Human intestinal maltase-glucoamylase: crystal structure of the N-terminal catalytic subunit and basis of inhibition and substrate specificity

Human maltase-glucoamylase (MGAM) is one of the two enzymes responsible for catalyzing the last glucose-releasing step in starch digestion. MGAM is anchored to the small-intestinal brush-border epithelial cells and contains two homologous glycosyl hydrolase family 31 catalytic subunits: an N-terminal subunit (NtMGAM) found near the membrane-bound end and a C-terminal luminal subunit (CtMGAM). In this study, we report the crystal structure of the human NtMGAM subunit in its apo form (to 2.0 Å) and in complex with acarbose (to 1.9 Å). Structural analysis of the NtMGAM-acarbose complex reveals that acarbose is bound to the NtMGAM active site primarily through side-chain interactions with its acarvosine unit, and almost no interactions are made with its glycone rings. These observations, along with results from kinetic studies, suggest that the NtMGAM active site contains two primary sugar subsites and that NtMGAM and CtMGAM differ in their substrate specificities despite their structural relationship. Additional sequence analysis of the CtMGAM subunit suggests several features that could explain the higher affinity of the CtMGAM subunit for longer maltose oligosaccharides. The results provide a structural basis for the complementary roles of these glycosyl hydrolase family 31 subunits in the bioprocessing of complex starch structures into glucose.     

叶绿素荧光动力学参数的意义及讨论

叶绿素荧光动力学技术被称为研究植物光合功能的快速、无损伤探针。但其参数众多，且名称及在参数的生物学意义解释上存在不规范和混乱现象。本文通过对这些问题的讨论旨在引起使用者的注意，并探讨正确使用这些参数的途径     

Microbial sucrose isomerases: producing organisms, genes and enzymes

Sucrose isomerase (SI) activity is used industrially for the conversion of sucrose into isomers, particularly isomaltulose or trehalulose, which have properties advantageous over sucrose for some food uses. All of the known microbial SIs are TIM barrel proteins that convert sucrose without need for any cofactors, with varying kinetics and product specificities. The current analysis was undertaken to bridge key gaps between the information in patents and scientific publications about the microbes and enzymes useful for sucrose isomer production.This analysis shows that microbial SIs can be considered in 5 structural classes with corresponding functional distinctions that broadly align with the taxonomic differences between producing organisms. The most widely used bacterial strain for industrial production of isomaltulose, widely referred to as “Protaminobacter rubrum” CBS 574.77, is identified as Serratia plymuthica. The strain producing the most structurally divergent SI, with a high product specificity for trehalulose, widely referred to as “Pseudomonas mesoacidophila” MX-45, is identified as Rhizobium sp.Each tested SI-producer is shown to have a single SI gene and enzyme, so the properties reported previously for the isolated proteins can reasonably be associated with the products of the genes subsequently cloned from the same isolates and SI classes. Some natural isolates with potent SI activity do not catabolize the isomer under usual production conditions. The results indicate that their industrial potential may be further enhanced by selection for variants that do not catabolize the sucrose substrate.     

苹果蓟属,东南亚大陆菊科一新归化属

该文报道了在泰国北部和中国西南部的菊科一个新归化属——苹果蓟属(Centratherum Cass.)。依据所采集的标本,详细描述了该属在东南亚大陆的唯一代表亚种苹果蓟 [Centratherum punctatum Cass. Subsp. fruticosum(S. Vidal)K. Kirkman ex Shih H. Chen, M. J. Wu & S. M. Li],并配以插图。苹果蓟与其所在属的其他种的区别在于该亚种叶片菱形至椭圆形,叶缘具不规则锯齿,先端微尖,总苞片基部硬化,先端无芒。文中还提供了苹果蓟的分布图,并讨论了其可能的传播途径。     

Do homomorphic and heteromorphic self-incompatibility systems have the same sporophytic mechanism?

