Fe3+胁迫对假微型海链藻中性脂累积及DGAT 表达的影响

二酰甘油酰基转移酶(DGAT)是脂类合成途径中的关键酶,其表达水平的高低影响着脂类含量的高低。微量元素Fe3+对于微藻的生长不可或缺,且影响着微藻中油脂的累积。本实验以假微型海链藻(Thalassiosira pseudonana)为研究对象,分析了铁限制(0.000 03mmol/L)和高铁胁迫(0.3 mmol/L)2种Fe3+胁迫条件下,不同种群生长期中性脂累积及二酰甘油酰基转移酶(DGAT)的基因表达受到的影响。结果显示:铁限制条件下,微藻种群生长及其中性脂的合成受到抑制,DGAT的表达量下降;高铁胁迫条件下,高浓度Fe3+可促进中性脂合成与DGAT的表达,但抑制微藻种群生长。因此,富铁条件下更利于总脂的收集。     

现浇X形桩桩承式加筋路堤三维有限元分析

桩承式加筋路堤在高速公路软基处理中得到了广泛应用,但目前对其承载变形机制还缺乏深刻认识。结合南京长江第四大桥北接线段软基加固工程,基于数值方法对现浇X形桩桩承式加筋路堤中桩土沉降、桩土应力、桩身轴力及超静孔隙水压力的发展变化规律进行了分析。研究结果表明,受土拱效应影响,路堤填筑荷载主要由桩体承担,桩及桩间土土压力在路堤填筑过程中均逐渐增大,在软基固结过程中,桩顶土压力继续增大而桩间土土压力逐渐减小,最终趋于各自的稳定值;路堤填筑过程中桩身轴力增长较快,现浇X形桩桩身上部较大部分区段存在负摩阻力,桩身中性点位置经历了先逐渐上移、而后向下移动,最终趋于稳定的过程。     

Screening and characterization of the alkaline protease isolated from PLI-1, a strain of Brevibacillus sp. collected from Indonesia’s hot springs

A total of 69 strains of thermophilic bacteria were isolated from water, soil and sediment samples from three Indonesia’s hot spring areas (Pantai cermin, Kalianda and Banyu wedang) by using Minimal Synthetic Medium (MSM). The extreme thermophile Brevibacillus sp. PLI-1 was found to produce extracellular thermophilic alkaline protease with optimal activity at 70℃ and pH 8.0-9.0. The molecular weight of the protease was estimated to be around 56 kD by SDS-PAGE. The maximum activity of the protease was 26.54 U mL-1. The protease activity did not decrease after 30 min and still retained more than 70% of relative activity after 60 min at 70℃ and pH 8.0. The ion Mg2+ was found to promote protease activity at both low and high concentrations, whereas Cu2+ and Zn2+ could almost completely inhibit the activity. Divalent cation chelator EDTA inhibited the enzyme activity by 55.06% ± 0.27%, while the inhibition caused by PMSF, Leupeptin, Pepstain A and Benzamidine were 66.78% ± 3.25%, 52.37% ± 0.25%, 62.47% ± 2.96% and 50.99% ± 0.24%, respectively. Based on these observations, the enzyme activity was conspicuously sensitive to the serine and cysteine protease inhibitors. All these results indicated that the protease isolated from the strain PLI-1 was a thermophilic protease and had a high-temperature stability and a pH stability.     

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深海沉积物宏基因组文库中产蛋白酶克隆CAPRO2的筛选及酶学性质分析

