球孢白僵菌胞外蛋白酶及类枯草杆菌蛋白酶的诱导

研究了接种物和诱导物对球孢白僵菌总胞外蛋白酶及类枯草杆菌蛋白酶（Ｐｒ１）产生的影响。结果表明,球孢白僵菌不同接种物产生的总胞外蛋白酶及Ｐｒ１活性有明显差异;不同诱导物对产酶水平也有较大影响。其中芽管状物在蝉蜕诱导液中的总胞外蛋白酶和Ｐｒ１活性均最高,诱导１２ｈ酶活性达高峰,不同来源的菌丝产酶水平有差异,在诱导液中,接种物总胞外蛋白酶和Ｐｒ１的变化趋势一致,但总胞外蛋白酶活性和Ｐｒ１活力间没有直接的     

生物托盘土壤微生物及酶活性的分布状况

利用稀释法和比色法对生物托盘根际和非根际土壤微生物数量和酶活性进行测定。结果表明:土壤微生物和酶在植物根际有着明显的根际效应,各级托盘中微生物在根际的数量均大于相应微生物在非根际的数量;在各级托盘中均存在细菌>放线菌>真菌的数量关系。而对土壤蛋白酶和脲酶活性的研究发现根际酶活性明显高于非根际酶活性。     

微曝气条件下S-TE剩余污泥溶解性研究

研究了微曝气、不同温度条件下S-TE预处理对剩余污泥溶解和各种化学组分变化的影响.结果表明,S-TE污泥溶解存在2种反应(酶催化反应和热解反应)和2个过程:嗜热菌胞外酶(主要为蛋白酶和淀粉酶)首先解聚污泥胶团,进而溶解细菌的细胞壁,水解胞内有机物质.接种嗜热菌Bacillus stearothermophilus sp.AT06-1比不接种促进了污泥悬浮固体的溶解,接种条件下最适宜的溶解温度为65℃,此温度下,污泥VSS和TSS溶解率相对于不接种试验提高程度最大,2 d时VSS、TSS溶解率分别达到34.09%和24.16%,比不接种试验同期分别提高了7.57%和6.87%;微曝气条件下SCOD和VFA得到累积,最大累积量达到4 531 mg/L和2 319 mg/L,有利于厌氧消化;此时蛋白酶活性提高也最大.污泥溶解产生的蛋白质被蛋白酶水解,蛋白质浓度先升高后降低.     

鳗弧菌(Vibrio anguillarum)胞外产物中蛋白酶的纯化及其性质

对鳗弧菌 (Vibrioanguillarum)胞外产物中蛋白酶的纯化方法及其性质进行了研究 .其中 ,蛋白酶的纯化步骤包括 :(1)硫酸铵沉淀 ;(2 )SephadexG 10 0凝胶过滤 ;(3)DEAE Sepharose色谱分离 ;(4)DEAE Cellulose (DE 32 )色谱分离 .经上述纯化步骤得到两种蛋白酶 ,经SDS PAGE分析 ,Mr分别为 37.4× 10 3 和 33.1× 10 3 .以偶氮酪蛋白 (azocasein)作底物测其水解酶活性 ,结果表明二者差别显著 ,前者明显高于后者 .而且 ,Mr37.4× 10 3 的蛋白酶在pH 7～ 10范围内均显示较高活性 ,在 4 0～ 6 0℃范围内稳定性好 .结果表明该蛋白酶是一种金属蛋白酶 .关于该蛋白酶对牙鲆(Paralichtysolivaceus)的毒性作用尚在研究之中 .图 6表 3参 17     

Prenatal diagnosis of a lethal form of Netherton syndrome by SPINK5 mutation analysis

