Proteomic analysis on acetate metabolism in Citrobacter sp. BL-4.
Mass production of glucosamine (GlcN) using microbial cells is a worthy approach to increase added values and keep safety problems in GlcN production process. Prior to set up a microbial cellular platform, this study was to assess acetate metabolism in Citrobacter sp. BL-4 (BL-4) which has produced a polyglucosamine PGB-2. The LC-MS analysis was conducted after protein separation on the 1D-PAGE to accomplish the purpose of this study. 280 proteins were totally identified and 188 proteins were separated as acetate-related proteins in BL-4. Acetate was converted to acetyl-CoA by acetyl-CoA synthetase up-regulated in the acetate medium. The glyoxylate bypass in the acetate medium was up-regulated with over-expression of isocitrate lyases and 2D-PAGE confirmed this differential expression. Using (1)H-NMR analysis, the product of isocitrate lyases, succinate, increased about 15 times in the acetate medium. During acetate metabolism proteins involved in the lipid metabolism and hexosamine biosynthesis were over-expressed in the acetate medium, while proteins involved in TCA cycle, pentose phosphate cycle and purine metabolism were down-regulated. Taken together, the results from the proteomic analysis can be applied to improve GlcN production and to develop metabolic engineering in BL-4.
Department of Food Science and Nutrition, Dong-eui University, Busan 614-714, Republic of Korea.
Young-Man
YM
Kim
Sung-Eun
SE
Lee
Byeoung-Soo
BS
Park
Mi-Kyung
MK
Son
Young-Mi
YM
Jung
Seung-Ok
SO
Yang
Hyung-Kyoon
HK
Choi
Sung-Ho
SH
Hur
Jong Hwa
JH
Yum
D000085:Acetates / Q000378:metabolism*; D000105:Acetyl Coenzyme A / Q000378:metabolism; D057005:Bioengineering; D002853:Chromatography, Liquid; D002952:Citric Acid Cycle; D002954:Citrobacter / Q000254:growth & development / Q000378:metabolism*; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D015964:Gene Expression Regulation, Bacterial; D005944:Glucosamine / Q000378:metabolism; D007522:Isocitrate Lyase / Q000378:metabolism; D050356:Lipid Metabolism; D013058:Mass Spectrometry; D019906:Nuclear Magnetic Resonance, Biomolecular; D010427:Pentose Phosphate Pathway; D040901:Proteomics; D011687:Purines / Q000378:metabolism; D019802:Succinic Acid / Q000378:metabolism
D000085:Acetates; D003470:Culture Media; D011687:Purines; D000105:Acetyl Coenzyme A; D019802:Succinic Acid; D007522:Isocitrate Lyase; D005944:Glucosamine; C030985:purine