Proteome analyses of hydrogen-producing hyperthermophilic archaeon Thermococcus onnurineus NA1 in different one-carbon substrate culture conditions.
Thermococcus onnurineus NA1, a sulfur-reducing hyperthermophilic archaeon, is capable of H(2)-producing growth, considered to be hydrogenogenic carboxydotrophy. Utilization of formate as a sole energy source has been well studied in T. onnurineus NA1. However, whether formate can be used as its carbon source remains unknown. To obtain a global view of the metabolic characteristics of H(2)-producing growth, a quantitative proteome analysis of T. onnurineus NA1 grown on formate, CO, and starch was performed by combining one-dimensional SDS-PAGE with nano UPLC-MS(E). A total of 587 proteins corresponding to 29.7% of the encoding genes were identified, and the major metabolic pathways (especially energy metabolism) were characterized at the protein level. Expression of glycolytic enzymes was common but more highly induced in starch-grown cells. In contrast, enzymes involved in key steps of the gluconeogenesis and pentose phosphate pathways were strongly up-regulated in formate-grown cells, suggesting that formate could be utilized as a carbon source by T. onnurineus NA1. In accordance with the genomic analysis, comprehensive proteomic analysis also revealed a number of hydrogenase clusters apparently associated with formate metabolism. On the other hand, CODH and CO-induced hydrogenases belonging to the Hyg4-II cluster, as well as sulfhydrogenase-I and Mbx, were prominently expressed during CO culture. Our data suggest that CO can be utilized as a sole energy source for H(2) production via an electron transport mechanism and that CO(2) produced from catabolism or CO oxidation by CODH and CO-induced hydrogenases may subsequently be assimilated into the organic carbon. Overall, proteomic comparison of formate- and CO-grown cells with starch-grown cells revealed that a single carbon compound, such as formate and CO, can be utilized as an efficient substrate to provide cellular carbon and/or energy by T. onnurineus NA1.
Division of Life Science, Korea Basic Science Institute, Daejeon 305-806, Republic of Korea.
Yoon-Jung
YJ
Moon
Joseph
J
Kwon
Sung-Ho
SH
Yun
Hye Li
HL
Lim
Min-Sik
MS
Kim
Sung Gyun
SG
Kang
Jung-Hyun
JH
Lee
Jong-Soon
JS
Choi
Seung Il
SI
Kim
Young-Ho
YH
Chung
D000220:Adaptation, Biological; D000596:Amino Acids / Q000378:metabolism; D019843:Archaeal Proteins / Q000378:metabolism*; D050260:Carbohydrate Metabolism; D002248:Carbon Monoxide / Q000378:metabolism; D003470:Culture Media; D005561:Formates / Q000378:metabolism; D006859:Hydrogen / Q000378:metabolism; D006864:Hydrogenase / Q000235:genetics / Q000378:metabolism; D053858:Metabolic Networks and Pathways; D005810:Multigene Family; D020543:Proteome / Q000378:metabolism*; D013213:Starch / Q000378:metabolism; D013312:Stress, Physiological; D019712:Thermococcus / Q000254:growth & development / Q000378:metabolism* / Q000502:physiology
D000596:Amino Acids; D019843:Archaeal Proteins; D003470:Culture Media; D005561:Formates; D020543:Proteome; C030544:formic acid; D002248:Carbon Monoxide; D006859:Hydrogen; D013213:Starch; D006864:Hydrogenase