The view put forward by some authors that flowering plant self-incompatibility mechanisms of the homomorphic sporophytic and heteromorphic sporophytic types have a close evolutionary relationship, with one form being evolved from the other, or both forms directly evolved from ancestors with homomorphic gametophytic incompatibility, is challenged. A review is provided of the various facets of each of the three main self-incompatibility systems, including a detailed summary of our current knowledge of the rejection mechanism, to demonstrate that the implicit assumption that these systems have a common S locus, and also evolutionary theories linking the systems, need to be treated with considerable caution.     

锦鸡儿属(豆科)一新组合

中间锦鸡儿(Caragana korshinskii var. intermedia), 与柠条锦鸡儿(C. korshinskii)和小叶锦鸡儿(C. microphylla)是近缘种.但以前有不同的分类学名称和处理,本文把它作为柠条锦鸡儿的一个变种(C. korshinskii Kom. var. intermedia (Kuang et H. C. Fu) M. L. Zhang & G. H. Zhu)来处理.     

Waste molasses alone displaces glucose-based medium for microalgal fermentation towards cost-saving biodiesel production

The by-product of sugar refinery—waste molasses was explored as alternative to glucose-based medium of Chlorella protothecoides in this study. Enzymatic hydrolysis is required for waste molasses suitable for algal growth. Waste molasses hydrolysate was confirmed as a sole source of full nutrients to totally replace glucose-based medium in support of rapid growth and high oil yield from algae. Under optimized conditions, the maximum algal cell density, oil content, and oil yield were respectively 70.9 g/L, 57.6%, and 40.8 g/L. The scalability of the waste molasses-fed algal system was confirmed from 0.5 L flasks to 5 L fermenters. The quality of biodiesel from waste molasses-fed algae was probably comparable to that from glucose-fed ones. Economic analysis indicated the cost of oil production from waste molasses-fed algae reduced by 50%. Significant cost reduction of algal biodiesel production through fermentation engineering based on the approach is expected.     

A revision of the Megachile subgenus Litomegachile Mitchell with an illustrated key and description of a new species (Hymenoptera,Megachilidae, Megachilini)

The species of Megachile subgenus Litomegachile are revised with a review of the species morphology, biology, and plant associations. A new species, Megachile pankus, is described and illustrated. Megachile mendica snowi Mitchell is elevated to species. Megachile var. nupta Cresson and Megachile texana var. cleomis Cockerell are synonymized with Megachile brevis and Megachile texana, respectively. An illustrated key for Litomegachile is also provided.     

Contribution to the nomenclatural knowledge of Fagus sylvatica woodlands of southern Italy

The present paper deals with some nomenclatural notes regarding two important southern Italian beech phytosociological associations such as Aquifolio-Fagetum and Asyneumato-Fagetum which were described about 30 years ago and which still represent the most widespread Fagus sylvatica communities in southern Italy. According to the ICPN, Aquifolio-Fagetum is confirmed to be a nomen illegitimum. This name must be substituted with the name Anemono-Fagetum (Gentile 1970) Brullo 1983. Moreover, Aremonio-Fagetum Hofmann 1961, a beech forest type very similar to the Aquifolio-Fagetum is a nomen invalidum. As far as the Asyneumato-Fagetum Gentile 1970 is concerned, in our opinion it represents a later syntaxonomical synonym of the Ranunculo brutii-Fagetum Bonin 1967. Geranio-Fagion Gentile 1970 and Lamio-Fagenion Gentile 1970 are typified. A new suballiance named Doronico-Fagenion (describing the beech community types of the low montane belt), deriving from the lowering of the rank of the former alliance Doronico-Fagion Ubaldi et al. ex Ubaldi 1995, is proposed.     

Allosteric activation of antithrombin is independent of charge neutralization or reversal in the heparin binding site

We investigate the hypothesis that heparin activates antithrombin (AT) by relieving electrostatic strain within helix D. Mutation of residues K125 and R129 to either Ala or Glu abrogated heparin binding, but did not activate AT towards inhibition of factors IXa or Xa. However, substitution of residues C-terminal to helix D (R132 and K133) to Ala had minimal effect on heparin affinity but resulted in appreciable activation. We conclude that charge neutralization or reversal in the heparin binding site does not drive the activating conformational change of AT, and that the role of helix D elongation is to stabilize the activated state.     

