Prenatal diagnosis of medium-chain acyl-coenzyme A dehydrogenase deficiency

A fatal case of medium-chain acyl-coenzyme A dehydrogenase deficiency is described in a patient who presented with hypoglycaemia and a gross non-ketotic dicarboxylic aciduria. Cultured skin fibroblasts released ¹⁴CO₂ from [1–¹⁴C] octanoic acid at half the normal rate. Prenatal diagnosis was undertaken in a subsequent pregnancy in which cultured amniotic fluid cells revealed a marked reduction in octanoate oxidation indicative of an affected fetus. The pregnancy was terminated and the diagnosis was confirmed by enzyme analysis of skin fibroblasts taken from the fetus. The high residual octanoate oxidation by affected fibroblasts together with the absence of any characteristic abnormality of amniotic fluid organic acids are a potential limitation to the reliability of this type of prenatal diagnosis.     

厌氧发酵产沼气过程中脱氢酶活性检测工艺及其优化

为了优化脱氢酶活性这一指标在厌氧发酵产沼气系统中的检测方法,分别以小麦秸秆、羊粪及其混合原料(质量比1∶1)为发酵底物,通过Box-Behnken试验及响应面分析法确定并优化了沼气发酵系统中碘硝基四唑紫法脱氢酶活性的检测工艺,并对脱氢酶活性与甲烷产量进行了相关性分析.结果表明,3种底物均以乙醇作为萃取剂,以甲醛作为终止剂时显色效果最好.分别以秸秆、粪便及其混合为发酵底物时,脱氢酶检测在pH值分别为6.5、7.2、7.7,反应温度均在37℃下,碘硝基四唑紫质量分数均为0.15%,反应时间均为50 min时比色效果最好.相关性分析显示,厌氧发酵产沼气系统中脱氢酶活性与甲烷产量相关性显著.     

Prenatal diagnosis of pyruvate dehydrogenase E1α subunit deficiency

Pyruvate dehydrogenase (PDH) E1α subunit deficiency is an X-linked inborn error of metabolism affecting males and females with equal frequency. The diagnosis is usually based on determination of enzyme activity, although this may present difficulties in some females because of X-inactivation patterns favouring expression of the normal X chromosome. This is a particular problem for prenatal diagnosis using chorionic villus cells where normal enzyme assay results do not necessarily exclude the diagnosis and confirmatory X-inactivation analysis may be complicated by variable methylation of active and inactive X chromosomes. We describe prenatal diagnosis in two pregnancies in a family following diagnosis of a PDH E1α deficient male. The first prenatal diagnosis was performed by enzyme assay, but by the time of the subsequent pregnancy, the underlying mutation in the affected male had been identified and direct gene analysis was possible. This study highlights the limitations of diagnosis of PDH E1α deficiency based on measurement of the gene product and illustrates the need for mutation analysis in affected individuals.     

戊二醛对紫色非硫光合细菌脱氢酶活力的影响

探讨微生物细胞对戊二醛毒性的耐受能力及应用条件的适宜范围,可为废水处理中固定化细胞的应用提供实验基础和理论依据。以紫色非硫光合细菌（PSB）为例,直接用戊二醛作用于细胞,在不同pH值、温度、戊二醛浓度、反应时间等条件下研究戊二醛对PSB脱氢酶活力的影响。结果表明：戊二醛对PSB脱氢酶活力影响的主要限制因素为戊二醛浓度和反应时间。当戊二醛浓度为0.5～1.0％,反应时间为1～2h时,PSB脱氢酶活力回收可达83％以上;PSB细胞经戊二醛处理后,热稳定性增强,pH稳定性无明显变化;并比较出热带假丝酵母菌和PSB对戊二醛的毒性耐受能力大于大肠埃希氏杆菌和芽孢杆菌yol。     

蜡样芽孢杆菌中甲酸脱氢酶基因产氢研究

通过基因筛选,成功分离并克隆到蜡样芽胞杆菌XN12（Bacillus cereus XN12）的甲酸脱氢酶基因fdhF（formate dehydrogenase）,该基因全长2937bp,GC含量39.3%,编码978个氨基酸,与已报道的蜡样芽孢杆菌Q1的fdhF基因（GenBank No.CP000227.1）同源性达到100%.将其连接在表达载体pET32a上并融合His标签,构建了重组质粒pET32a-FDHF-His,转入大肠杆菌BL21（Escherichia coli BL21）后获得了高效表达.重组菌株经IPTG诱导后经WesternBlot分析表明,重组蛋白分子量约为108kDa.通过对重组菌株产氢性能试验表明,重组菌对提高产氢率具有一定促进作用,产氢量为每消耗1mol的葡萄糖和甲酸盐分别能产生0.73mol和0.20mol的氢气.     

可见光固化树脂固定化含黄嘌呤脱氢酶细胞的研究

含黄嘌呤脱氢酶的细胞用可见光固化树脂包埋 ,经可见光照射交联 ,制备了固定化细胞 .可见光照射 3min对细胞的存活和细胞中黄嘌呤脱氢酶的活性没有影响 .固定化细胞的黄嘌呤脱氢酶降解次黄嘌呤的最适温度为 35℃ ,最适pH为 8 0 .在pH6— 8,温度低于 40℃时稳定 .连续使用 10批 ,平均降解次黄嘌呤 99 19%,酶活力保留 94 99%.