Lei Lei, Alcolombri Uria, Tawfik Dan S
Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel.
Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel.
Methods Enzymol. 2018;605:269-289. doi: 10.1016/bs.mie.2018.03.004. Epub 2018 Apr 27.
Dimethyl sulfide (DMS) is released at rates of >10 tons annually and plays a key role in the oceanic sulfur cycle and ecology. Marine bacteria, algae, and possibly other organisms release DMS via cleavage of dimethylsulfoniopropionate (DMSP). DMSP lyases have been identified in various organisms, including bacteria, coral, and algae, thus comprising a range of gene families putatively assigned as DMSP lyases. Metagenomics may therefore provide insight regarding the presence of DMSP lyases in various marine environments, thereby promoting a better understanding of global DMS emission. However, gene counts, and even mRNA levels, do not necessarily reflect the level of DMSP cleavage activity in a given environmental sample, especially because some of the families assigned as DMSP lyases may merely exhibit promiscuous lyase activity. Here, we describe a range of biochemical profiling methods that can assign an observed DMSP lysis activity to a specific gene family. These methods include selective inhibitors and DMSP substrate analogues. Combined with genomics and metagenomics, biochemical profiling may enable a more reliable identification of the origins of DMS release in specific organisms and in crude environmental samples.
二甲基硫醚(DMS)每年的释放量超过10吨,在海洋硫循环和生态中起着关键作用。海洋细菌、藻类以及可能的其他生物通过二甲基巯基丙酸内盐(DMSP)的裂解来释放DMS。在包括细菌、珊瑚和藻类在内的各种生物中已鉴定出DMSP裂解酶,因此构成了一系列被假定为DMSP裂解酶的基因家族。因此,宏基因组学可能有助于深入了解各种海洋环境中DMSP裂解酶的存在情况,从而促进对全球DMS排放的更好理解。然而,基因计数,甚至mRNA水平,不一定能反映给定环境样品中DMSP裂解活性的水平,特别是因为一些被指定为DMSP裂解酶的家族可能仅仅表现出混杂的裂解酶活性。在这里,我们描述了一系列生化分析方法,这些方法可以将观察到的DMSP裂解活性归属于特定的基因家族。这些方法包括选择性抑制剂和DMSP底物类似物。结合基因组学和宏基因组学,生化分析可能能够更可靠地确定特定生物和原始环境样品中DMS释放的来源。