甜味蛋白植物

自上世纪70年代糖精被怀疑具有致癌作用以来,人们便开始搜寻各种无毒安全的新型甜味剂,其中植物甜味蛋白不仅甜度高、产生的热量少、可以防止肥胖;而且更重要的是这类甜味蛋白既无毒性,又不会使人产生龋齿,食用十分安全。因此,近年来已引起了人们广泛兴趣。迄今为止,人们已从下面6种植物中发现了这类高甜度的新型甜味剂。     

甜味蛋白研究进展

甜味蛋白是一类具有高甜度、低热卡、多功能的天然甜味剂。但要从几种古老的植物中提取甜味蛋白较为困难 ,且难以开发和利用。随着生物技术的发展 ,尤其是利用基因工程技术将甜味蛋白基因克隆到微生物细胞中 ,构建产甜味蛋白的基因工程菌 ,为商品化生产甜味蛋白开辟了一条快速而有效的新途径。     

甜味蛋白研究进展

甜味蛋白是一类具有高甜度,低热卡,多功能的天然甜味剂,但要从几种古老的植物中提取甜味蛋白较为困难,且难以开发利用,随着生物技术的发展,尤其是利用基因工程技术将甜味蛋白基因克隆到微生物细胞中,构建产甜味蛋白的基因工程菌,为商品化生长甜味蛋白开辟了一条快速而有效的新途径。     

植物甜味蛋白和矫味蛋白生物技术研究开发现状和前景

与蔗糖类甜味剂比较 ,植物甜味蛋白有很多优点 ,例如 :甜度高、热卡低、不引起龋齿以及可供糖尿病人食用等。迄今已知的植物甜味蛋白和矫味蛋白有 7种 ,即 :沙马汀 (Ｔｈａｕｍａｔｉｎ)、莫乃灵 (Ｍｏｎｅｌｌｉｎ)、马宾灵 (Ｍａｂｉｎｌｉｎ)、潘塔亭 (Ｐｅｎｔａｄｉｎ)、布拉齐因(Ｂｒａｚｚｅｉｎ)、库克灵 (Ｃｕｒｃｕｌｉｎ)和神秘果素 (Ｍｉｒａｃ ｕｌｉｎ)。这些植物甜味蛋白或矫味蛋白均来自热带雨林植物的果实。热带国家的居民常用其作为食物的增甜剂。近年来 ,在生物技术学家的努力下 ,又对这些甜味蛋白和矫味蛋白进行了克隆…     

奇甜蛋白基因在园艺作物中的表达

奇甜蛋白（thaumatin）是从非洲西部植物katemfe（Thaumatococcus daniellii Benth）中提取得到的几种关系相近的甜味蛋白的统称,其中最主要的为奇甜蛋白Ⅰ和奇甜蛋白Ⅱ。奇甜蛋白不仅甜度高,而且具有低热量、安全无毒以及不易诱发糖尿病等优点。因此,将奇甜蛋白基因转入园艺作物中并使之表达,用以提高可食部分的甜味,有其特别的研究意义。奇甜蛋白基因已先后在马铃薯、梨树、黄瓜、番茄等园艺作物得到表达,但仍有一些问题需要解决。现从奇甜蛋白基因的克隆、测序与表达,转基因果实的安全性检测,甜度的感官评价,甜味遗传特点以及奇甜蛋白抗真菌病害检验等几个方面综述了国内外研究进展,并对今后的研究提出了建议。     

甜味蛋白及其基因工程

甜味蛋白（ｔｈａｕｍａｔｉｎ）是从一种热带植物ＴｈａｕｍａｔｏｃｏｃｕｓｄｅｎｉｅｌｉＢｅｎｔｈ的胶质假种皮中分离出来的,该植物为竹芋科（Ｍａｒａｂｔａｃｅａｅ）的一种多年生灌木,广泛生长在西非的雨林中。甜味蛋白被认为是世界上已知的最甜的物质,大约２...     

