高产洛伐他汀红曲菌的选育及其在红腐乳中的应用

硫酸软骨素酶（ChSase）是一类能将硫酸软骨素（CS）催化裂解为小分子多糖的酶,根据结合的底物种类可分为ChSase ABC,ChSase AC,ChSase B和ChSase C。将普通变形杆菌（Proteus vulgaris）KCTC 2579的硫酸软骨素酶ABC I（ChSase ABC I）与麦芽糖结合蛋白（MBP）融合表达,构建了pMAL-c2x-ChSase ABC I重组载体,并成功在大肠杆菌（Escherichia coli）BL21(DE3)宿主中高效表达。将麦芽糖结合蛋白-硫酸软骨素酶（MBP-ChSase）ABC I固定在聚苯胺载体上,制备生物传感器,并利用循环伏安法对其进行检测。结果表明,MBP-ChSase ABC I酶活和比酶活分别为3 180 IU/L发酵液和76 IU/mg蛋白。选择硫酸软骨素A（CS A）作为酶的反应底物,在最适反应条件下,反应的峰值电流与CS A在浓度0.1 nmol/L～0.1 μmol/L范围内呈线性关系。该生物传感器对底物反应迅速,具有较高的灵敏度,可以进行有效检测。     

模拟肠液培养基中壳寡糖对益生菌生长的影响

文章探究了模拟肠液培养基中壳寡糖对益生菌生长的影响,为益生菌、壳寡糖合生元的开发提供理论依据。将益生菌接种至含不同浓度壳寡糖的模拟肠液培养基中培养,通过测定菌体密度(△OD₆₀₀)、活菌数的变化,测定△OD₂₆₀的变化以及用扫描电子显微镜表征细胞膜的完整性,明确模拟肠液培养基中壳寡糖对益生菌生长、细胞膜完整性的影响。结果显示8株专利益生菌都可以在模拟肠液培养基中正常生长,都不能以壳寡糖为唯一碳源生长。自然培养过程模拟肠液培养基中壳寡糖抑制8株专利益生菌的生长;壳寡糖处理导致益生菌细胞膜表面塌陷及褶皱,破坏了益生菌细胞膜的完整性。偏碱性模拟肠液培养基中壳寡糖对8株专利益生菌的生长没有统一的抑制作用。偏碱性模拟肠液环境下,壳寡糖对益生菌生长的抑制作用较弱,或没有抑制作用,可以将益生乳酸菌与壳寡糖复配开发合生元,用于调节肠道微生态。     

酶法制备透明质酸寡糖及其透皮吸收活性研究

为研究透明质酸寡糖的透皮吸收活性,采用透明质酸裂解酶对透明质酸进行酶法降解,制备透明质酸寡糖,通过红外光谱与质谱对透明质酸寡糖进行分析,并对其吸湿性能、体表保湿性能和透皮吸收性能进行评价.结果显示,透明质酸裂解酶酶解制备透明质酸寡糖的最适底物浓度为1％,最适加酶量为3.0 U/mg,酶解制备透明质酸寡糖基本没有基团脱落...     

玉米醇溶蛋白酶法糖基化修饰及产物性质研究

以转谷氨酰胺酶为催化剂,壳寡糖为酰基受体,通过糖基化反应修饰玉米醇溶蛋白。以修饰产物中氨基葡萄糖的导入量为指标,采用单因素试验优化糖基化反应条件,并对修饰产物的部分功能性质进行研究。结果表明:壳寡糖糖基化修饰玉米醇溶蛋白的最优条件为:反应初始pH 7.7,温度37℃,底物质量分数3%,酰基供体与酰基受体的物质的量比1∶3,加酶量60 U/g蛋白,反应时间8 h。在此条件下,壳寡糖的接入量最大,为101.47 mg氨基葡萄糖/g蛋白。与玉米醇溶蛋白和交联玉米醇溶蛋白相比,糖基化修饰玉米醇溶蛋白的溶解性和体外消化性均明显改善。     

不同连接肽在硫酸软骨素酶 ABC I重组表达中的应用研究

硫酸软骨素酶ABC I（ChSase ABC I）是一类能够将硫酸软骨素、软骨素、透明质酸等糖胺多糖降解为寡糖及不饱和二糖的裂解酶。该研究将硫酸软骨素酶ABC I基因与麦芽糖结合蛋白（MBP）基因分别用FFFFF和RRRRR两种连接肽连接，并克隆到pMAL-c2X载体上，在大肠杆菌（Escherichia coli）BL21(DE3)高效表达。结果表明，重组酶MBP-FFFFF-ChSase ABC I的酶活和比酶活分别为716.5 IU/L发酵液和1.15 IU/mg蛋白，重组酶MBP-RRRRR-ChSase ABC I的酶活和比酶活分别为741.0 IU/L发酵液和1.13 IU/mg蛋白。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）结果表明，重组酶MBP-FFFFF-ChSase ABC I和MBP-RRRRR-ChSase ABC I的分子质量均为130 kDa，并且都为可溶性蛋白。     

