Functional significance of amylase polymorphism in Drosophila melanogaster. III. Ontogeny of amylase and some α-glucosidases

Changes in amylase (E.C. 3.2.1.1), maltase (E.C. 3.2.1.20), sucrase, and PNPGase activities in relation to changes in wet weight and protein content were studied during the development of larvae and adult flies from two strains of Drosophila melanogaster, homozygous for different amylase alleles. All -glucosidase activities increase exponentially during a large part of larval development, parallel to the increase in weight, and drop at the end of the third instar. Amylase activity of the Amy ¹ strain follows the same pattern. In contrast, amylase activity of the Amy ^4,6 strain continues its exponential increase longer. In the third larval instar amylase activity in the Amy ^4,6 strain becomes much higher than in the Amy ¹ strain. During the first hours of adult life amylase activity of the two strains does not differ. Then Amy ^4,6 activity starts to rise and becomes much higher (4–5 times) than Amy ¹ amylase activity, which remains approximately constant. All adult enzyme activities are much higher than in larvae. Comparison of enzyme activity of amylase and -glucosidases in larvae and adults confirms that differences in amylase activities can become important only when starch is a limiting factor in the food.The investigations were supported by the Foundation for Fundamental Biological Research (BION), which is subsidized by the Netherlands Organization for the Advancement of Pure Research (Z.W.O.).     

Regulation of amylase activity in Drosophila melanogaster: Variation in the number of enzyme molecules produced by different amylase genotypes

Purified amylases from high- and low-activity variants of Drosophila melanogaster showed identical specific activities. Immunoelectrophoresis of crude larval homogenates showed severalfold differences between strains in the amounts of cross-reacting material. Control of amylase activity is trans-acting in heterozygotes between high- and low-activity variants. These results suggest the existence of polymorphic regulatory genes affecting the production levels of amylase protein in D. melanogaster.This work was supported by Grant GM-21279 from the Institute of General Medical Science of the NIH to R. C. Lewontin and by an Operating Grant from the Natural Sciences and Engineering Research Council Canada to D. A. Hickey.     

电离辐射降低大鼠胰淀粉酶和胰蛋白酶活性机制的一些初步分析

1.丙线照射后胰淀粉酶、胰蛋白酶活性明显降低,胰酶活性降低程度和胰腺重量有关。在一定照射剂量范围内,胰酶活性和腺重量随着照射剂量的增加而下降。2.同样剂量的丙线照射后,胰腺组织匀浆上清液中的胰酶活性无变化。3.丙线照射后胰腺腺泡由M受体激动剂氨甲酰胆碱引起的淀粉酶分泌量明显减少。4.电镜观察表明,丙线照射后大鼠胰腺腺泡内合成及分泌酶原的内质网、线粒体等超微结构紊乱,这可能是电离辐射后胰酶活性降低的主要原因。     

Analysis of the mouseAmy locus in recombinant inbred mouse strains

Pancreatic and salivary amylase cDNA probes have been used to search for new DNA fragment length variation among a total of 43 inbred mouse strains. Fragment length differences found with three restriction endonucleases grouped the strains into two major classes. The segregation of these variant fragments has been analyzed among several sets of recombinant inbred strains and is presented here. Previously reported differences for strains YBR and CE have been confirmed. New segregation data for carbonic anhydrase, a locus near the proximal end of mouse chromosome 3, are presented.     

The effect of sodium fluoride on cereal seed germination and seedling growth

Abstract. Fluoride has been shown to inhibit germination and seedling establishment in barley cv. Natasha, wheat cv. Axona and rice cv. Ishikari. The concentrations of fluoride were selected so as to give a satisfactory dose-response relationship and also to relate to levels of fluoride present under natural conditions. Concentrations used were zero, 0.5, 1.0, 10 and 40 mol m⁻³ sodium fluoride. In germination studies, barley was found to be least tolerant and rice most tolerant of fluoride. The inhibition was found to be a response to fluoride itself and the observed effects were not due to pH, sodium or an unspecific effect of halide ion. Phosphate, applied at two concentration levels, was unable to overcome the inhibition. Fluoride was also shown to inhibit seedling growth. The inhibitory effects of fluoride may be accounted for by a wide range of metabolic effects including inhibition of gibberellic acid–triggered alpha amylase activity during germination, and later on to inhibition of chlorophyll synthesis in the developing leaves.     

