Matsuoka Hiroshi, Miura Atsuto, Hori Katsutoshi
Project Research Center for Interfacial Microbiology, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya, Japan.
J Biosci Bioeng. 2009 Apr;107(4):401-8. doi: 10.1016/j.jbiosc.2008.12.001.
Although microbial degradation of oils and fats has been developed for application in wastewater treatment, microbial degraders are not always effective in the field, for example, in grease-traps installed for the treatment of wastewater from restaurants and food industries. Wastewater in grease-traps is usually in a pH range of 5.5 to 6.5 due to hydrolysis of triacylglycerol (TAG). Because many microorganisms commercialized for use in grease-traps cannot grow at pH 6.0, we screened oil-degrading microorganisms from the environment by growing in a medium at pH 6.0 containing canola oil as the sole carbon source. We succeeded in isolating the bacterial strain Burkholderia arboris SL1B1, which secretes lipase and assimilates fatty acids, and the yeast strain Candida cylindracea SL1B2, which assimilates glycerol. The former cannot utilize glycerol as a carbon source while the latter shows only faint lipase activity that cannot support its active growth on TAG. Canola oil was degraded rapidly by a pure culture of SL1B1 at pH 6.0. However, the degradation was markedly enhanced by a mixed culture of SL1B1 and SL1B2, although lipase activity during cultivation was similar between the pure and mixed cultures. This suggests that the reversible reaction proceeds in the direction of hydrolysis of TAG due to consumption of the reaction product, glycerol, by the symbiotic yeast strain. The optimum pH and temperature of lipase secreted by B. arboris SL1B1 were 8.0 and 60 degrees C, respectively. This lipase showed highly thermal stability; the residual activity after incubation at 70 degrees C for 2 h did not decline.
尽管油脂的微生物降解技术已被开发用于废水处理,但微生物降解剂在实际应用中并非总是有效,例如,在用于处理餐馆和食品工业废水的隔油池中。由于三酰甘油(TAG)的水解作用,隔油池中的废水pH值通常在5.5至6.5之间。因为许多商业化用于隔油池的微生物在pH 6.0时无法生长,所以我们通过在以菜籽油作为唯一碳源、pH为6.0的培养基中培养,从环境中筛选了油脂降解微生物。我们成功分离出了分泌脂肪酶并能同化脂肪酸的伯克霍尔德菌属树木伯克霍尔德菌SL1B1菌株,以及能同化甘油的圆柱假丝酵母SL1B2菌株。前者不能利用甘油作为碳源,而后者仅表现出微弱的脂肪酶活性,无法支持其在TAG上的活跃生长。在pH 6.0时,SL1B1的纯培养物能迅速降解菜籽油。然而,SL1B1和SL1B2的混合培养显著增强了降解效果,尽管纯培养和混合培养过程中的脂肪酶活性相似。这表明由于共生酵母菌株消耗了反应产物甘油,TAG的水解可逆反应朝着水解方向进行。树木伯克霍尔德菌SL1B1分泌的脂肪酶的最适pH和温度分别为8.0和60℃。这种脂肪酶表现出高度的热稳定性;在70℃孵育2小时后的残余活性没有下降。