非消化性寡糖对两株益生菌体外增殖的影响

测定干酪乳杆菌(Lactobacilluscasei)LC2W和植物乳杆菌(Lactobacillusplantarum)ST-III在以非消化性寡糖为唯一碳源的MRS液体培养液中的吸光值,以及在添加非消化性寡糖的脱脂牛乳中发酵后,发酵乳中益生菌菌落总数、乳酸含量和pH,研究非消化性寡糖对L.caseiLC2W和L.plantarumST-III体外增殖的影响。结果显示,低聚半乳糖和菊粉可以明显促进L.caseiLC2W的体外增殖(P0.01),其最佳添加浓度为3%;而对L.plantarumST-III体外增殖促进作用最显著的是棉子糖和菊粉(P0.01),最佳添加浓度为2%。研究表明不同益生菌菌株对非消化性寡糖发挥体外增殖效应具有选择性,研究结果可为益生菌产品的开发提供技术基础。     

后生元的功能及其应用

随着人们生活质量的提高,对疾病及健康的关注度也随之提高。人们不再将关注点放于治疗疾病的相关医疗器械与药物成分,而是逐渐对起着预防、改善或辅助性缓解作用的新兴物质、成分或加工产品进行着重了解。这类物质对机体的作用更加侧重于通过定期的外部摄入来维持生理功能的运转或或促进健康。可外部摄入且有益于机体的物质最基本的要求是食用安全,而无副作用、多种类、多元化将、高效方便将是人们未来关注的新标签。定期摄入具有安全认证的益生菌产品,被认为是改善人体健康状况以及缓解疾病的一种较为安全且普遍的方法,而益生菌存活率低,发挥作用要求较高,使得其在大规模制备、长时间运输储存并保证其作用特性上存在一定的困难。到目前为止,各种保证益生菌高活性的制备、储存方法也在被不断地研究。近年来,伴随人们对益生菌研究的深入,已发现由益生菌通过不同制备方法所得的后生元也可对机体产生积极影响,并具备益生菌产品所不具有的稳定且储运方便等特点,或将成为未来改善机体状况或增强机体功能的新方向。本文综述后生元对机体的各种功能,其相关产品在各领域的应用情况。     

分子动力学模拟研究壳寡糖与脂肪酶相互作用机制

壳聚糖及其衍生物在酶固定化领域有着广泛的应用.了解酶与壳聚糖衍生物在分子水平上的相互作用机制,对相关生物催化剂的设计和应用具有重要意义.本研究采用分子动力学模拟方法研究南极假丝酵母脂肪酶B(CaLB)与壳寡糖(OCTS)的相互作用,并对脂肪酶与催化底物进行分子对接研究脂肪酶催化活性中心和底物结合构象和结合亲和力.结果表...     

益生菌的功能及安全性评价

正长期以来,益生菌产品被认为是有益且安全的,已经被广泛应用于食品加工生产、医药治疗、微生物制剂等领域。益生菌的许多有益功能已被科学实验证明,包括改善胃肠道功能、调节机体免疫功能、降低血清胆固醇、促进营养成分吸收和去除重金属等。本文综述了益生菌的主要生理功能,并对其安全性评价方法进行了总结。益生菌是一种通过摄入足够的数量,能够对宿主起到积极作用的活的微生物。其中大部分是乳酸菌,包括     

组成型壳聚糖酶制备壳寡糖酶解条件的优化

主要对组成型壳聚糖酶降解壳聚糖制备低壳寡糖的工艺进行了研究。以酶解产物壳寡糖的产量(mg)为指标,酶解pH、酶解温度(℃)、底物浓度(%)、酶解时间(min)为自变量,在单因素试验的基础上,通过正交实验确定了最佳酶解工艺为:酶解pH 7.0、酶解温度50℃、酶解时间90 min、底物浓度1%,此时壳寡糖产含量为5.476 mg。     

日本益生菌、益生元市场与应用概况

为了大力推广益生菌和益生元,介绍了日本益生菌制品、益生元、灭活细胞粉及对它们的管理情况。其中,日本厚生劳动省批准了65种特定保健食品,允许使用了近20种菌株．同时,各种功能性低聚糖已广泛使用在400多种食品、保健食品以及宠物饲料中,并说明了除益生菌活菌体外,益生菌或肠道正常菌群的灭活细胞,在大量存在下也可和活菌体一样具有免疫增强作用和发挥益生元的功能的作用。     

