猪血红蛋白的酶解及氨基酸含量的研究

猪血是一种营养价值高而价格低廉的蛋白质资源.猪血中含有丰富的蛋白质,其中血红蛋白占主要部分.因此,对血红蛋白及其水解产物的研究受到了国内外众多学者的关注.为选出适合于水解猪血红蛋白的蛋白酶,笔者采用碱性蛋白酶、木瓜蛋白酶、中性蛋白酶对猪血进行单酶和复合酶水解试验,确定碱性蛋白酶为水解猪血的适用酶.通过质谱仪测定,碱性蛋白酶酶解产物分子量集中在800～2400道尔顿,而800～1260道尔顿区间占主要部分;经全自动氨基酸分析仪测出其产物含有17种氨基酸,其中含8种必需氨基酸,必需氨基酸占总氨基酸量的50.93%,说明碱性蛋白酶酶解产物是制作食品和饲料添加剂的理想资源.     

在我国裕固族中首次发现一例Hb沈阳

Hb沈阳是1982年在我国汉族中发现的一种新的不稳定血红蛋白。1986年作者在“丝绸之路”地 区进行血红蛋白群体普查时，发现一例裕固族泳速相当于J组的异常Hb，经一级结构分析表明为。链 第26位丙氨酸被谷氨酸取代，证实此例变异体为Hb沈阳〔a,,(B7)Ala-aGlu」在裕周族中系首次报 告。根据现有Hb沈阳的分布状况及有关资料，作者分析这种异常Hb基因可能源于我国。     

透明颤菌血红蛋白的研究进展及其在发酵工业中的应用

透明颤菌血红蛋白（Vitreoscilla Hemoglobin, VHb）是20世纪70年代后期发现的一种血红蛋白，该蛋白质能使透明颤菌在低氧的环境下生存，并保持较高的生长速率。随着其作用机理的深入研究，VHb在生物工程领域的应用越来越广泛。本文总结了近年来VHb的研究进展，包括蛋白结构、细胞定位、基因改组、作用机理等方面的研究，并概述了其在微生物发酵工业中的应用。     

The biochemical basis of antimicrobial responses in Manduca sexta

Innate immunity is essential for the wellbeing of vertebrates and invertebrates. Key components of this defense system include pattern recognition receptors that bind to infectious agents, extra-and intra-cellular proteins that relay signals, as well as molecules and cells that eliminate pathogens. We have been studying the defense mechanisms in a biochemical model insect, Manduca sexta. In this insect, hemolin, peptidoglycan recognition proteins, β-1,3-glucan recognition proteins and C-type lectins detect microbial surface molecules and induce immune responses such as phagocytosis, nodulation, encapsulation, melanization and production of antimicrobial peptides. Some of these responses are mediated by extracellular serine proteinase pathways. The proteolytic activation of prophenoloxidase （proPO） yields active phenoloxidase （PO） which catalyzes the formation of quinones and melanin for wound healing and microbe killing. M. sexta hemolymph proteinase 14 （HP 14） precursor interacts with peptidoglycan or β-1,3-glucan, autoactivates, and leads to the activation of other HPs including HP21 and proPO-activating proteinases （PAPs）. PAP-1, -2 and -3 cut proPO to generate active PO in the presence of two serine proteinase homologs. Inhibition of the proteinases by serpins and association of the proteinase homologs with bacteria ensure a localized defense reaction. M. sexta HP1, HP6, HP8, HP17 and other proteinases may also participate in proPO activation or processing of spatzle and plasmatocyte spreading peptide.     

慢病毒载体介导的转人ahsp基因的β654地中海贫血小鼠的制备

目的：经慢病毒载体介导,制备转人α血红蛋白稳定蛋白基因（ahsp）的β654地中海贫血小鼠。方法：用巢式PCR从人血DNA中获得人ahsp基因,构建含有人ahsp基因的慢病毒载体,制备假病毒,通过卵周隙显微注射手段将其导入β654地贫小鼠的受精卵,经移植至假孕母鼠输卵管,最终孕育出转人ahsp基因的β654地贫小鼠;分析小鼠体内外源ahsp基因的表达情况及其遗传稳定性。结果：共获得了8只人ahsp阳性小鼠,转基因阳性率为32%（8/25）,其中3只同时具有β654突变基因;人ahsp基因在小鼠体内的表达水平维持在小鼠自身ahsp表达量的1%左右,且可稳定遗传至子代。结论：制备了转人ahsp基因小鼠,并可遗传至子代,为在个体水平上研究α血红蛋白稳定蛋白与β地贫之间的关系提供了工具。     