Brazzein:一种耐热的甜味蛋白

Brazzein是从非洲西部野生植物Pentadiplandra brazzeana Baillon的果实中提取的一种甜味蛋白。Brazzein由54个氨基酸残基组成,Mr为6500,等电点为5,并且具有很好的热稳定性。本文着重概述Brazzein的结构与功能的关系,及其在基因工程中的研究前景。     

应乐果甜蛋白及其基因工程

应乐果甜蛋白（monellin)是一种分子量为10.7kD的超甜蛋白。由A,B两条链组成,其中A链44个氨基酸,B链50个氨基酸。分子内有5个反向平行的β折叠链和一个α螺旋。实验表明,Asp^B7可能是应乐果甜蛋白的甜味活性中心。此外,Cys^41,Ca^2 等对应乐果甜蛋白的甜味也会产生影响。研究人员把应乐果甜蛋白的单链类似物相断转入大肠杆菌,土豆,莴苣和酵母中,得到了具甜味,稳定性和耐受力强的表达产物。     

马槟榔甜味蛋白的研究 Ⅳ.稳定性和变性

通过对马槟榔甜昧蛋白Ⅰ和Ⅱ热变性试验,发现MaⅠ在80℃水浴中很快失甜味;圆二色谱证明其主链构象中的α螺旋几乎全部消失了。但Ma Ⅱ却能经受长时间保温而不失去甜味;电泳行为亦不发生改变。 在盐酸胍和尿素变性试验中发现,随变性剂浓度加大,蛋白质荧光发射光谱中色氨酸峰发生红移,直至蛋白质发生完全变性为止。以此为指标发现在酸性条件下,马槟榔甜蛋白更稳定;一般情况下3—5M的盐酸胍就足以使它变性;但即使是8M的尿素也不能使它完全变性。此和横向变性剂梯度聚丙烯酰胺凝胶电泳的结果是一致的。当脱掉8M尿素或—6M盐酸胍-MaⅠ溶液中的变性剂后,MaⅠ的甜味可恢复,其二级结构及电泳行为只发生微小的变化。 MaⅠ在SDS聚丙烯酰胺电泳分析中出现了一条双分子聚合体带,但将MaⅠ热变性、还原变性或羧甲基化等后,随MaⅠ甜味的消失,这条聚合体带也消失了。MaⅡ在任何情况下均不出现上述现象,因此认为这条带的形成与MaⅠ本身和甜味相关的一种特殊结构有关。     

植物甜蛋白Thaumatin研究进展

甜蛋白自 2 0世纪 70年代发现以来 ,一直倍受人们关注 ,而源于自然的Thaumatin是植物甜蛋白中的一种 ,它具有低热量、高甜度、安全无毒 ,并可降解为人体所需的氨基酸等多种优点 ,是一种新型甜味剂。在物质文化生活日益丰富的今天 ,人们越来越重视饮食的科学性 ,吃饱的同时更加关注所摄入食品的品质 ,无疑具多功能的非糖类物质 Thaumatin就是人们所需求的理想食品。因此 ,Thaumatin成为热门研究领域之一也就不足为怪了。1 　植物甜蛋白研究概况迄今为止 ,人们从多种植物中发现并分离出 7种甜味蛋白 [1 ]。更确切地说 ,其中 5种( Thaumatin,…     

植物凝集素的超级家族

凝集素是一类专一、可逆地和糖类结合的蛋白质,迄今已经分离纯化并测定了氨基酸序列的凝集素已有不少,一些凝集素以及它们与配体糖相互结合的复合物的高级结构也已经给出,许多工作已深入到基因水平．就目前已有的知识,说明植物凝集素是一个庞大的蛋白质家族．     

Protein redesign.

It has been demonstrated that, for a number of proteins, it is possible to dramatically alter the connectivities between elements of secondary structure. Remarkably large loop insertions are tolerated and many redesigns have generated proteins that successfully fold to stable, active structures. Some redesigns have been entirely the choice of the investigators, whereas others have incorporated a randomization and selection step to identify optimal sequences. These studies have provided basic guidelines for the rational manipulation of protein structure and stability, they have allowed the dissection of folding pathways and they have generated proteins with the potential for practical therapeutic applications.     