Non-digestible oligosaccharides with prebiotic properties

The search for functional foods or functional food ingredients, i.e. foods or food ingredients that can enhance health, is beyond any doubt one of the leading trends in today's food industry. In this context, probiotics, i.e. living microbial food supplements, and prebiotics, i.e. non-digestible food ingredients, receive much attention. Both popular concepts target the gastrointestinal microbiota. While in the Western world, intake of probiotics has been recommended for long, prebiotics in general, and non-digestible oligosaccharides in particular, have only recently received attention. This review deals with production and characterization of non-digestible oligosaccharides and focuses on their role in promoting health and treating diseases. Attention is paid to the effects of non-digestible oligosaccharides on constipation, mineral absorption, lipid metabolism, cancer prevention, hepatic encephalopathy, glycemia/insulinemia, and immunomodulation.     

6种低聚糖对肠道益生菌生长情况的影响及代谢产物分析

该研究考察了水苏糖、低聚半乳糖、低聚果糖、菊粉、低聚木糖、抗性糊精等6种低聚糖对9种肠道益生菌的体外增殖作用,评价了不同低聚糖对不同益生菌的促生长和促产酸能力,并分析该54个发酵体系中乙酸和乳酸的最大产生量.结果 表明,6种低聚糖对各益生菌的促生长和促产酸能力不同,以水苏糖和低聚半乳糖的效果最为显著,表现出广谱的代谢能...     

Selection of probiotics and prebiotics for synbiotics and confirmation of their in vivo effectiveness

Influence of fructan-type oligosaccharides (as prebiotics) on growth and acidifying activity of Bifidobacterium strains (as probiotics) was studied in vitro, using minimal nutrition media. The selected synbiotic pairs of stimulated bifidobacteria strains and oligosaccharides enhancing their growth were studied in vivo to determine the effect of probiotics, prebiotics and synbiotics on microecology of gut. Daily, >10⁹ live cells of bifidobacteria strains and/or 5% (w/w) of oligofructose in the diet were orally administered to Wistar rats. After a 14-day feeding experiment, the numbers of faecal bifidobacteria, aerobic and facultatively anaerobic bacteria, coliforms, and spores of anaerobic and aerobic bacteria were determined using microbiological methods. In vitro studies showed that the majority of Bifidobacterium species utilised FOS and low-polymerised inulins, but only 18 out of 30 strains tested (mostly of B. longum and B. animalis species) were stimulated. Incorporation of oligofructose into the diet stimulated the proliferation of faecal bifidobacteria by 1.6 log cfu/g in comparison to the control. B. longum KN29.1 and KNA1 selected in vitro, slightly increased the faecal bifidobacteria live cell number by about 0.6 log cfu/g under in vivo conditions, whereas B. animalis KSp4 was not effective. But administration of bifidobacteria together with the prebiotic (as synbiotics) improved the bifidogenic effect by 1.4 log cfu/g of faeces. Supplementation of diet had almost no effect on the other determined groups of gut microflora. The obtained results showed the selective stimulation of faecal bifidobacteria by probiotics, prebiotic and synbiotics, a great bifidogenic effectiveness of oligofructose, confirmed proper selection of synbiotics and showed their higher effectiveness in relation to probiotics.     

低聚果糖和低聚半乳糖的肠道益生功能研究

益生元低聚糖是一种可被选择性发酵并调节肠道中有益菌群的组成和活性的食物成分。其中功能性低聚糖在人体肠道内可被双歧杆菌和乳酸杆菌等肠道益生菌利用,进而促进肠道益生菌的生长和增殖。采用传统体外培养与体内试验方法,以调节婴儿肠道有益菌群为目的,评价低聚果糖和低聚半乳糖的益生功能。结果表明:两种功能性低聚糖均对肠道有益菌有较好的增殖作用;能有效调节肠道菌群失衡的情况;动物试验中对照组与试验组差别较大。     

海洋寡糖益生菌微胶囊的制备和体外评估及其对动物肠道菌群的调节作用

为解决益生菌不耐低pH值、不耐氧的缺点,采用微胶囊包埋技术,以海藻酸钠为壁材用乳化法制备长双歧杆菌藻酸盐微胶囊。将基础藻酸盐微胶囊外包壳寡糖（chitosan oligosaccharides,COS）或在藻酸盐壁材中添加藻酸盐寡糖（alginate oligosaccharides,AOS）分别制备两种海洋寡糖微胶囊：COS-微胶囊和AOS-COS-微胶囊。体外实验表明：两种海洋寡糖微胶囊均可以显著提高长双歧杆菌在模拟消化液处理后或在4 ℃贮藏期内的存活率。AOS-COS-微胶囊在模拟胃液处理后仍能保持106 CFU/g以上的活菌数。在连续的模拟消化液处理后,AOS-COS-微胶囊中的活菌量达到基础微胶囊中的约1 000 倍。两种海洋寡糖微胶囊在4 ℃贮藏28 d后仍均能保持108 CFU/g以上的活菌数。体内实验表明：相比较未包埋的长双歧杆菌或基础微胶囊,海洋寡糖微胶囊可以显著提高动物肠道菌群中益生菌的含量,同时降低条件致病菌的含量,具有最佳的调节肠道菌群效果。因此,海洋寡糖益生菌微胶囊产品将是一种有巨大应用前景的新型功能性益生菌食品。     