以聚乙二醇为层析伴侣同时制备SOD、过氧化氢酶和血红蛋白

发展了一条从红细胞裂解液中同时制备超氧化物歧化酶(SOD)、过氧化氢酶和血红蛋白的新工艺。采用0 75 %的聚乙二醇600作为层析伴侣,使血红蛋白直接流过阴离子交换层析柱,同时吸附SOD和过氧化氢酶。经过梯度洗脱获得SOD和过氧化氢酶组分,再经过疏水性相互作用层析与凝胶过滤层析相串联,使SOD和过氧化氢酶得到纯化。纯化后的SOD和过氧化氢酶的比活力分别达到15932u/mg和65918u/mg ,血红蛋白的纯度达到99.9%以上。总回收率为:SOD ,47.4% ;过氧化氢酶,29.6% ;血红蛋白,88.7%。     

藏鸡胚胎的低氧适应特征及其血红蛋白突变效应

藏鸡能在4200 m以上海拔正常繁衍生息, 但低地鸡在模拟4200 m海拔(12.0%O₂)条件下孵化却只有3.0%的孵化率. 在两品种全期低氧和阶段低氧模拟孵化实验中, 4200 m全期低氧揭示4~11天是隐性白与藏鸡阶段存活率相差最悬殊的阶段, 12~21天相差极显著, 1~3天相差不显著. 但4200 m阶段低氧只能造成两品种孵化率显著差异, 而并不足以造成低氧阶段间阶段死亡率显著差异, 结果显示藏鸡具有优越的克服低氧损害后期不良影响的能力. 对藏鸡和隐性白鸡血红蛋白7条链全部编码序列进行多态分析, 筛选到α^D链编码基因的一个低氧功能突变位点(Met- 2D(B13)-Leu). 该突变基因为藏鸡独有, 其频率随着海拔升高而升高, 而且该突变基因频率越高的群体, 孵化率越高. 以鸡的HbD晶体为模板, 对突变血红蛋白结构和功能进行了同源预测, 结果表明Leu替代Met提供了一个更疏水环境, 有利于血红素的稳定, 从而提高了血红蛋白的血氧亲和力. 突变(Met-32D(B13)-Leu)发生的时间和地点与藏鸡家养化时间和地点密切相关.     

A genetic adaptive pattern＊low hemoglobin concentration in the himalayan highlanders

Mean hemoglobin （Hb） concentration of about 3 500 subjects derived from 17 studies of Himalayan highlanders （Tibetans, Sherpas, and Ladakhis） was compared with lowlanders （Chinese Han, Indian Tamils） lived in the Himalayas, and European climbers during Everest expeditions as well as Andean natives. The results found that Hb concentration in Himalayan highlanders was systemically lower than those reported for Andean natives and lowland immigrants. These comparative data demonstrated that a healthy native population may successfully reside at high altitude without a significant elevation in Hb, and the lower Hb levels of Himalayan highlanders than those of migrated lowlanders and Andean natives are an example of favourable adaptation over the generations. In addition, excessive polycythemia has frequently been used as a marker of chronic mountain sickness （CMS）. Altitude populations who have a higher Hb concentration also have a higher incidence of CMS. The low Hb in Himalayans suggested as showing adaptation over many generations in Tibetan stock. Recent work in Tibet, suggested that Tibetans there may have adapted to high altitude as a result of evolutionary pressure selecting for genes which give an advantage at altitude. All of the population genomic and statistical analysis indicated that EPAS1 and EGLN1 are mostly likely responsible for high altitude adaptation and closely related to low Hb concentration in Tibetans. These data supported the hypothesis that Himalayan highlanders have evolved a genetically different erythropoietic response to chronic hypoxia by virtue of their much longer exposure to high altitude.