基于基因水平研究乳酸乳球菌的有氧呼吸代谢-现代食品科技.pdfVIP

现代食品科技 Modern Food Science and Technology 2016, Vol.32, No.9 基于基因水平研究乳酸乳球菌的有氧呼吸代谢 刘飞，李柏良，杜金城，于上富，丁秀云，徐敏，霍贵成 （东北农业大学，乳品科学教育部重点实验室，黑龙江哈尔滨 150030） 摘要：本研究旨在对能和不能有氧呼吸代谢的乳酸乳球菌呼吸链基因同源性和细胞色素氧化酶活力进行分析，为确定乳酸乳球 菌的有氧呼吸必要条件奠定基础。首先，分别测定已筛选到的菌株 KLDS4.0325 和 KLDS4.1102 在发酵和有氧呼吸代谢时的生长曲线； 其次，提取 KLDS4.1102 的基因组 DNA ，随后利用 PCR 扩增其呼吸链编码基因并进行 DNA 测序；最后，从 GenBank 数据库下载 KLDS4.0325 的呼吸链编码基因序列，利用DNAMAN 软件比较这两株菌呼吸链编码基因的同源性，并测定这两株菌的细胞色素氧化 酶活性。结果表明在有氧呼吸条件下 KLDS4.0325 的生物量和pH 都显著增大，而 KLDS4.1102 在两种条件下呈现一致的生长趋势， 呼吸链编码基因的同源性分析显示 KLDS4.1102 具备完整的呼吸链，而酶活测定结果却显示 KLDS4.1102 无细胞色素氧化酶活性，这 表明 KLDS4.1102 不能进行有氧呼吸代谢不是由于编码基因缺陷造成的，可能是某些编码基因不能正常表达，导致其细胞色素氧化酶 不能正常发挥作用。 关键词：乳酸乳球菌；血红素；有氧呼吸链；同源性;细胞色素氧化酶 文章篇号：1673-9078(2016)9-56-61 DOI: 10.13982/j.mfst.1673-9078.2016.9.009 Aerobic Respiration in Lactococcus lactis based on Gene Expression LIU Fei, LI Bai-liang, DU Jin-cheng, YU Shang-fu, DING Xiu-yun, XU Min, HUO Gui-cheng (Key Lab of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, China) Abstract: The homology of genes coding for respiratory chain and cytochrome oxidase activity of Lactococcus lactis strains that can or cannot undergo aerobic respiration were compared to provide a foundation for determining the necessary conditions for aerobic respiration in Lactococcus lactis. Firstly, growth curves of the two selected strains, KLDS4.0325 and KLDS4.1102, were determined under fermentation and aerobic respiration conditions. Secondly, the genomic DNA of strain KLDS4.1102 was extracted, and its respiratory chain-encoding genes were amplified by polymerase chain reaction (PCR), followed by DNA sequencing. Finally, the sequences of genes coding for the respiratory chain of KLDS4.0325 were downloaded from the GenBank database, the homology of genes coding for the resp