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《肠膜明串珠菌 NRRL B-1355 的葡糖基转移酶突变体》

Glucosyltransferase Mutants of Leuconostoc mesenteroides NRRL B-1355.

机构信息

Western Regional Research Center, U.S. Department of Agriculture, Albany, California 94710.

出版信息

Appl Environ Microbiol. 1994 Aug;60(8):2723-31. doi: 10.1128/aem.60.8.2723-2731.1994.

Abstract

Leuconostoc mesenteroides NRRL B-1355 produces dextrans and alternan from sucrose. Alternan is an unusual dextran-like polymer containing alternating alpha(1-->6)/alpha(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.

摘要

肠膜明串珠菌 NRRL B-1355 可利用蔗糖生产右旋糖酐和交替糖。交替糖是一种不寻常的葡聚糖样聚合物,含有交替的α(1-->6)/α(1-->3)糖苷键。利用紫外线和甲基磺酸乙酯对培养物进行诱变,然后在 10%蔗糖平板上选择菌落形态突变体。与亲代培养物相比,菌落形态变体在一种或多种葡糖基转移酶(GTF)和葡聚糖的产生方面发生了变化。通过测量葡聚糖产物对右旋糖酐酶消化的抗性、电泳和从蔗糖-麦芽糖混合物生成的麦芽糖受体制备物的高压液相色谱分析,对突变体进行了表征。一些突变体产生几乎纯的 L 型右旋糖酐,并且在十二烷基硫酸钠-丙烯酰胺凝胶上显示出单个主要的 GTF 带。其他突变体产生富含交替糖的葡聚糖。基于 GTF 的产生,解释了菌落形态特征(大小、光滑度和不透明度)和液体培养特性(10%蔗糖培养基中的团聚性、颜色和粘度)。检测到三个主要的 GTF 带。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b291/201715/e437ff2aeb0d/aem00025-0070-a.jpg

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