Plant cyclins: a unified nomenclature for plant A-, B- and D-type cyclins based on sequence organization

The comparative analysis of a large number of plant cyclins of the A/B family has recently revealed that plants possess two distinct B-type groups and three distinct A-type groups of cyclins [1]. Despite earlier uncertainties, this large-scale comparative analysis has allowed an unequivocal definition of plant cyclins into either A or B classes. We present here the most important results obtained in this study, and extend them to the case of plant D-type cyclins, in which three groups are identified. For each of the plant cyclin groups, consensus sequences have been established and a new, rational, plant-wide naming system is proposed in accordance with the guidelines of the Commission on Plant Gene Nomenclature. This nomenclature is based on the animal system indicating cyclin classes by an upper-case roman letter, and distinct groups within these classes by an arabic numeral suffix. The naming of plant cyclin classes is chosen to indicate homology to their closest animal class. The revised nomenclature of all described plant cyclins is presented, with their classification into groups CycA1, CycA2, CycA3, CycB1, CycB2, CycD1, CycD2 and CycD3.     

我国生物制品通用名命名原则的修订

对我国生物制品通用名命名原则进行了立题研究,旨在建立和完善我国生物制品通用名称命名原则。通过分析我国生物制品通用名现状、存在的问题及新型生物药发展趋势,研究比较了国际上具有主导性的药品通用名命名体系,并参照世界卫生组织国际非专利名称(WHO INN)命名原则,在中文语言识别的基础上,对我国生物制品通用名称命名原则提出了具体的增修订意见。     

Modification of the Oligidic Medium for Axenic Culture of Aphelenchoides rutgersi

Aphelenchoides rutgersi was axenically cultured in modified Soytone, yeast extract, lyophilized chick embryo extract medium (3% ST:2% YE:20% CEE-L, w/v:w/v:v/v). Earlier formulations used 10% CEE, v/v, before the manufacturer changed the preparation. After reestablishing A. rutgersi in medium that permitted continuous subcultivadon and reproduction, a second medium was tested that contained 0.5% sucrose and 0.5% Lipid Concentrate. The commercially available Lipid Concentrate made it possible to incorporate nonaqueous soluble chemicals into the medium. In addition, 0.1% Fast Green #3 was added to both media to visually demonstrate active ingestion of nutriment.     

无量山石豆兰, 中国兰科植物一新记录种

报道了中国兰科植物一新记录种,即无量山石豆兰(Bulbophyllum pinicola)。对其进行了特征描述并与相似种Bulbophyllum acutiflorum做了对比,并提供了彩色照片。无量山石豆兰的萼片披针形,先端渐尖,花瓣三角状披针形,先端渐尖,唇瓣稍向外下弯而易与相似种区分。     

A BROAD SPECTRUM ARTIFICIAL SEA WATER MEDIUM FOR COASTAL AND OPEN OCEAN PHYTOPLANKTON1

A new artificial seawater medium has been tested with 83 strains of coastal and open ocean phytoplankton from 11 different algal classes. The cultures were carried through four transfers, representing a period of eight weeks for most species. Only three species could not be maintained in the enriched artificial seawater, and 16 species, mainly from the Prymnesiophyceae and Dinophyceae, had reduced final cell yields compared to those grown in enriched natural seawater. Since 77% of the species tested grew equally well in enriched artificial or natural seawater and more than 95% could be maintained in the artificial medium, this recipe is useful over a broad spectrum of species. The artificial seawater base was enriched with a modified ES enrichment solution; the primary modifications were the omission of Tris and the addition of Si. Enriched medium was autoclaved without precipitation by lowering the pH before autoclaving. This was accomplished by adding equimolar amounts of Na-HCO₃ and HCl which produced NaCl and CO₂ during the heating process. When no pH buffer was used, precipitation could only be avoided by autoclaving the artificial seawater base as two separate salt solutions (with Ca and Sr separated from CO₃^?2 and SO₄^?2), cooling, mixing and aseptically adding the sterilized enrichment solution.     

