis
Millipore
D-(−)-Fructose, ≥99.0% (HPLC), suitable for microbiology
Supelco
Sodium hydroxide solution, 49-51% in water, eluent for IC
Sigma-Aldrich
D-(−)-Fructose, BioUltra, ≥99.0% (HPLC)
Sigma-Aldrich
Sodium hydroxide, BioUltra, suitable for luminescence, ≥98.0% (T), pellets
Sigma-Aldrich
Sodium hydroxide solution, BioUltra, Molecular Biology, 10 M in H2O
Sigma-Aldrich
D-(−)-Fructose, tested according to Ph. Eur.
Sigma-Aldrich
Sodium chloride, tested according to Ph. Eur.
Sigma-Aldrich
Sodium chloride solution, BioUltra, Molecular Biology, ~5 M in H2O
Supelco
Sodium chloride, reference material for titrimetry, certified by BAM, >99.5%
Supelco
Methanol, analytical standard
Sigma-Aldrich
Imidazole, BioUltra, Molecular Biology, ≥99.5% (GC)
Sigma-Aldrich
Imidazole, BioUltra, ≥99.5% (GC)
Sigma-Aldrich
Imidazole, puriss. p.a., ≥99.5% (GC)
Sigma-Aldrich
Sodium chloride, BioUltra, Molecular Biology, ≥99.5% (AT)
Sigma-Aldrich
3-Ethyl-2,4-pentanedione, mixture of tautomers, 98%
Sigma-Aldrich
Sodium chloride, AnhydroBeads, −10 mesh, 99.999% trace metals basis
Sigma-Aldrich
D-(−)-Fructose, ≥99% (HPLC), BioReagent, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
D-(−)-Fructose, ≥99% (HPLC)
Sigma-Aldrich
Sodium chloride solution, 5 M
Sigma-Aldrich
Sodium chloride, Molecular Biology, DNase, RNase, and protease, none detected, ≥99% (titration)
Sigma-Aldrich
Sodium chloride, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%