Department of Biotechnology, Indian Institute of Technology Guwahati, Guwahati, 781 039 Assam India.
Indian J Microbiol. 2007 Sep;47(3):197-206. doi: 10.1007/s12088-007-0040-8. Epub 2007 Oct 4.
The enzyme dextransucrase (sucrose:1, 6-α-D-glucan 6-α-glucosyltransferase, EC 2.4.1.5) catalyses the synthesis of exopolysaccharide, dextran from sucrose. This class of polysaccharide has been extensively exploited in pharmaceutical industry as blood volume expander, as stabiliser in food industry and as a chromatographic medium in fine chemical industry because of their nonionic nature and stability. Majority of the dextrans are synthesized from sucrose by dextransucrase secreted mainly by bacteria belonging to genera Leuconostoc, Streptococcus and Lactobacillus. Bulk of the information on purification of extracellular dextransucrase has been generated from Leuconostoc species. Various methods such as precipitation by ammonium sulphate, ethanol or polyethylene glycol, phase partitioning, ultrafiltration and chromatography have been used to purify the enzyme. Purification of dextransucrase is rendered difficult by the presence of viscous dextran in the medium. However, processes like ultra-filtration, salt and PEG precipitation, chromatography and phase partitioning have been standardized and successfully used for higher scale purification of the enzyme. A recombinant dextransucrase from Leuconostoc mesenteroides B-512F with a histidine tag has been expressed in E. coli cells and purifi ed by immobilized metal ion chromatography. This review reports the available information on purifi cation methods of dextransucrase from Leuconostoc mesenteroides strains.
酶葡聚糖蔗糖酶(蔗糖:1,6-α-D-葡聚糖 6-α-葡糖苷基转移酶,EC 2.4.1.5)催化多糖,葡聚糖的合成来自蔗糖。由于其非离子性质和稳定性,该类多糖已在制药工业中广泛用作血容量扩张剂,在食品工业中用作稳定剂,并在精细化工中用作色谱介质。大多数葡聚糖是由主要由肠膜明串珠菌、链球菌和乳杆菌属细菌分泌的葡聚糖蔗糖酶从蔗糖合成的。关于胞外葡聚糖蔗糖酶的纯化的大量信息是从肠膜明串珠菌属中获得的。已经使用了各种方法,例如硫酸铵、乙醇或聚乙二醇沉淀、相分配、超滤和色谱法来纯化酶。由于培养基中存在粘性葡聚糖,因此葡聚糖蔗糖酶的纯化变得困难。然而,超滤、盐和 PEG 沉淀、色谱和相分配等过程已经标准化,并成功用于该酶的更高规模纯化。来自肠膜明串珠菌 B-512F 的带有组氨酸标签的重组葡聚糖蔗糖酶已在大肠杆菌细胞中表达,并通过固定化金属离子色谱法进行纯化。本综述报告了来自肠膜明串珠菌菌株的葡聚糖蔗糖酶的纯化方法的现有信息。