Laboratoire de Biologie appliquée à l'Agroalimentaire et à l'Environnement, Institut Universitaire de Technologie, Université Paul Sabatier, Auch, France.
FEMS Microbiol Lett. 2010 Oct;311(1):18-26. doi: 10.1111/j.1574-6968.2010.02067.x. Epub 2010 Aug 16.
The study of exopolysaccharide production by heterofermentative sourdough lactic acid bacteria has shown that Weissella strains isolated from sourdoughs produce linear dextrans containing α-(1→6) glucose residues with few α-(1→3) linkages from sucrose. In this study, several dextran-producing strains, Weissella cibaria and Weissella confusa, isolated from sourdough, were characterized according to carbohydrate fermentation, repetitive element-PCR fingerprinting using (GTG)(5) primers and glucansucrase activity (soluble or cell-associated). This study reports, for the first time, the characterization of dextransucrase from Weissella strains using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and in situ polymer production (after incubation with sucrose) from enzymatic fractions harvested from both sucrose and glucose culture media. Results demonstrate that dextransucrase activity was mainly soluble and associated with a constitutive 180-kDa protein. In addition, microsequencing of the active dextransucrase from W. cibaria LBAE-K39 allowed the design of specific primers that could detect the presence of glucansucrase encoding genes similar to GTFKg3 of Lactobacillus fermentum Kg3 and to DSRWC of W. cibaria CMU. This study hence indicates that sourdough Weissella strains synthesize original dextransucrase.
异戊糖发酵型酸面团乳酸菌胞外多糖生产的研究表明,从酸面团中分离出的魏斯氏菌(Weissella)菌株利用蔗糖产生含有少量α-(1→3) 键的线性葡聚糖,其葡萄糖残基通过α-(1→6)键连接。本研究根据碳水化合物发酵、(GTG)(5)引物的重复元件-PCR 指纹图谱和葡聚糖蔗糖酶活性(可溶性或细胞结合)对几种产葡聚糖的菌株(魏斯氏菌 cibaria 和魏斯氏菌 confusa)进行了表征。本研究首次报道了使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和原位聚合(在与蔗糖孵育后)从蔗糖和葡萄糖培养基中收获的酶级分中对魏斯氏菌菌株的葡聚糖蔗糖酶进行表征。结果表明,葡聚糖蔗糖酶活性主要是可溶性的,并与一种组成型 180kDa 蛋白相关。此外,对 W. cibaria LBAE-K39 的活性葡聚糖蔗糖酶进行微测序,设计了特异性引物,能够检测到类似于 L. fermentum Kg3 的 GTFKg3 和 W. cibaria CMU 的 DSRWC 的葡聚糖蔗糖酶编码基因的存在。因此,本研究表明酸面团魏斯氏菌菌株合成了原始的葡聚糖蔗糖酶。