Screening, Identification and Fermentation Property of a Yeast Strain R6 Accumulating Alpha-ketoglutaric Acid

A yeast strain R6 was obtained by the method of thiamine（VB1） auxotrophic negative selection from the edible oil-polluted soil in Zibo, China. Physiological and biochemical experiments revealed that strain R6 shared common feature with Rhodotorula mucilaginosa according to the API 20 C AUX yeast identification system which has been tested previously. Furthermore, the 18 S r DNA gene of strain R6 was amplified and sequenced. Phylogenetic analysis based on the 18 S r DNA sequence and the relatives indicated that R6 shared 99% homologies with the members of R. mucilaginosa, suggesting that strain R6 belonged to R. mucilaginosa.Investigation showed that strain R6 possessed the capacity of accumulating exocellular alpha-ketoglutaric acid（alpha-KG）. Finally, the fermentation conditions of R6 to accumulate alpha-KG was optimized by controlling each single fermenting variable and detected through high performance liquid chromatography（HPLC）. Results showed that both VB1 and Ca CO3 in fermentation medium were the key factors influencing the cumulant of alpha-KG. The discovery of natural auxotrophic strain R6 not only broadened the microbial resource which can achieve lots of alpha-KG production through fermentation, but also laid a foundation for further fermentation regulation to achieve excessive alpha-KG accumulation.

Taxonomic Clarification of A Well-Known Pathogenic Scuticociliate,Miamiensis avidus Thompson&Moewus,1964(Ciliophora,Scuticociliatia)

Miamiensis avidus Thompson & Moewus,1964,is a cosmopolitan and well-known marine pathogenic ciliated protist.However,the taxonomy of this species up to now has remained controversial,especially with respect to the validity of the morphologically similar species,Philasterides dicentrarchi,which was considered as a junior synonym of M.avidus.In this study,a population of M.avidus was collected from the skin of pharaoh cuttlefish(Sepia pharaonis) cultured near the East China Sea,Ningbo,China and its morphology and phylogeny were investigated in detail based on living characters,infraciliature,small subunit(SSU) r DNA and ITS1-5.8 S-ITS2 region sequences.In addition,the morphometrics of a previously reported free-living population,collected from the Bohai Sea,were rechecked and analyzed.We compared the present two isolates with all historic populations of M.avidus and P.dicentrarchi,and found that their morphological characters were either highly similar or exactly identical,indicating that they are the same morphospecies.However,the phylogenetic analyses based on SSU r DNA or ITS1-5.8 S-ITS2 region sequences revealed that most M.avidus and P.dicentrarchi populations formed one clade,and the two isolates of M.avidus from Weifang and American Type Culture Collection clustered in another clade,which indicated that there might be cryptic species in Miamiensis avidus.