Lipid-transfer proteins: Tools for manipulating membrane lipids

Like other eukaryotic cells, plant cells contain proteins able to bind or to transfer lipids. Since they are able to facilitate movements of various phospholipids between membranes and are also capable of binding fatty acids or acyl-CoAs, they have been termed lipid-transfer proteins (LTP). LTPs are basic proteins containing 90 to 95 residues (molecular mass 9 kDa), eight of them being cysteines found in conserved locations. These proteins have been used to manipulate in vitro the lipid composition of isolated membranes either from plant or mammalian sources. In addition to purified LTPs, recombinant LTPs produced by genes expressed in microorganisms can be used for this purpose. Several genes coding for these proteins have been characterized in various plants with different patterns of expression. However, it remains to be investigated whether these recombinant proteins behave functionally as LTPs. The use of purified or recombinant LTPs is promising for the study of the effect of lipid composition on membrane functional properties.     

The leucine binding proteins of Escherichia coli as models for studying the relationships between protein structure and function

The genes encoding the leucine binding proteins in E coli have been cloned and their DNA sequences have been determined. One of the binding proteins (LIV-BP) binds leucine, isoleucine, valine, threonine, and alanine, whereas the other (LS-BP) binds only the D- and L-isomers of leucine. These proteins bind their solutes as they enter the periplasm, then interact with three membrane components, livH, livG, and livM, to achieve the translocation of the solute across the bacterial cell membrane. Another feature of the binding proteins is that they must be secreted into the periplasmic space where they carry out their function. The amino acid sequence of the two binding proteins is 80% homologous, indicating that they are the products of an ancestral gene duplication. Because of these characteristics of the leucine binding proteins, we are using them as models for studying the relationships between protein structure and function.     

Inter-organelle membrane contact sites: through a glass, darkly

Inter-organelle membrane contact sites are zones where heterologous membranes, usually the endoplasmic reticulum plus a partner organelle, come into close apposition. These sites are very poorly understood because so few of their components have been identified; however, it is clear that they are specialised for traffic of material and information between the two membranes. There have been recent advances in the study of lipid transfer proteins, such as ceramide transfer protein (CERT) and homologues of oxysterol binding protein (OSBP). Not only can these proteins carry lipids across the cytoplasm, but they have been found to target both the endoplasmic reticulum and a partnering organelle, and in some cases have been localised to membrane contact sites. Further work will be needed to test whether these lipid transfer proteins act when anchored at inter-organelle contact sites.     

Cold shock response and low temperature adaptation in psychrotrophic bacteria

Psychrotrophic bacteria are capable of developing over a wide temperature range and they can grow at temperatures close to or below freezing. This ability requires specific adaptative strategies in order to maintain membrane fluidity, the continuance of their metabolic activities, and protein synthesis at low temperature. A cold-shock response has been described in several psychrotrophic bacteria, which is somewhat different from that in mesophilic microorganisms: (i) the synthesis of housekeeping proteins is not repressed following temperature downshift and they are similarly expressed at optimal and low temperatures (ii) cold-shock proteins or Csps are synthesized, the number of which increases with the severity of the shock (iii) a second group of cold-induced proteins, i.e. the cold acclimation proteins or Caps, comparable with Csps are continuously synthesized during prolonged growth at low temperature. Homologues to CspA, the major cold-shock protein in E. coli, have been described in various psychrotrophs, but unlike their mesophilic counterparts, they belong to the group of Caps. Although they have been poorly studied, Caps are of particular importance since they differentiate psychrotrophs from mesophiles, and they are probably one of the key determinant that allow life at very low temperature.     

Saccharomyces cerevisiae cell cycle: cdc28 and the G1 cyclins.

Two families of cyclin-like proteins have been found in S. cerevisiae. The clb proteins are the mitotic cyclins. The cln proteins provide an essential function, are required for the G1/S transition, and appear to be rate-limiting for START, but have no obvious role elsewhere in the cycle. The cln proteins are unstable; they form complexes with cdc28; the complexes have protein kinase activity; and at least one of the clns oscillates in abundance through the cell cycle. The action of the cln cyclins at START suggests that they may be 'G1 cyclins'.     

