Yasuda Chika, Iijima Hiroko, Sukigara Haruna, Osanai Takashi
School of Agriculture, Meiji University, Kanagawa, Japan.
Bio Protoc. 2017 May 5;7(9):e2257. doi: 10.21769/BioProtoc.2257.
Succinate and lactate are commodity chemicals used for producing bioplastics. Recently, it was found that such organic acids are excreted from cells of the unicellular cyanobacterium sp. PCC 6803 under dark, anaerobic conditions. To conduct the dark, anaerobic incubation, cells were concentrated within a vial that was then sealed with a butyl rubber cap, following which N gas was introduced into the vial. The organic acids produced were quantified by high-performance liquid chromatography via post-labeling with bromothymol blue as a pH indicator. After separation by ion-exclusion chromatography, the organic acids were identified by comparing their retention time with that of standard solutions. These procedures allow researchers to quantify the organic acids produced by microorganisms, contributing to knowledge about the biology and biotechnology of cyanobacteria.
琥珀酸和乳酸是用于生产生物塑料的商品化学品。最近，人们发现这些有机酸在黑暗、厌氧条件下从单细胞蓝藻PCC 6803的细胞中排出。为了进行黑暗、厌氧培养，将细胞浓缩在一个小瓶中，然后用丁基橡胶帽密封，接着将氮气引入小瓶。通过以溴百里酚蓝作为pH指示剂进行后标记，利用高效液相色谱法定量所产生的有机酸。通过离子排斥色谱分离后，通过将它们的保留时间与标准溶液的保留时间进行比较来鉴定有机酸。这些程序使研究人员能够定量微生物产生的有机酸，有助于了解蓝藻的生物学和生物技术。
