Ianniello Rocco G, Zotta Teresa, Matera Attilio, Genovese Francesco, Parente Eugenio, Ricciardi Annamaria
Scuola di Scienze Agrarie, Forestali, Alimentari e Ambientali, Università degli Studi della Basilicata, Potenza, Italy.
Istituto di Scienze dell'Alimentazione-CNR, Avellino, Italy.
PLoS One. 2016 Nov 3;11(11):e0164065. doi: 10.1371/journal.pone.0164065. eCollection 2016.
Aerobic and respiratory cultivations provide benefits for some lactic acid bacteria (LAB). Growth, metabolites, enzymatic activities (lactate dehydrogenase; pyruvate and NADH oxidases, NADH peroxidase; catalase), antioxidant capability and stress tolerance of Lactobacillus casei N87 were evaluated in anaerobic, aerobic and respiratory (aerobiosis with heme and menaquinone supplementation) batch cultivations with different dissolved oxygen (DO) concentrations. The expression of pox (pyruvate oxidase) and cydABCD operon (cytochrome bd oxidase complex) was quantified by quantitative Real Time polymerase chain reaction. Respiration increased biomass production compared to anaerobiosis and unsupplemented aerobiosis, and altered the central metabolism rerouting pyruvate away from lactate accumulation. All enzymatic activities, except lactate dehydrogenase, were higher in respiratory cultures, while unsupplemented aerobiosis with 60% of DO promoted H2O2 and free radical accumulation. Respiration improved the survival to oxidative and freeze-drying stresses, while significant numbers of dead, damaged and viable but not cultivable cells were found in unsupplemented aerobic cultures (60% DO). Analysis of gene expression suggested that the activation of aerobic and respiratory pathways occurred during the exponential growth phase, and that O2 and hemin induced, respectively, the transcription of pox and cydABCD genes. Respiratory cultivation might be a natural strategy to improve functional and technological properties of L. casei.
需氧培养和呼吸培养对某些乳酸菌有益。在不同溶解氧(DO)浓度的厌氧、需氧和呼吸(添加血红素和甲萘醌的需氧环境)分批培养中,评估了干酪乳杆菌N87的生长、代谢产物、酶活性(乳酸脱氢酶;丙酮酸和NADH氧化酶、NADH过氧化物酶;过氧化氢酶)、抗氧化能力和应激耐受性。通过实时定量聚合酶链反应对pox(丙酮酸氧化酶)和cydABCD操纵子(细胞色素bd氧化酶复合物)的表达进行定量。与厌氧和未添加成分的需氧培养相比,呼吸作用增加了生物量的产生,并改变了中心代谢,使丙酮酸不再积累乳酸。除乳酸脱氢酶外,所有酶活性在呼吸培养中均较高,而溶解氧为60%的未添加成分的需氧培养促进了过氧化氢和自由基的积累。呼吸作用提高了对氧化应激和冻干应激的存活率,而在未添加成分的需氧培养(溶解氧60%)中发现了大量死亡、受损和活但不可培养的细胞。基因表达分析表明,需氧和呼吸途径的激活发生在指数生长期,并且氧气和血红素分别诱导了pox和cydABCD基因的转录。呼吸培养可能是改善干酪乳杆菌功能和技术特性的一种天然策略。