Reconstruction of the carotenoid biosynthetic pathway of Cronobacter sakazakii BAA894 in Escherichia coli.
Cronobacter sakazakii could form yellow-pigmented colonies. However, the chemical structure and the biosynthetic pathway of the yellow pigments have not been identified. In this study, the yellow pigments of C. sakazakii BAA894 were purified and analyzed. The major components of the yellow pigments were confirmed as zeaxanthin-monoglycoside and zeaxanthin-diglycoside. A gene cluster containing seven genes responsible for the yellow pigmentation in C. sakazakii BAA894 was identified. The seven genes of C. sakazakii BAA894 or parts of them were reconstructed in a heterologous host Escherichia coli DH5α. The pigments formed in these E. coli strains were isolated and analyzed by thin layer chromatography, UV-visible spectroscopy, high performance liquid chromatography, and electron spray ionization-mass spectrometry. These redesigned E. coli strains could produce different carotenoids. E. coli strain expressing all the seven genes could produce zeaxanthin-monoglycoside and zeaxanthin-diglycoside; E. coli strains expressing parts of the seven genes could produce lycopene, β-carotene, cryptoxanthin or zeaxanthin. This study identified the gene cluster responsible for the yellow pigmentation in C. sakazakii BAA894.
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, China ; Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China.
Wei
W
Zhang
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, China ; Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, Wuxi, China.
Xiaoqing
X
Hu
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, China.
Liqin
L
Wang
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, China ; Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, Wuxi, China.
Xiaoyuan
X
Wang
D053898:Biosynthetic Pathways / Q000235:genetics*; D002338:Carotenoids / Q000032:analysis / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D002855:Chromatography, Thin Layer; D003001:Cloning, Molecular; D044083:Cronobacter sakazakii / Q000235:genetics / Q000378:metabolism*; D065366:Cryptoxanthins; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D005798:Genes, Bacterial; D000077276:Lycopene; D005810:Multigene Family; D010860:Pigments, Biological / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D021241:Spectrometry, Mass, Electrospray Ionization; D013056:Spectrophotometry, Ultraviolet; D024341:Xanthophylls / Q000378:metabolism; D065146:Zeaxanthins; D019207:beta Carotene / Q000378:metabolism
D065366:Cryptoxanthins; D004269:DNA, Bacterial; D010860:Pigments, Biological; D024341:Xanthophylls; D065146:Zeaxanthins; D019207:beta Carotene; D002338:Carotenoids; D000077276:Lycopene