Biological and biochemical modulation of Trichomonas vaginalis KT9 isolate after shifting of culture medium from TPS-1 into TYM

To evaluate the biological and biochemical characteristics of Trichomonas vaginalis KT9 isolate, the growth and size of trichomonads, pathogenicity in mouse, protein profiles and proteinase activity were examined after shifting the medium from TPS-1 into TYM. Generation time of trichomonads in TYM medium was 4.5 hr in comparison to TPS-1 with 7.1 hr. Size of trichomonads cultured in TPS-1 medium (8.5 ± 0.9 × 6.0 ± 0.9 µm) was significantly smaller than those in TYM medium (10.9 ± 1.4 × 8.2 ± 0.9 µm). Trichomonads cultured in TYM medium produced subcutaneous abscess in 9 out of 10 mice, whereas those in TPS-1 medium produced abscesses in 2 out of 10 mice. In SDS-PAGE, trichomonad lysates from both media showed ten common bands. However, trichomonads in TYM medium showed additional bands of 136 kDa, 116 kDa and 40 kDa in comparison to those in TPS-1 with 100 kDa. By immunoblot with T. vaginalis-immunized rabbit sera, T. vaginalis cultivated in both TYM and TPS-1 media showed 5 common bands, and unique bands of 116 kDa, 105 kDa, and 86 kDa were observed in trichomonads in TYM while a 140 kDa band in those in TPS-1. In gelatin SDS-PAGE, trichomonads in TYM degraded gelatin stronger than those in TPS-1. Also protease activity of trichomonads in TYM was significantly higher than that of trichomonads in TPS-1 using Bz-Pro-Phe-Arg-Nan as a substrate. According to the results, it is assumed that the shift from TPS-1 into TYM medium for cultivation of T. vaginalis might modulate the biological and biochemical properties of T. vaginalis in vitro.     

Anchorage-independent growth of breast carcinoma cells is mediated by serum exosomes

We hereby report studies that suggest a role for serum exosomes in the anchorage-independent growth (AIG) of tumor cells. In AIG assays, fetal bovine serum is one of the critical ingredients. We therefore purified exosomes from fetal bovine serum and examined their potential to promote growth of breast carcinoma cells in soft agar and Matrigel after reconstituting them into growth medium (EEM). In all the assays, viable colonies were formed only in the presence of exosomes. Some of the exosomal proteins we identified, have been documented by others and could be considered exosomal markers. Labeled purified exosomes were up-taken by the tumor cells, a process that could be competed out with excess unlabeled vesicles. Our data also suggested that once endocytosed by a cell, the exosomes could be recycled back to the conditioned medium from where they can be up-taken by other cells. We also demonstrated that low concentrations of exosomes activate MAP kinases, suggesting a mechanism by which they maintain the growth of the tumor cells in soft agar. Taken together, our data demonstrate that serum exosomes form a growth promoting platform for AIG of tumor cells and may open a new vista into cancer cell growth in vivo.     

Nomenclature of fatty acid-binding proteins

Summary A variety of designations is currently being used to refer to cellular fatty acid-binding proteins (FABPs). Besides from the use of other general names (e.g. Z protein), confusion mostly arises from the application of various abbreviations and symbols to denote the tissue(s) of origin and cellular localization (cytoplasm, plasma membrane) of a specific FABP. In order to minimize confusion a more unified and rational nomenclature is proposed, which is based on application of the formula X-FABPy. The prefix X is a capital letter indicating the tissue of greatest abundance, the suffix Y similarly denotes the (sub)cellular localization of the protein. The general and functional name fatty acid-binding protein (FABP) is preferred for the cellular proteins with the property to bind fatty acids, unless future research reveals that the binding of fatty acids is not the primary biological property or physiological role of (some of) these proteins.