小麦叶中脯氨酸测定方法的研究

本文介绍的是一种改进的测小麦脯氨酸的比色法,用721分光仪对小麦叶中脯氨酸含量进行测定,简易快速,省工省药,适于大批量样品抗旱性筛选鉴定。     

经大豆DNA溶液处理的水稻后代种子的粗蛋白和氨基酸含量分析初报

以大豆为外源DNA供体,用浸种及幼苗期浇灌法和花粉管通道法直接将大豆总DNA导入受体水稻,经常规栽培获得水稻后代种子。采用微量凯氏定氮法和氨基酸自动分析仪进行水稻后代种子糙米的粗蛋白含量和氨基酸含量的测定。结果显示:三组经大豆DNA溶液处理获得的水稻后代种子糙米粗蛋白平均含量分别为16.42%、16.80%和19.87%,与对照组相比有明显提高,统计分析都达到极显著的差异(P < 0.01);在氨基酸含量测定中,有些材料的总氨基酸(除色氨酸以外)含量高达17.20%、16.86%和16.09%,其中赖氨酸的含量分别为0.60%、0.60%和0.57%,与对照组相比也有明显提高,统计分析差异也都达到极显著水平(P < 0.01)。本试验结果充分说明,利用大豆总DNA的导入方法有可能达到迅速有效地提高稻米蛋白质及赖氨酸含量的目的。     

以DBL法作为GQA-31型谷物品质分析仪定标方法测定稻米的赖氨酸含量

<正> GQA-31型谷物品质分析仪是一台近红外光谱仪,该分析仪原设计测定蛋白质、脂肪、水分等成分。应用该类仪器测定小麦、水稻的赖氨酸含量已有报导,但在这类仪器上用染料结合法定标测定稻米的赖氨酸含量未见有报导。近年,我们进行了以染料结合法(DBL法)作为GQA 31型谷物品质分析仪的定标方法,测定稻米赖氨酸含量的研究,取得了较满意的结果,为以后大批量筛选品种资源的赖氨酸含量和高赖氨酸育种材料的测定提供了可靠、经济、快速的检测方法。     

杜邦公司培育高赖氨酸作物

赖氨酸是人类食品和动物饲料的重要组成部分。然而,它在玉米里的含量极低,而玉米又是动物饲料的主要来源。对此,杜邦公司科学家们通过把细菌(对赖氨酸终产物反应不敏感)赖氨酸合成途径中的突变体基因导入canola和大豆,从而获得了富含高赖氨酸含量的canola和大豆种子。     

稻米赖氨酸快速测定条件的探讨

以氨基酸自动分析仪测定稻米赖氨酸含量为参照标准。在“茚三酮测定赖氨酸含量”(A法)的基础上,对该法中大米蛋白质的提取温度和时间、色温度和时间及茚三酮用量等条件进行了探讨,提出了稻米赖氨酸含量快速测(B法)。提取条件为90℃5分钟,显色条件为90℃20分钟;茚三酮试剂用1 ml。用B法对10个大米样品进行测定,赖氨酸含量与氨基酸分析仪测得的赖含量基本上一致,无显著差异(P>0.05)。     

生精细胞及精子赖氨酸含量的研究

本实验取１０只Ｗｉｓｔａｒ大鼠的睾丸和附睾,睾丸石蜡切片,附睾精子涂片后用苯胺蓝染色显示赖氨酸含量。结果是睾丸生精小管中精原细胞和精母细胞染色较深即赖氨酸含量较高,精子细胞和精子染色渐淡即赖氨酸含量降低,而附睾精子显示,在附睾头部的精子染色较深,附睾尾部的精子几乎不着色,应用显微分光光度计测定附睾精子,计算出头部的精子赖氨酸含量在１左右,尾部的精子赖氨酸含量接近于零。本实验还检测了１０例正常人及１０例不育者精子的赖氨酸,结果为正常人精子的赖氨酸含量较低,不育者精子赖氨酸含量高且畸形率也高。提示精子赖氨酸含量高是核蛋白转型异常的征象,可能是男性不育的一个重要原因。     

一种提高尸胺杆菌L-赖氨酸脱羧酶活性的培养基

某些细菌能产生特异性很强的L-赖氨酸脱羧酶,将其用于L-赖氨酸脱羧以测定L-赖氨酸含量,是行之有效的方法之一。我们用Gale等、日野哲雄和Soda等先后报道过的三种培养基培养尸胺杆菌(Bacte-     

用赖氨酸脱羧酶检压法测定发酵液中L-赖氨酸的含量

<正> 前言 L-赖氨酸是一种重要的必需氨基酸,它不仅在医药上有重要的用途,而且在食品、饲料方面都有特殊的应用价值。L-赖氨酸含量的多少,常常作为衡量食品或蛋白质饲料营养价值的一个指标。由于对赖氨酸需要量的不断增加,推动了我国赖氨酸生产的发展,用发酵法生产L-赖氨酸的工厂或车间越来越多。从事这方面工作的同志希望有一个能快速测定L-赖氨酸含量的方法。 L-赖氨酸含量的测定方法,目前比较常用的有化学法(比色法)和纸上电泳法。化学     

