The glyoxylate shunt is essential for CO2-requiring oligotrophic growth of Rhodococcus erythropolis N9T-4

Verfasser / Beitragende:
[Takanori Yano, Nobuyuki Yoshida, Fujio Yu, Miki Wakamatsu, Hiroshi Takagi]
Ort, Verlag, Jahr:
2015
Enthalten in:
Applied Microbiology and Biotechnology, 99/13(2015-07-01), 5627-5637
Format:
Artikel (online)
ID: 605505861
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024 7 0 |a 10.1007/s00253-015-6500-x  |2 doi 
035 |a (NATIONALLICENCE)springer-10.1007/s00253-015-6500-x 
245 0 4 |a The glyoxylate shunt is essential for CO2-requiring oligotrophic growth of Rhodococcus erythropolis N9T-4  |h [Elektronische Daten]  |c [Takanori Yano, Nobuyuki Yoshida, Fujio Yu, Miki Wakamatsu, Hiroshi Takagi] 
520 3 |a Rhodococcus erythropolis N9T-4 shows extremely oligotrophic growth requiring atmospheric CO2 and forms its colonies on an inorganic basal medium (BM) without any additional carbon source. Screening of a random mutation library constructed by a unique genome deletion method that we established indicated that the aceA, aceB, and pckG genes encoding isocitrate lyase, malate synthase, and phosphoenolpyruvate carboxykinase, respectively, were requisite for survival on BM plates. The aceA- and aceB deletion mutants and the pckG deletion mutant grew well on BM plates containing L-malate and D-glucose, respectively, suggesting that the glyoxylate (GO) shunt and gluconeogenesis are essential for the oligotrophic growth of N9T-4. Interestingly, most of the enzyme activities in the TCA cycle were observed in the cell-free extract of N9T-4, with perhaps the most important exception being α-ketoglutarate dehydrogenase (KGDH) activity. Instead of the KGDH activity, we detected a remarkable level of α-ketoglutarate decarboxylase (KGD) activity, which is the activity exhibited by the E1 component of the KGDH complex in Mycobacterium tuberculosis. The recombinant KGD of N9T-4 catalyzed the decarboxylation of α-ketoglutarate to form succinic semialdehyde (SSA) in a time-dependent manner. Since N9T-4 also showed a detectable SSA dehydrogenase activity, we concluded that N9T-4 possesses a variant TCA cycle, which uses SSA rather than succinyl-CoA. These results suggest that oligotrophic N9T-4 cells utilize the GO shunt to avoid the loss of carbons as CO2 and to conserve CoA units in the TCA cycle. 
540 |a Springer-Verlag Berlin Heidelberg, 2015 
690 7 |a Random mutagenesis  |2 nationallicence 
690 7 |a Carbon dioxide  |2 nationallicence 
690 7 |a Glyoxylate shunt  |2 nationallicence 
690 7 |a TCA cycle  |2 nationallicence 
690 7 |a Oligotroph  |2 nationallicence 
690 7 |a Rhodococcus erythropolis N9T-4  |2 nationallicence 
700 1 |a Yano  |D Takanori  |u Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, 630-0192, Ikoma, Nara, Japan  |4 aut 
700 1 |a Yoshida  |D Nobuyuki  |u Department of Applied Chemistry and Biochemical Engineering, Graduate school of Engineering, Shizuoka University, 3-5-1 Johoku, Naka-ku, 432-8561, Hamamatsu, Japan  |4 aut 
700 1 |a Yu  |D Fujio  |u Yokohama Research Labolatories, Mitsubishi Rayon Co., Ltd., 10-1, Daikoku-cho, Tsurumi-ku, 230-0053, Yokohama, Japan  |4 aut 
700 1 |a Wakamatsu  |D Miki  |u Yokohama Research Labolatories, Mitsubishi Rayon Co., Ltd., 10-1, Daikoku-cho, Tsurumi-ku, 230-0053, Yokohama, Japan  |4 aut 
700 1 |a Takagi  |D Hiroshi  |u Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, 630-0192, Ikoma, Nara, Japan  |4 aut 
773 0 |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/13(2015-07-01), 5627-5637  |x 0175-7598  |q 99:13<5627  |1 2015  |2 99  |o 253 
856 4 0 |u https://doi.org/10.1007/s00253-015-6500-x  |q text/html  |z Onlinezugriff via DOI 
898 |a BK010053  |b XK010053  |c XK010000 
900 7 |a Metadata rights reserved  |b Springer special CC-BY-NC licence  |2 nationallicence 
908 |D 1  |a research-article  |2 jats 
949 |B NATIONALLICENCE  |F NATIONALLICENCE  |b NL-springer 
950 |B NATIONALLICENCE  |P 856  |E 40  |u https://doi.org/10.1007/s00253-015-6500-x  |q text/html  |z Onlinezugriff via DOI 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Yano  |D Takanori  |u Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, 630-0192, Ikoma, Nara, Japan  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Yoshida  |D Nobuyuki  |u Department of Applied Chemistry and Biochemical Engineering, Graduate school of Engineering, Shizuoka University, 3-5-1 Johoku, Naka-ku, 432-8561, Hamamatsu, Japan  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Yu  |D Fujio  |u Yokohama Research Labolatories, Mitsubishi Rayon Co., Ltd., 10-1, Daikoku-cho, Tsurumi-ku, 230-0053, Yokohama, Japan  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Wakamatsu  |D Miki  |u Yokohama Research Labolatories, Mitsubishi Rayon Co., Ltd., 10-1, Daikoku-cho, Tsurumi-ku, 230-0053, Yokohama, Japan  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Takagi  |D Hiroshi  |u Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, 630-0192, Ikoma, Nara, Japan  |4 aut 
950 |B NATIONALLICENCE  |P 773  |E 0-  |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/13(2015-07-01), 5627-5637  |x 0175-7598  |q 99:13<5627  |1 2015  |2 99  |o 253