Galactooligosaccharides: novel components of designer foods

Since introduction of functional foods, commercialization of the traditionally used probiotics has ushered in more followers into the new fraternity of sophisticated, health-conscious consumers. In 1995, this was followed by the first introduction of prebiotics. Prebiotics are defined as "a non-digestible feed supplement, beneficially affecting the host by selectively stimulating growth and/or activity in one or a limited number of bacteria in the colon." The number of new product introductions with prebiotics has steeply increased over the last few years. Paradoxically, probiotics have limited applications as these cannot be used in wide range of food products because of their viability issue. Fortunately, prebiotics do not suffer from any such constraint and can be used in a wide range of food products. Probiotics do not have a long shelf life in their active form. In most cases, refrigeration is required to maintain the shelf life. While probiotics are predominantly used in fermented dairy products, the use of prebiotics has expanded into other food categories. Prebiotics have successfully been incorporated in a wide variety of human food products such as baked goods, sweeteners, yoghurts, nutrition bars, and meal replacement shakes. For instance, the introduction of galacto-oligosaccharides (GOS) into baby foods has been very successful. GOS, which are identical to the human milk oligosaccharides, has emerged with strong clinical support for both digestive and immune health. Various aspects related to GOS such as types and functions of functional food constituents with special reference to GOS, their role as prebiotics, and enhanced industrial production through microbial intervention are dealt in this review.     

Effects of oligosaccharides on phase transition temperatures and rheological characteristics of waxy rice starch dispersion

BACKGROUND: The creation of starch‐based foods incorporated with functional ingredients such as probiotics is of great current interest in the food industry. This study aimed to investigate the effects of prebiotic oligosaccharides on the phase transition temperatures and rheological characteristics of waxy rice starch dispersions. Four oligosaccharides were applied to the rice starch dispersions: chitooligosaccharides, fructooligosaccharides, isomaltooligosaccharides and xylooligosaccharides. RESULTS: The addition of 125 g kg^?1 oligosaccharides elevated the onset and peak temperatures for gelatinisation of 200–400 g kg^?1 waxy rice starch dispersions. The temperature of the storage modulus (G′) for gelatinisation increased markedly on adding fructooligosaccharides to 200–300 g kg^?1 waxy rice starch. For gelatinisation of 300 g kg^?1 rice starch dispersion the effectiveness of the oligosaccharides in changing the above parameters was as follows: chitooligosaccharides > fructooligosaccharides > isomaltooligosaccharides > xylooligosaccharides. Moreover, their effectiveness was dependent on the amylose content, as illustrated by comparing waxy and non‐waxy rice starches (amylose contents 9–256 g kg^?1). Importantly, the logarithmic G′₉₅ change was linearly and negatively correlated with amylose content. CONCLUSION: The results suggest that oligosaccharide‐containing rice starch dispersions may potentially be used for the formulation of oligosaccharide‐containing starchy functional foods owing to the rheological changes of these starch dispersions. Copyright © 2011 Society of Chemical Industry     

Synthesis of β‐Galactooligosaccharides from Lactose Using Microbial β‐Galactosidases

Abstract: Galactooligosaccharides (GOSs) are nondigestible oligosaccharides and are comprised of 2 to 20 molecules of galactose and 1 molecule of glucose. They are recognized as important prebiotics for their stimulation of the proliferation of intestinal lactic acid bacteria and bifidobacteria. Therefore, they beneficially affect the host by selectively stimulating the growth and/or activity of a limited number of gastrointestinal microbes (probiotics) that confer health benefits. Prebiotics and probiotics have only recently been recognized as contributors to human health. A GOS can be produced by a series of enzymatic reactions catalyzed by β‐galactosidase, where the glycosyl group of one or more D‐galactosyl units is transferred onto the D‐galactose moiety of lactose, in a process known as transgalactosylation. Microbes can be used as a source for the β‐galactosidase enzyme or as agents to produce GOS molecules. Commercial β‐galactosidase enzymes also do have a great potential for their use in GOS synthesis. These transgalactosyl reactions, which could find useful application in the dairy as well as the larger food industry, have not been fully exploited. A better understanding of the enzyme reaction as well as improved analytical techniques for GOS measurements are important in achieving this worthwhile objective.