TY - JOUR
T1 - Screening and characterization of an Agrobacterium tumefaciens mutant strain producing high level of coenzyme Q10
AU - Kim, Tae Su
AU - Yoo, Ji Hyun
AU - Kim, Sang Yong
AU - Pan, Cheol Ho
AU - Kalia, Vipin C.
AU - Kang, Yun Chan
AU - Lee, Jung Kul
N1 - Funding Information:
This work was supported by the Energy Efficiency & Resources Core Technology Program of the Korea Institute of Energy Technology Evaluation and Planning (KETEP), granted financial resource from the Ministry of Trade, Industry & Energy, Republic of Korea (201320200000420). This research was also supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT & Future Planning (2013R1A1A2012159).
Publisher Copyright:
© 2014 Elsevier Ltd. All rights reserved.
Copyright:
Copyright 2015 Elsevier B.V., All rights reserved.
PY - 2015/1
Y1 - 2015/1
N2 - A mutant of Agrobacterium tumefaciens (S02-13) producing high levels of coenzyme Q10 (CoQ10), was selected by high-throughput screening after repeated NTG mutagenesis. Mutant S02-13 was resistant to sodium azide and menadione and showed an eight-fold increase in CoQ10 production, as compared to that of the parent strain. The amount of CoQ10 produced by this mutant reached 350 mg l-1 in pH-stat fed-batch culture. Mutant S02-13 differed from wild-type in morphology, biochemical properties, and proteome profiles. Scanning electron microscopy results revealed a remarkable change in morphology: the wild-type strain had a rod shape, whereas the mutant S02-13 was coccoid. In contrast to wild-type, the mutant S02-13 strain showed a negative reaction to tryptophan deaminase and N2 reduction, and a positive reaction to nitrate production in API kit assays. The spots representing proteins that were increased in the proteome expression of the mutant S02-13 strain were identified as glyceraldehyde-3-phosphate dehydrogenase (GAPDH), glucosaminitol dehydrogenase (GlcNOH), and superoxide dismutase (SOD). In particular, the enzyme activities of GAPDH and GlcNOH increased eight-fold and three-fold, respectively, in the mutant, as compared to the wild-type.
AB - A mutant of Agrobacterium tumefaciens (S02-13) producing high levels of coenzyme Q10 (CoQ10), was selected by high-throughput screening after repeated NTG mutagenesis. Mutant S02-13 was resistant to sodium azide and menadione and showed an eight-fold increase in CoQ10 production, as compared to that of the parent strain. The amount of CoQ10 produced by this mutant reached 350 mg l-1 in pH-stat fed-batch culture. Mutant S02-13 differed from wild-type in morphology, biochemical properties, and proteome profiles. Scanning electron microscopy results revealed a remarkable change in morphology: the wild-type strain had a rod shape, whereas the mutant S02-13 was coccoid. In contrast to wild-type, the mutant S02-13 strain showed a negative reaction to tryptophan deaminase and N2 reduction, and a positive reaction to nitrate production in API kit assays. The spots representing proteins that were increased in the proteome expression of the mutant S02-13 strain were identified as glyceraldehyde-3-phosphate dehydrogenase (GAPDH), glucosaminitol dehydrogenase (GlcNOH), and superoxide dismutase (SOD). In particular, the enzyme activities of GAPDH and GlcNOH increased eight-fold and three-fold, respectively, in the mutant, as compared to the wild-type.
KW - 2-Dimensional electrophoresis
KW - Coenzyme Q
KW - Mutagenesis
KW - Production
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U2 - 10.1016/j.procbio.2014.10.024
DO - 10.1016/j.procbio.2014.10.024
M3 - Article
AN - SCOPUS:84920839551
VL - 50
SP - 33
EP - 39
JO - Process Biochemistry
JF - Process Biochemistry
SN - 1359-5113
IS - 1
ER -