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Cloning and Characterization of a Weissella confusa Dextransucrase and Its Application in High Fibre Baking

机译:魏氏菌葡聚糖酶的克隆鉴定及其在高纤维烘烤中的应用

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摘要

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 14C-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.
机译:麦麸作为烘焙原料具有健康益处,但不利于面包的质地。通过微生物发酵生产右旋糖酐可以改善面包的体积和新鲜度,但是发酵过程中大量产酸可能会抵消这种影响。测试了麦麸中葡聚糖的酶促生产,以确定是否可以在不破坏面包质地的情况下将含葡聚糖的麸用于烘烤。对Weissella confusa VTT E-90392葡聚糖酶基因进行了测序,并在乳酸乳球菌中产生了带有His标签的葡聚糖酶Wc392-rDSR。使用 14 C-蔗糖放射性同位素和基于还原值的测定法对纯化的酶进行了表征,前者在酶解温度分别产生K m和V max值分别为14.7 mM和8.2μmol/(mg∙min)。最适pH为5.4。体外右旋糖酐产物的结构和大小类似于体内产生的右旋糖酐。使用Wc392-rDSR在6小时内在麦麸中生产了右旋糖酐(干重为8.1%)。含和不含右旋糖酐的麸皮用于小麦烘焙,添加量为20%。葡聚糖的存在改善了面包的柔软度,并中和了麸皮引起的体积损失,清楚地证明了在麸皮生物加工中使用葡聚糖转葡糖酶进行烘焙的潜力。

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