Permeabilization of Microbacterium oxylans shifts the conversion of puerarin from puerarin-7-O-glucoside to puerarin-7-O-fructoside.
The main product of the conversion of puerarin by unpermeabilized cells of bacterium Microbacterium oxydans CGMCC 1788 was puerarin-7-O-glucoside (241 +/- 31.9 microM). Permeabilization with 40% ethanol could not increase conversion yield, whereas it resulted in change of main product; a previous trace product became a main product (213 +/- 48.0 microM) which was identified as a novel puerarin-7-O-fructoside by electrospray ionization time-of-flight MS, (13)C NMR, (1)H NMR, and GC-MS analysis of sugar composition, and puerarin-7-O-glucoside became a trace product (14.8 +/- 5.4 microM). However, the extract from cells of M. oxydans CGMCC 1788 permeabilized with ethanol converted puerarin to form 113.9 +/- 27.7 microM puerarin-7-O-glucoside and 187.8 +/- 29.5 microM puerarin-7-O-fructoside under the same conditions. When unpermeabilized intact cells were recovered and used repeatedly for the conversion of puerarin, with increase of reuse times, the yield of puerarin-7-O-glucoside gradually decreased, whereas the yield of puerarin-7-O-fructoside increased gradually in the conversion mixture. The main product of the conversion of puerarin by the tenth recycled unpremerbilized cells was puerarin-7-O-fructoside (288.4 +/- 24.0 microM). Therefore, the change of permeability of cell membrane of bacterium M. oxydans CGMCC 1788 contributed to the change of conversion of the product's composition.
Yu
Cigang
C
Jiangsu Engineering and Technology Research Center for Industrialization of Microbial Resources, Jiangsu Key Laboratory for Biodiversity and Biotechnology, College of Life Science, Nanjing Normal University, Nanjing, People's Republic of China.
Xu
Haidong
H
Huang
Guodong
G
Chen
Ting
T
Liu
Guiyou
G
Chai
Nan
N
Ji
Yin
Y
Wang
Siyuan
S
Dai
Yijun
Y
Yuan
Sheng
S
D000192:Actinomycetales / Q000187:drug effects* / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects*; D000431:Ethanol / Q000633:toxicity; D005632:Fructose / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry; D005960:Glucosides / Q000378:metabolism; D007529:Isoflavones / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D005960:Glucosides; D007529:Isoflavones; D005632:Fructose; D000431:Ethanol; C033607:puerarin