具有酶活力高、专一性强的L-赖氨酸脱羧酶菌种的选育

L—赖氨酸脱羧酶是尸胺杆菌产生的胞内酶。用于L—赖氨酸的含量分析。其作用原理是每个分子的L—赖氨酸在其脱羧酶的作用下放出一分子的CO_2,产生的CO_2可通过检压法测定其含量。该法操作简单、结果准确。在生产上具有重要的应用价值。     

大豆和苜蓿种子中葫芦巴碱含量的测定

用高效液相色谱法测定大豆和苜蓿种子内葫芦巴碱的含量的结果表明：以氨基键合柱为固定相,乙腈和水为流动相,可快速而比较准确的测定种子中葫芦巴碱含量。大豆和苜蓿种子中葫芦巴碱的含量高于葫芦巴植物的,可作为提取葫芦巴碱的原料。     

Hemoglobin tetramers stabilized by a single intramolecular cross-link

A specific intramolecular cross-link was introduced into bovine and human hemoglobin by reaction of the deoxyhemoglobin with the dialdehyde, bispyridoxal tetraphosphate (bisPL)P₄, followed by reduction with NaBH₄. The yield of cross-linked hemoglobin is 80% in both cases, using 1 mol of (bisPL)P₄ per mol of Hb. The crosslink is confined to the chains, where it connects the N-terminal residue (valine and methionine, respectively) to a lysine on the other chain across the central cavity. The stereochemical requirements for the reaction were probed by using a rigid analogous cross-linking reagent, as well as with a mutant Hb, which has a shorter distance between the residues to which the cross-link is attached. Introduction of the cross-link into human and bovine Hb results in a five-fold and four-fold reduction in the oxygen affinity and a decrease in the Bohr Effect by 1/3 and 1/2, respectively. Oxygenation remains cooperative, albeit with a decreased Hill coefficient. The cross-linked hemoglobins are oxidized more rapidly to the ferric form, but their resistance to heat denaturation is increased. The stability of the link between the chains and their hemes is 10 times greater in both cross-linked hemoglobins that in their native counterparts. The possible application of this chemical modification for the preparation of hemoglobin-based blood substitutes is discussed.     

Selecting Lysine-Excreting Mutants of Lactobacilli for Use in Food and Feed Enrichment

Lysine analogues are used to select for lysine-excreting mutants of Lactobacillus plantarum. The use of lactobacilli that excrete lysine for the enrichment of foods and feedstuffs by fermentation is discussed. The increase in lysine content of soybean milk by a mutant of L. bulgaricus and in silage by L. plantarum is shown.     

A general linear model for the genotypic covariance between relatives under assortative mating

A linear model for the genotypic covariance between relatives under assortative mating comprising the classical linear model and the model of selective assortative mating is proposed. The general conditions on the genetical and developmental mechanisms of quantitative characters, as well as on selection and the mating system, on which the model is based, are explicitly stated and discussed. A classification of different relationships is presented and it is shown that these conditions are sufficient to obtain the genotypic covariance between relatives only if the relationship is a combination of descendant-ancestor, full sib, Type 1 and Nth uncle-niece relationships. All the traditional relationships, i.e., those for which the covariances of the relatives have been obtained in the literature, fall into this category. These conditions also ensure that the regression of the individual's genotypic value on the genotypic value or phenotype of any of its ancestors is always linear.Paper No. 6619 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, North Carolina. This investigation was supported in part by NIH Research Grant No. GM 11546 from the National Institute of General Medical Sciences     

A colorimetric enzyme assay using Escherichia coli to determine nutritionally available lysine in biological materials

A novel bacterial method is described for determining nutritionally available lysine in protein foods with a lysine auxotroph of Escherichia coli (strain M2626). Lysine-dependent synthesis of the induced enzyme bT-galactosidase is determined by a colorimetric method. With this approach sensitivity is increased ca 100-fold and assay time decreased to ca 2 h. The improved procedure was applied to the assay of lysine present as the free amino acid or in small peptides, and after enzymic pre-digestion in vitro with a mixture of pronase and intestinal peptidases, to pure proteins and a variety of feed meals and rice cultivars. In addition, heat treatment of complex samples was shown to lower their content of available lysine, as judged by the decreased nutritional response of the E.coli strain.     