Antifreeze proteins in winter rye

Six antifreeze proteins, which have the unique ability to adsorb onto the surface of ice and inhibit its growth, have been isolated from the apoplast of winter rye leaves where ice forms at subzero temperatures. The rye antifreeze proteins accumulate during cold acclimation and are similar to plant pathogenesis-related proteins, including two endoglucanase-like, two chitinase-like and two thaumatin-like proteins. Immunolocalization of the glucanase-like antifreeze proteins showed that they accumulate in mesophyll cell walls facing intercellular spaces, in pectinaceous regions between adjoining mestome sheath cells, in the secondary cell walls of xylem vessels and in epidermal cell walls. Because the rye antifreeze proteins are located in areas where they could be in contact with ice, they may function as a barrier to the propagation of ice or to inhibit the recrystallization of ice. Antifreeze proteins similar to pathogenesis-related proteins were also found to accumulate in closely-related plants within the Triticum group but not in freezing-tolerant dicotyledonous plants. In winter wheat, the accumulation of antifreeze proteins and the development of freezing tolerance are regulated by chromosome 5. Rye antifreeze proteins may have evolved from pathogenesis-related proteins, but they retain their catalytic activities and may play a dual role in increasing both freezing and disease resistance in overwintering plants.     

Properties of human mitochondrial ribosomes

Mammalian mitochondrial ribosomes (55S) differ unexpectedly from bacterial (70S) and cytoplasmic ribosomes (80S), as well as other kinds of mitochondrial ribosomes. Typical of mammalian mitochondrial ribosomes, the bovine mitochondrial ribosome has been developed as a model system for the study of human mitochondrial ribosomes, to address several questions related to the structure, function, biosynthesis and evolution of these interesting ribosomes. Bovine mitochondrial ribosomal proteins (MRPs) from each subunit have been identified and characterized with respect to individuality and electrophoretic properties, amino acid sequence, topographic disposition, RNA binding properties, evolutionary relationships and reaction with affinity probes of ribosomal functional domains. Several distinctive properties of these ribosomes are being elucidated, including their antibiotic susceptibility and composition. Human mitochondrial ribosomes lack several of the major RNA stem structures of bacterial ribosomes but they contain a correspondingly higher protein content (as many as 80 proteins), suggesting a model where proteins have replaced RNA structural elements during the evolution of these ribosomes. Despite their lower RNA content they are physically larger than bacterial ribosomes, because of the 'extra' proteins they contain. The extra proteins in mitochondrial ribosomes are 'new' in the sense that they are not homologous to proteins in bacterial or cytoplasmic ribosomes. Some of the new proteins appear to be bifunctional. All of the mammalian MRPs are encoded in nuclear genes (a separate set from those encoding cytoplasmic ribosomal proteins) which are evolving more rapidly than those encoding cytoplasmic ribosomal proteins. The MRPs are imported into mitochondria where they assemble coordinately with mitochondrially transcribed rRNAs into ribosomes that are responsible for translating the 13 mRNAs for essential proteins of the oxidative phosphorylation system.     

Changes in plasma membrane glycoproteins of rat spermatozoa during maturation in the epididymis

Glycoproteins on the plasma membrane of testicular and cauda epididymidal spermatozoa have been labeled with galactose oxidase/NaB [3H]4 and sodium metaperiodate/NaB[3H]4, followed by analysis on SDS polyacrylamide gels. The major glycoprotein labeling on testicular spermatozoa has a molecular weight 110,000 whereas on cauda epididymidal spermatozoa greater than 90% of the radio-label is incorporated into proteins of molecular weight 32,000. These 32,000-mol wt X proteins are homologous with proteins of similar molecular weight purified from the epididymal secretion and which have been shown previously to be synthesized in the caput epididymidis under hormonal control. Immunofluorescence revealed that the 32,000-mol wt proteins are present on the flagellum of mature but not immature spermatozoa and that they have a patchy distribution suggesting that they are mobile within the plane of the membrane. The membrane-bound 32,000-mol wt proteins possess hydrophobic domains as revealed by charge-shift electrophoresis and they also label with a lipophilic photoaffinity probe suggesting that they are in contact with the lipid bilayer. The evidence indicates that there is a considerable reorganization of the molecular structure of the plasma membrane of spermatozoa during maturation in the epididymis and that some of the changes are brought about by a direct interaction with epididymal secretory proteins.