Department of Agricultural Biotechnology, Faculty of Agriculture, Kahramanmaras Sutcu Imam University, Kahramanmaras, Turkey.
Electrophoresis. 2013 Aug;34(15):2218-28. doi: 10.1002/elps.201300002.
Lactococcus lactis subsp. cremoris MG1363 genome sequence was completed and encodes two flp genes flpA and flpB. Research carried out has suggested that the flpB proteins are transcriptional regulators that respond to the environmental oxygen level. A variety of flp deletion mutant strains with single and double mutation were created. Wild-type (MG1363) and its flp(-) derivatives were compared by 2DE to identify changes in protein intensity under different aerobic/anaerobic growth conditions. In total, 416 ± 20 and 444 ± 32 protein spots were quantified from anaerobic and aerobic cells, respectively, on pH 4-7 gels. Forty-five protein spots that changed were excised from 2DE gel, digested with trypsin and identified from their MALDI-TOF MS Peptide Mass Fingerprint. A variety of proteins were affected by the flp mutations and oxygen level. Some proteins were controlled by FlpA and FlpB independently and some required both FlpA and B for regulation. The identified proteins that are regulated by the Flp proteins can be grouped by biochemical function. These groups are oxidative stress, electron transfer, sugars, cell wall, ABC transporters, arginine metabolism, and pyrimidine biosynthesis pathway.
乳球菌乳亚种 MG1363 的基因组序列已经完成，其中编码了两个 flp 基因 flpA 和 flpB。研究表明，flpB 蛋白是对环境氧水平作出响应的转录调节因子。创建了具有单突变和双突变的各种 flp 缺失突变株。通过 2-DE 将野生型（MG1363）及其 flp（-）衍生物进行比较，以确定在不同需氧/厌氧生长条件下蛋白质强度的变化。在 pH4-7 凝胶上，从厌氧和需氧细胞中分别定量了 416±20 和 444±32 个蛋白质斑点。从 2-DE 凝胶中切下 45 个发生变化的蛋白质斑点，用胰蛋白酶消化，并通过 MALDI-TOF MS 肽质量指纹图谱进行鉴定。许多蛋白质受到 flp 突变和氧水平的影响。一些蛋白质由 FlpA 和 FlpB 独立控制，而一些蛋白质则需要 FlpA 和 B 来调节。由 Flp 蛋白调节的鉴定出的蛋白质可以按生化功能进行分组。这些组是氧化应激、电子传递、糖、细胞壁、ABC 转运蛋白、精氨酸代谢和嘧啶生物合成途径。
