Bioengineered. 2013 Mar-Apr;4(2):72-7. doi: 10.4161/bioe.22461. Epub 2012 Oct 23.
Harmful algal blooms caused by phytoplankton can occur in all aquatic environments. Some of the algae present in these blooms are capable of producing extremely potent toxins. Due to climate change and eutrophication, harmful algal blooms are increasing on a global scale. One kind of toxin producing algae are those that produce okadaic acid, its derivatives (dinophysistoxin-1 and 2), and microcystins. These toxins are potent inhibitors of protein phosphatase 2A, so this protein is used to detect the mentioned toxins in natural samples. Originally protein phosphatase 2A purified from animal tissues was used, but enzyme activity and stability fluctuations prevented the use of the enzyme in detection kits. Expression of the enzyme as a recombinant protein provided a solution to this problem. For this purpose, several strategies have been followed. We evaluated the activity, specificity and stability of the human protein phosphatase 2A catalytic subunit α expressed in insect larvae and showed that this expression system can be a reliable source of high quantities of stable enzyme.
赤潮是由浮游植物引起的,可发生于所有水生环境中。这些赤潮中的一些藻类能够产生极其强效的毒素。由于气候变化和富营养化,有害藻类在全球范围内呈上升趋势。产生冈田酸、其衍生物(短裸甲藻毒素-1 和 2)和微囊藻毒素的藻类是产生毒素的一种。这些毒素是蛋白磷酸酶 2A 的强效抑制剂,因此该蛋白被用于检测天然样品中的所述毒素。最初使用的是从动物组织中纯化的蛋白磷酸酶 2A,但酶活性和稳定性波动使该酶无法用于检测试剂盒中。作为重组蛋白表达该酶提供了一种解决方案。为此,我们采用了几种策略。我们评估了在昆虫幼虫中表达的人蛋白磷酸酶 2A 催化亚基 α 的活性、特异性和稳定性,并表明该表达系统可以作为大量稳定酶的可靠来源。