大豆DNA直接导入茄子引起变异

本文报道了利用花粉管通道技术,将大豆DNA直接导入茄子,引起茄子部分表型及蛋白质、赖氨酸含量的变异,并且这种变异可以遗传。结果表明,利用此项技术,在实现作物科间的遗传物质转移,在一定条件下是可能的。 自从周光宇先生提出了利用花粉管通道技术来直接导入外源DNA并实现了棉花抗病育种以来,这项技术被愈来愈多的科学工作者所采用,使我国农业分子育种首先进入了应用阶段。这一技术的采用也引起一些科学工作者在探索植物除种间以外的,如在属间甚至科间来实现遗传物质转移可能性的极大兴趣。 1989年我们利用花粉管通道技术进行大豆种间的DNA的导入的同时,进行了大豆DNA直接导入茄科的推广品种龙茄一号茄子中,旨在探索利用花粉管通道技术来实现超远缘物种间遗传物质转移的可行性,进而使高蛋白植物大豆的优良特性在其它作物上加以利用。     

Soybean DapA mutations encoding lysine-insensitive dihydrodipicolinate synthase

In plants, the rate-limiting step in the pathway for lysine synthesis is catalyzed by the enzyme dihydrodipicolinate synthase (DS), which is encoded by the DapA gene. We previously cloned the soybean (Glycine max cv. Century) DapA gene in Escherichia coli to express functional soybean DS protein. Like the wild-type soybean DS enzyme, the DS activity encoded by the cloned gene was extremely sensitive to feedback inhibition by micromolar concentrations of lysine. Three mutants of the soybean DapA gene were constructed using PCR: one with a single amino acid substitution at codon 104, another with a single amino acid substitution at codon 112, and a mutant containing both modifications. When expressed in E. coli, the mutant DS activities were insensitive to lysine at concentrations up to 1 mM.     

近红外光谱技术快速预测大豆氨基酸

为探索近红外光谱技术在大豆氨基酸测试中的应用,寻找一种快速的检测方法,以167份大豆[Glycine max(L.)Merr.]种子为材料,采用傅里叶变换近红外光谱技术(FT-NIRS)对经高效液相色谱法(HPLC)分析的18种氨基酸含量进行模拟。结果显示:天冬氨酸(R2CV=0.85)、谷氨酸(R2CV=0.86)、丝氨酸(R2CV=0.82)、甘氨酸(R2CV=0.89)、酪氨酸(R2CV=0.83)、苯丙氨酸(R2CV=0.78)、异亮氨酸(R2CV=0.86)和色氨酸(R2CV=0.81)及15种氨基酸总和(R2CV=0.82)可利用FT-NIRS准确预测;苏氨酸、精氨酸、丙氨酸、缬氨酸、亮氨酸和胱氨酸检测模型有一定的参考价值,可用来进行相对含量的估测;而对组氨酸、赖氨酸、脯氨酸和蛋氨酸的预测不准确。本研究进一步证明,利用FT-NIRS技术预测大豆主要氨基酸组分是稳定可行的。     

Quantification of total and bioavailable lysine in feed protein sources by a whole-cell green fluorescent protein growth-based <Emphasis Type="Italic">Escherichia coli</Emphasis> biosensor

Using a fluorescent whole-cell Escherichia coli biosensor previously developed in our laboratory, we determined total and bioavailable lysine in four feed ingredients (soybean, cottonseed, meat and bone meal, and sorghum) and three complete feeds (chick starter and finisher, and swine starter). The same feed sources were analyzed for total lysine by high performance liquid chromatography (HPLC) and bioavailable lysine by chick bioassay. No significant differences were found between bioavailable lysine estimates for soybean, cottonseed, meat and bone meal, chick starter and finisher, and swine starter obtained by the fluorescent E. coli biosensor and chick bioassay. Except for sorghum, the E. coli biosensor estimates for total lysine were highly comparable to those obtained by HPLC. Comparisons were also conducted between conventionally performed optical density-based and the newly developed fluorescence-based lysine assay. The lack of significant differences in data obtained for total and bioavailable lysine by both detection modes indicated reliance and accuracy of the fluorescent E. coli biosensor. Overall results suggest that the microbial assay based on green fluorescent protein fluorescence represents a promising alternative method for lysine quantification.     

The Use of an Escherichia coli Lys- Auxotroph to Assay Nutritionally Available Lysine in Biological Materials

A new bacterial method for determining amino acids in protein foods is described. Instead of the 'natural'microbial auxotrophs e.g. Tetrahymena, Streptococcus , and Leuconostoc , currently used for such assays, an 'artificial'mutant is used, viz. an auxotroph of Escherichia coli . Test proteins (Bovine serum albumin, legume and maize meals) were predigested with a mixture of pronase and intestinal peptidases, the efficiency and extent of proteolysis being monitored by pH stat titration. Final digests were examined by Sephadex gel filtration to ensure that all protein cleavage products were small enough to pass through the E. coli cell wall and to reach its cyto-plasmic amino acid and peptide permeases. The lysine content of the meals, as determined from the growth of an E. coli lysine auxotroph upon the digests, was found to be greater than 90° of the lysine determined chemically in acid hydrolysates. Practical and theoretical advantages of using this latter type of bacterium rather than the fastidious species are discussed. In addition, the particular value of using an intestinal bacterium like E. coli to assay nutritional availability of amino acids is considered in relation to its normal utilization of digested protein foods in vivo , and the similarities between its amino acid and peptide permeases and those of the intestine.