碱性淀粉酶的发酵生产及其应用研究进展

碱性淀粉酶是诸多碱性酶中的一种,指的是其最适稳定及反应pH值在碱性范围内,在碱性环境中可以保持稳定且可高效催化降解淀粉。碱性淀粉酶在纺织、洗涤剂、医药、食品等领域具有广泛应用。利用嗜碱微生物可以生产碱性淀粉酶。以下综述了碱性淀粉酶的发酵生产及其应用的研究进展。     

A kinetic model to explain the maximum in alpha-amylase activity measurements in the presence of small carbohydrates

The effect of the presence of several small carbohydrates on the measurement of the alpha-amylase activity was determined over a broad concentration range. At low carbohydrate concentrations, a distinct maximum in the alpha-amylase activity versus concentration curves was observed in several cases. At higher concentrations, all carbohydrates show a decreasing alpha-amylase activity at increasing carbohydrate concentrations. A general kinetic model has been developed that can be used to describe and explain these phenomena. This model is based on the formation of a carbohydrate-enzyme complex that remains active. It is assumed that this complex is formed when a carbohydrate binds to alpha-amylase without blocking the catalytic site and its surrounding subsites. Furthermore, the kinetic model incorporates substrate inhibition and substrate competition. Depending on the carbohydrate type and concentration, the measured alpha-amylase activity can be 75% lower than the actual alpha-amylase activity. The model that has been developed can be used to correct for these effects in order to obtain the actual amount of active enzyme.     

驳枝对早花烟草成熟期间叶中叶绿素含量、硝酸还原酶和淀粉酶活性的影响

对早花烟草驳枝后,其成熟期间中部和上部叶中叶绿素（Ch1）含量和硝酸还原酶（NR）活性下降,下部叶则增高,成熟后期的上部叶中淀粉酶（AM）活性明显增强。经驳枝的烟草,中、上部叶烘烤后叶中淀粉、氮和烟碱含量下降,下部叶中的淀粉含量无明显变化。     

Solution structure of the plant defensin VrD1 from mung bean and its possible role in insecticidal activity against bruchids

Vigna radiata plant defensin 1 (VrD1) is the first reported plant defensin exhibiting insecticidal activity. We report herein the nuclear magnetic resonance solution structure of VrD1 and the implication on its insecticidal activity. The root-mean-square deviation values are 0.51 +/- 0.35 and 1.23 +/- 0.29 A for backbone and all heavy atoms, respectively. The VrD1 structure comprises a triple-stranded antiparallel beta-sheet, an alpha-helix, and a 3(10) helix stabilized by four disulfide bonds, forming a typical cysteine-stabilized alphabeta motif. Among plant defensins of known structure, VrD1 is the first to contain a 3(10) helix. Glu26 is highly conserved among defensins; VrD1 contains an arginine at this position, which may induce a shift in the orientation of Trp10, thereby promoting the formation of this 3(10) helix. Moreover, VrD1 inhibits Tenebrio molitor alpha-amylase. Alpha-amylase has an essential role in the digestion of plant starch in the insect gut, and expression of the common bean alpha-amylase inhibitor 1 in transgenic pea imparts complete resistance against bruchids. These results imply that VrD1 insecticidal activity has its basis in the inhibition of a polysaccharide hydrolase. Sequence and structural comparisons between two groups of plant defensins having different specificity toward insect alpha-amylase reveal that the loop between beta2 and beta3 is the probable binding site for the alpha-amylase. Computational docking experiments were used to study VrD1-alpha-amylase interactions, and these results provide information that may be used to improve the insecticidal activity of VrD1.