Saliola Michele, Bartoccioni Paola Chiara, De Maria Ilaria, Lodi Tiziana, Falcone Claudio
Dipartimento di Biologia Cellulare e dello Sviluppo, Università di Roma "La Sapienza" Rome, Italy.
Eukaryot Cell. 2004 Jun;3(3):589-97. doi: 10.1128/EC.3.3.589-597.2004.
We have isolated a Kluyveromyces lactis mutant unable to grow on all respiratory carbon sources with the exception of lactate. Functional complementation of this mutant led to the isolation of KlSDH1, the gene encoding the flavoprotein subunit of the succinate dehydrogenase (SDH) complex, which is essential for the aerobic utilization of carbon sources. Despite the high sequence conservation of the SDH genes in Saccharomyces cerevisiae and K. lactis, they do not have the same relevance in the metabolism of the two yeasts. In fact, unlike SDH1, KlSDH1 was highly expressed under both fermentative and nonfermentative conditions. In addition to this, but in contrast with S. cerevisiae, K. lactis strains lacking KlSDH1 were still able to grow in the presence of lactate. In these mutants, oxygen consumption was one-eighth that of the wild type in the presence of lactate and was normal with glucose and ethanol, indicating that the respiratory chain was fully functional. Northern analysis suggested that alternative pathway(s), which involves pyruvate decarboxylase and the glyoxylate cycle, could overcome the absence of SDH and allow (i) lactate utilization and (ii) the accumulation of succinate instead of ethanol during growth on glucose.
我们分离出了一株乳酸克鲁维酵母突变体,该突变体除乳酸外无法在所有呼吸性碳源上生长。对该突变体进行功能互补,从而分离出了KlSDH1基因,该基因编码琥珀酸脱氢酶(SDH)复合体的黄素蛋白亚基,这对于需氧利用碳源至关重要。尽管酿酒酵母和乳酸克鲁维酵母中SDH基因的序列保守性很高,但它们在这两种酵母的代谢中具有不同的相关性。事实上,与SDH1不同,KlSDH1在发酵和非发酵条件下均高表达。除此之外,与酿酒酵母相反,缺乏KlSDH1的乳酸克鲁维酵母菌株在有乳酸存在的情况下仍能生长。在这些突变体中,在有乳酸存在的情况下,氧气消耗量是野生型的八分之一,而在有葡萄糖和乙醇存在时氧气消耗量正常,这表明呼吸链功能完全正常。Northern分析表明,涉及丙酮酸脱羧酶和乙醛酸循环的替代途径可以克服SDH缺失的影响,并允许(i)利用乳酸以及(ii)在以葡萄糖为碳源生长期间积累琥珀酸而不是乙醇。