利用选择性筛选培养基从所构建的深海沉积物宏基因组文库中筛选得到一株产蛋白酶的克隆（CAPR0002）,对其进行了酶学性质分析．结果表明该酶的最适作用温度为65cc,最适作用pH值为9．0．该蛋白酶具有较好的热稳定性,在40cC以下的温度中可长期保持稳定,在50℃中处理6h后仍能保持60％的活力,在60℃下保温30rain后仍能保持约60％的活力．Ca^2＋、Mg^2＋、sr^2＋、co^2＋对该蛋白酶有明显的促进作用,而且ca^2＋的存在可明显提高该蛋白酶的热稳定性,ca^2＋、sr^2＋、c0^2＋这3种离子均在3．0mmol／dm^3。浓度时具有最高的促进作用,当浓度高于3．0mmol／dm’时,这3种离子对酶活力的促进作用减弱．Hg^2＋、Fe^2＋、cu¨对酶有明显的抑制作用．CAPR02蛋白酶在pH值为7。5～9．5时活力较高,pH值为7。5时可保持约80％的活力,pH值为9．5时保持60％的活力,而在pH值高于9．5的条件下酶活力下降较快,pH值为10．0时活力降为约15％,表明CAPR02属于碱性蛋白酶．丝氨酸蛋白酶抑制剂PMSF、E一64和AEBSF对CAPR02蛋白酶均无抑制作用,显示该酶不属于丝氨酸蛋白酶,而EDTA对酶有明显的抑制作用,表明该酶属于金属蛋白酶．     

Cold-adaptive alkaline protease from the psychrophilic Planomicrobium sp.547: enzyme characterization and gene cloning

A psychrophilic bacterium strain 547 producing cold-adaptive alkaline protease was isolated from the deep sea sediment of Prydz Bay, Antarctica. The organism was identified as a Planomicrobium species by 16S rRNA analysis. The optimal and highest growth temperatures for strain 547 were 15℃ and 30℃, respectively. The extracellular protease was purified by ammonium sulfate precipitation and DEAE cellulose-52 chromatography. The optimal temperature and pH for the activity of the purified enzyme were 35 ℃ and pH 9.0, respectively. The enzyme retained approximately 40% of its activity after 2 h of incubation at 50℃. The enzymatic activity was inhibited by 1 mmol/L phenylmethyl sulfonylfluoride (PMSF) and hydrochloride 4-(2-aminoethyl)-benzenesulfonyl fluoride (AEBSF), indicating that it was a serine protease. The presence of Ca2+ and Mn2+ increased the activity of the enzyme. The protease gene with a size of 1 269 bp was cloned from Planomicrobium sp. 547 using primers designed based on the conserved sequences of proteases in GenBank. The Planomicrobium sp. 547 protease contained a domain belonging to the peptidase S8 family, which has a length of 309 amino acid (AA) residues. The alignment and phylogenetic analysis of the AA sequence indicated that the protease belonged to the subtilisin family.     

堆载条件下单桩负摩阻力模型试验研究

通过对粉土中混凝土单桩竖向静载荷的模型试验,分析了在桩周土堆载条件下,端承桩、摩擦端承桩在桩周土含水率变化时,桩侧摩阻力、桩端阻力的变化规律及不同土层的沉降、“中性点”位置和下拽力的变化。研究发现,摩擦端承桩“中性点”位置随桩周土含水率、堆载等级的变化而变化,在最优含水率附近,土层的下陷量较大,致桩体沉降量也大,故“中性点”的位置会有所上升;随着桩周土堆载的增大,桩周土对桩侧的下拽力也增大,桩体进一步下陷,使桩土相对位移反而减少,“中性点”有所上升。     

负摩擦作用下PCC桩沉降计算

从桩-土相互作用的机制出发,建立了均质地基中PCC桩负摩阻力传递的弹性微分方程。提出了基于有限差分法的一种桩身位移的迭代算法,计算结果表明,该计算方法收敛性好,且符合实际情况。根据中性点定义得出其位置在桩身中下部,与以往的研究结果相吻合。结果还表明,PCC桩比等截面的实心桩有较小的沉降。     

动物蛋白酶1058水解ti鱼可溶性蛋白质

研究了动物蛋白酶 1 0 58水解鱼是鱼蛋白时 ,酶量、温度、作用时间对鱼是鱼蛋白质水解率和苦味的影响。结果表明 :酶浓度的提高 ,水解时间的延长 ,均使鱼是鱼蛋白水解率明显提高。最适水解温度为 50℃ ,当温度高于 50℃时 ,水解率逐渐下降 ,蛋白酶开始失活。苦味值随水解率提高而增加。用正交试验的方法 ,经综合分析和试验得出 ,该蛋白酶水解鱼是鱼的最佳条件是酶量与底物之比为 1 .5∶ 1 0 0 0 ;温度为 50℃ ;水解时间为 4h。