Characterization of beta-carotene ketolases, CrtW, from marine bacteria by complementation analysis in Escherichia coli.
A complementation analysis was performed in Escherichia coli to evaluate the efficiency of beta-carotene ketolases (CrtW) from the marine bacteria Brevundimonas sp. SD212, Paracoccus sp. PC1 (Alcaligenes PC-1), and Paracoccus sp. N81106 (Agrobacterium aurantiacum), for astaxanthin production. Each crtW gene was expressed in Escherichia coli synthesizing zeaxanthin due to the presence of plasmid pACCAR25DeltacrtX. Carotenoids that accumulated in the resulting E. coli transformants were examined by chromatographic and spectroscopic analyses. The transformant carrying the Paracoccus sp. PC1 or N81106 crtW gene accumulated high levels of adonixanthin, which is the final astaxanthin precursor for CrtW, and astaxanthin, while the E. coli transformant with crtW from Brevundimonas sp. SD212 did not accumulate any adonixanthin and produced a high level of astaxanthin. These results show efficient conversion by CrtW of Brevundimonas sp. SD212 from adonixanthin to astaxanthin, which is a new-found characteristic of a bacterial CrtW enzyme. The phylogenetic positions between CrtW of the two genera, Brevundimonas and Paracoccus, are distant, although they fall into alpha-Proteobacteria.
Choi
Seon-kang
SK
Laboratory of Applied Molecular Design, Marine Biotechnology Institute, Heita, Kamaishi-shi, Iwate 026-0001, Japan.
Nishida
Yasuhiro
Y
Matsuda
Satoru
S
Adachi
Kyoko
K
Kasai
Hiroaki
H
Peng
Xue
X
Komemushi
Sadao
S
Miki
Wataru
W
Misawa
Norihiko
N
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000145:classification / Q000235:genetics*; D002338:Carotenoids / Q000032:analysis / Q000378:metabolism*; D042063:Caulobacteraceae / Q000145:classification / Q000201:enzymology / Q000235:genetics*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005816:Genetic Complementation Test; D005822:Genetic Vectors; D010105:Oxygenases / Q000096:biosynthesis* / Q000145:classification / Q000235:genetics*; D010230:Paracoccus / Q000145:classification / Q000201:enzymology / Q000235:genetics*; D010802:Phylogeny; D020539:Sequence Analysis, Protein; D024341:Xanthophylls / Q000032:analysis / Q000096:biosynthesis
D001426:Bacterial Proteins; D017931:DNA Primers; D024341:Xanthophylls; D002338:Carotenoids; C005948:astaxanthine; D010105:Oxygenases; C093259:beta-carotene ketolase