Netherton syndrome (NS) is a severe autosomal recessive ichthyosis with no specific treatment or prenatal diagnosis available at present. The recent identification of SPINK5, which encodes a serine protease inhibitor, as the defective gene enables DNA-based prenatal diagnosis to be carried out. Here we report the first direct molecular prenatal diagnosis of a lethal form due to a recurrent SPINK5 mutation in three consanguineous Turkish families. XmnI restriction enzyme digestion and DNA sequencing demonstrated that each deceased affected child was homozygous for mutation 153delT inherited from each parent. Analysis of fetal DNA from amniotic fluid cells in Family 1 and from a chorionic villus sampling in Family 3 showed that the fetus was heterozygous for 153delT in both cases. The pregnancies were carried to term and the newborns were unaffected. In Family 2, fetal DNA analysis from chorionic villus biopsy showed in a first pregnancy that the fetus was homozygous for 153delT. The pregnancy was terminated at 13 weeks and DNA analysis of fetal keratinocytes confirmed the prenatal prediction. In a second pregnancy in Family 2, fetal DNA analysis showed heterozygosity for 153delT, and the pregnancy was continued. Direct SPINK5 mutation analysis in families at risk for NS represents the first early, rapid and reliable method for prenatal diagnosis of this life-threatening form of ichthyosis. Copyright © 2002 John Wiley & Sons, Ltd.     

地衣芽孢杆菌2709和 6816碱性蛋白酶基因在大肠杆菌中的克隆、表达及序列分析

用PCR方法从地衣芽孢杆菌 2 70 9和 6 816中扩增了碱性蛋白酶基因 (apr1和apr2 ) ,扩增的DNA片段插入到大肠杆菌载体 pET -2 8a中 ,构建成重组分泌型表达载体pAPR1、pAPR2 .pAPR1、pAPR2中碱性蛋白酶基因在大肠杆菌宿主JM 10 9(DE3)中得到表达 .SDS -PAGE分析显示融合表达产物的分子量均为 30 .5× 10 3 ,同核酸序列测定所推导的值相符 .表达产物分别占细胞总蛋白的 8.0 %和 7.5 % .2 70 9重组菌所得酶活为 12 10u/mL ,6 816重组菌所得酶活为 1175u/mL .研究发现 ,重组的碱性蛋白酶在进入大肠杆菌周质空间时可能存在前肽自动脱落的现象 .同时 ,对地衣芽孢杆菌 2 70 9碱性蛋白酶基因序列进行了测定和比较分析 ,发现与地衣芽孢杆菌 6 816碱性蛋白酶基因的同源性为 98% .图 5参 11     

Degradation of Polylactide by Commercial Proteases

Fifty-six commercially available proteases were tested for polylactide-degrading activity. Little or no activity was found in acid and neutral proteases, while some alkaline proteases formed appreciable amounts of lactic acid from polylactide. These polylactide-degrading proteases were derived from Bacillus species and had catalytic activity even under neutral, as well as alkaline, conditions. Savinase (Novo Nordisk) degraded polylactide the fastest among the enzymes tested and its specific activity corresponded to about one-half of proteinase K. Polylactide-degrading activity was not always present in the enzymes that affected keratin, while polylactide-degrading proteases commonly hydrolyzed keratin. A significant correlation was observed between degrading activities of polylactide and keratin in alkaline proteases.     

Effects of metolachlor on activities of enzymes in a malaysian soil

Abstract

Effects of metolachlor on the activities of cellulase, phosphatase, urease and protease in a loamy sand soil, were evaluated for up to 28 days. Metolachlor at 10 ppm and above caused a reduction in phosphatase and urease activities for the entire period of study. The cellulase activity decreased during the very first day of incubation compared to that of the control but a recovery trend was obvious at all concentrations from day 14 onwards. Protease activity was also reduced in the metolachlor treated soil and the lower activity was observed during the entire study.     

不同存放时间太湖蓝藻产沼气潜力

对不同存放时间蓝藻(新鲜蓝藻、存放3个月、6个月和9个月)进行厌氧消化处置,分析反应过程中沼气量、代谢产物和酶活等变化,获得各自的产沼气潜力。研究结果表明,存放6个月蓝藻组的产沼气率最大,为120.5 mL/g TS,是新鲜蓝藻组的1.29倍。Gompertz模型拟合表明,随着存放时间的延长,最大产沼气量(Pmax)先增大后减小,最大比产气率一直增加。存放3个月蓝藻组的VFA在第6天达到最大,为1 575.4 mg/L,而存放9个月蓝藻组的VFA在第2天达到最大,为1 116.5 mg/L。存放3个月蓝藻组的最大中性蛋白酶酶活为1 203.4μg/(g TS.h),比新鲜蓝藻组提高212.8μg/(gTS.h)。脱氢酶酶活随着蓝藻存放时间的增加呈现先增加后减小的趋势。