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Glucosyltransferase Mutants of Leuconostoc mesenteroides NRRL B-1355

机译:mesenteroides NRRL B-1355的葡糖基转移酶突变体

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摘要

Leuconostoc mesenteroides NRRL B-1355 produces dextrans and alternan from sucrose. Alternan is an unusual dextran-like polymer containing alternating α(1→6)/α(1→3) glucosidic bonds. Cultures were mutagenized with UV and ethyl methanesulfonate, and colony morphology mutants were selected on 10% sucrose plates. Colony morphology variants exhibited changes from parent cultures in the production of one or more glucosyltransferases (GTFs) and glucans. Mutants were characterized by measuring resistance of glucan products to dextranase digestion, by electrophoresis, and by high-pressure liquid chromatography of maltose acceptor products generated from sucrose-maltose mixtures. Some mutants produced almost pure fraction L dextran, and cultures exhibited a single principal GTF band on sodium dodecyl sulfate-acrylamide gels. Other mutants produced glucans enriched for alternan. Colony morphology characteristics (size, smoothness, and opacity) and liquid culture properties (clumpiness, color, and viscosity in 10% sucrose medium) were explained on the basis of GTF production. Three principal GTF bands were detected.
机译:Mesenteroides NRRL B-1355肠球菌由蔗糖产生葡聚糖和交替糖。 Alternan是一种不寻常的葡聚糖样聚合物,包含交替的α(1→6)/α(1→3)糖苷键。用UV和甲烷磺酸乙酯诱变培养物,并在10%蔗糖平板上选择菌落形态突变体。菌落形态变异在一种或多种葡糖基转移酶(GTF)和葡聚糖的生产中表现出与亲本培养有关的变化。通过测量葡聚糖产物对葡聚糖酶消化的抗性,电泳和通过蔗糖-麦芽糖混合物产生的麦芽糖受体产物的高压液相色谱来表征突变体。一些突变体产生几乎纯净的级分L葡聚糖,并且培养物在十二烷基硫酸钠-丙烯酰胺凝胶上显示出单个主要GTF带。其他突变体产生的葡聚糖富含交替糖。根据GTF的产生情况,解释了菌落的形态特征(大小,光滑度和不透明性)和液体培养特性(在10%蔗糖培养基中的结块性,颜色和粘度)。检测到三个主要的GTF带。

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