定点突变提高过氧化氢酶热稳定性和催化效率

过氧化氢酶催化过氧化氢分解形成水和氧,主要作用是保护细胞免受活性氧(ROS)的损伤,尽管其在工业上被广泛使用,但因热稳定的不足限制了它的使用前景。本研究以Bacillus pumilus ML413来源的血红素过氧化氢酶(KatX2)为研究对象,采用定点突变技术提高其热稳定性及催化效率。利用PoPMuSiC algorithm软件计算并预测了其潜在突变位点的折叠自由能,根据预测的结果,选择了其中自由能最低的3个位点(K117V,K117I和K117M)进行突变。结果发现,对K117位点进行突变,可以有效提高KatX2的热稳定性,突变体K117V在60℃下半衰期比野生型KatX2提高38 min,催化效率较突变前提高31.7%。根据结构分析发现,该点突变并未改变KatX2的二级结构。突变体K117V具有良好的工业应用潜力。

通过改造底物结合区氨基酸疏水性提高氨肽酶热稳定性

通过提高枯草芽孢杆菌来源的氨肽酶底物结合区域氨基酸的疏水性,提高氨肽酶YwaD的热稳定性。基于氨肽酶底物结合区域氨基酸特性分析,选定亲水性氨基酸残基N385为目标,通过定点突变使其突变为疏水氨基酸残基L385,获得突变体N385L。结果表明,与野生型ywaD相比,突变体N385L酶热稳定性明显提高,在80℃放置30 min,野生型YwaD完全失活,而突变体N385L仍保留20%的酶活力。另外,突变型N385L对底物氨基酰基-硝基苯胺的亲和力提高了47.4%,催化效率提高了28.5%。该研究有利于提高氨基肽酶工业应用前景。

Evaluating Anaerobic Digestion Technology in Reducing the Quantity of Solid Waste: Case of Kigali Dumpsite

Biodegradable organic matter constitutes a great portion of Municipal solid waste and comprises organic material which can be broken down by bacteria like paper, card, green waste, food waste, miscellaneous items with an organic element and fine materials. This paper mainly evaluated the potential of anaerobic digestion technology in reducing the quantity of solid waste destined to the dumpsite in Kigali city. The paper evaluates the viability of using biodegradable waste meant to the land where the biogas is produced and undertaking its cost & benefit analysis for chemical, physical and biological characteristics of municipal solids waste on anaerobic digestion technology. The quantity of municipal waste generated in Kigali city was used for designing the biodigester required. The paper indicates that the organic waste in Kigali city produces 457 L/kg DM of methane and the overall assessed value of methane was 51,384,375 L with the electricity derived from the methane of 180,873 KWh which is 54% of the daily demand in Kigali city. The volume of the biodigester was found to be 58,065 m3. Based on the energy recovered, revealed that cities will benefit this research for the population demand for the increased electricity.