无机溶剂法制β-环糊精的工艺研究

对一种无机溶剂酶催化生产β-环糊精的工艺进行研究,并采用响应面实验法确定了生产β-环糊精的适宜工艺条件,为其开发研究应用提供参考。实验结果表明,生产争环糊精的最佳条件是选取木薯淀粉作为底物,不经过预处理,环状糊精葡萄糖基转移酶添加量是4u/g,CaCl2的浓度为30mmol/L,pH7．88,温度63．67℃,反应时间17．24h,产率可达24．02％。生产工艺和传统工艺相比,产率明显增高,提高了生产效率。     

环糊精系列综述之一 环糊精葡萄糖基转移酶的筛选及其定向改造

环糊精因其特殊性质得到了食品、化妆品、医药等行业的青睐,全球消费量逐年稳步增加。环糊精的巨大需求也使其生产中所必需的环糊精葡萄糖基转移酶(CGT酶,EC 2.4.1.19)成为研究热点。目前,对该酶的研究多集中在作用机制、产物专一性机理等,鲜见从菌株筛选出发的系统性报道。因此,作者从野生CGT酶菌株的筛选出发,立足未来环糊精工业的商业诉求,综述了优质CGT酶获取的多种途径。同时结合国内外对CGT酶的大量研究工作以及我国的资源优势,对该领域未来的研究提出更高的期望。     

Improvement of cyclodextrin glucanotransferase as an antistaling enzyme by error-prone PCR

In an effort to improve the properties of cyclodextrin glucanotransferase (CGTase) as an antistaling enzyme, error-prone PCR was used to introduce random mutations into a CGTase cloned from alkalophilic Bacillus sp. I-5 (CGTase I-5). A mutant CGTase[3-18] with the three mutations M234T, F259I and V591A was selected by agar plate assay. Sequence alignment of various CGTases indicated that M234 and F259 are located in the vicinity of the catalytic sites of the enzyme and V591 in the starch binding domain E. The cyclization activity of CGTase[3-18] was dramatically decreased by 10-fold, while the hydrolyzing activity was increased by up to 15-fold. These mutations near subsite +1 (M234T) and at subsite +2 (F259I) are likely to alter the enzyme activity in a concerted manner, promoting hydrolysis of substrate while retarding cyclization. The addition of CGTase[3-18] reduced the retrogradation rate of bread by as much as did the commercial antistaling enzyme Novamyl during 7-day storage at 4 degrees C. No cyclodextrin (CD) was detected in bread treated with CGTase[3-18], whereas 21 mg of CD per 10 g of bread was produced in bread treated with wild-type CGTase.     

环糊精制备过程的催化机制及其影响因素

环糊精主要是由环糊精葡萄糖基转移酶(CGT酶,EC 2.4.1.19)作用淀粉制备而得。除受CGT酶的来源及种类的影响之外,影响环糊精产率及产物特异性的因素还有很多,诸如酶作用时间、作用温度、pH、加酶量、添加物、底物种类、底物浓度、原料预处理程度等;此外,这些因素还将影响环糊精的制备工艺过程。然而,目前尚未有环糊精制备过程影响因素的系统报道。作者从环糊精糖基转移酶的催化机理分析入手,阐述影响环糊精产率及产物特异性的影响因素,同时结合环糊精的制备工艺,对环糊精制备相关研究进展进行综述,以期为在环糊精制备相关领域开展研究提供参考。     

Rational mutagenesis of cyclodextrin glucanotransferase at the calcium binding regions for enhancement of thermostability

Studies related to the engineering of calcium binding sites of CGTase are limited. The calcium binding regions that are known for thermostability function were subjected to site-directed mutagenesis in this study. The starting gene-protein is a variant of CGTase Bacillus sp. G1, reported earlier and denoted as "parent CGTase" herein. Four CGTase variants (S182G, S182E, N132R and N28R) were constructed. The two variants with a mutation at residue 182, located adjacent to the Ca-I site and the active site cleft, possessed an enhanced thermostability characteristic. The activity half-life of variant S182G at 60 °C was increased to 94 min, while the parent CGTase was only 22 min. This improvement may be attributed to the formation of a shorter α-helix and the alleviation of unfavorable steric strains by glycine at the corresponding region. For the variant S182E, an extra ionic interaction at the A/B domain interface increased the half-life to 31 min, yet it reduced CGTase activity. The introduction of an ionic interaction at the Ca-I site via the mutation N132R disrupted CGTase catalytic activity. Conversely, the variant N28R, which has an additional ionic interaction at the Ca-II site, displayed increased cyclization activity. However, thermostability was not affected.     

新奇环状淀粉——大环糊精的最新研究进展

大环糊精是一类由9个以上葡萄糖基组成的环状糊精混合物总称,由于其特异的分子结构和性质,具有广泛的工业应用前景,最近10年在国际上引起了广泛的关注和研究。分析了在大环糊精的合成、分离纯化及其结构和应用等方面的最新研究进展,指出了存在的问题和下一步的研究方向,并分析了其潜在的工业应用前景。     

酶法改质甜菊糖的研究

研究环糊精葡糖基转移酶(CGTase)酶法改质甜菊糖的各种影响因素,确定适宜的反应条件,结果如下:甜菊糖浓度为15%,淀粉∶甜菊糖为2∶1,CGTase的添加量为0.3 mL,在65℃条件下反应30 h。     

环糊精的性能、生产及其在食品工业中的应用

本文介绍了环糊精的结构、性能、生产方法及其在食品工业中的应用等。