Characterization of thermotolerant Acetobacter pasteurianus strains and their quinoprotein alcohol dehydrogenases
Characterization of thermotolerant Acetobacter pasteurianus strains and their quinoprotein alcohol dehydrogenases
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DOI：
10.1007/s00253-009-2203-5
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发表时间：
2010-01-01
影响因子：
5
通讯作者：
Matsushita, Kazunobu
中科院分区：
文献类型：
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作者：
Kanchanarach, Watchara;Theeragool, Gunjana;Matsushita, Kazunobu
本实验分离到几种耐热醋酸杆菌，其中MSU 10菌株在41 ℃的琼脂平板上生长良好，在39 ℃时能耐受1.5%的乙酸或4%的乙醇，与A. pasteurianus SKU 1108以前分离。MSU 10菌株在37 A ℃下在含6%乙醇的培养基中表现出比SKU 1108菌株更高的乙酸生产率，而SKU 1108菌株在相同温度下在补充有4-5%乙醇的培养基中可以积累更多的乙酸。在37 A ℃下，耐高温菌株的发酵能力上级优于中温菌株。巴氏杆菌IFO 3191菌株在37 ℃下生长弱，即使在4%乙醇下乙酸产生也非常延迟。从MSU 10、SKU 1108和IFO 3191菌株中分离纯化了乙醇脱氢酶（ADH），并比较了它们与耐热性相关的性质。耐热菌株ADH的最适温度和热稳定性略高于中温菌株IFO 3191。更重要的是，在高温下，来自MSU 10和SKU 1108菌株的ADH比IFO 3191酶表现出更高的乙醇和乙酸抗性。此外，本研究还克隆了ADH基因，并比较了ADH亚基I、亚基II和亚基III的氨基酸序列。MSU 10或SKU 1108 ADH与IFO 3191 ADH在氨基酸残基上存在差异，可能与耐热性有关。
We isolated several thermotolerant Acetobacter species of which MSU10 strain, identified as Acetobacter pasteurianus, could grow well on agar plates at 41A degrees C, tolerate to 1.5% acetic acid or 4% ethanol at 39A degrees C, similarly seen with A. pasteurianus SKU1108 previously isolated. The MSU10 strain showed higher acetic acid productivity in a medium containing 6% ethanol at 37A degrees C than SKU1108 while SKU1108 strain could accumulate more acetic acid in a medium supplemented with 4-5% ethanol at the same temperature. The fermentation ability at 37A degrees C of these thermotolerant strains was superior to that of mesophilic A. pasteurianus IFO3191 strain having weak growth and very delayed acetic acid production at 37A degrees C even at 4% ethanol. Alcohol dehydrogenases (ADHs) were purified from MSU10, SKU1108, and IFO3191 strains, and their properties were compared related to the thermotolerance. ADH of the thermotolerant strains had a little higher optimal temperature and heat stability than that of mesophilic IFO3191. More critically, ADHs from MSU10 and SKU1108 strains exhibited a higher resistance to ethanol and acetic acid than IFO3191 enzyme at elevated temperature. Furthermore, in this study, the ADH genes were cloned, and the amino acid sequences of ADH subunit I, subunit II, and subunit III were compared. The difference in the amino acid residues could be seen, seemingly related to the thermotolerance, between MSU10 or SKU1108 ADH and IFO 3191 ADH.