TY - JOUR
T1 - E. coli biosensor based on modular GFP and luxI/luxR cyclic amplification circuit for sensitive detection of lysine
AU - Wang, Wenjia
AU - Zhang, Jing
AU - Tao, Hui
AU - Lv, Xuefei
AU - Deng, Yulin
AU - Li, Xiaoqiong
N1 - Publisher Copyright:
© 2022, Springer-Verlag GmbH Germany, part of Springer Nature.
PY - 2022/12
Y1 - 2022/12
N2 - In this study, an E. coli biosensor based on modular green fluorescent protein and luxI/IuxR cycle amplification circuit was constructed for sensitive detection of bioavailable lysine. The results indicated that the luxI/IuxR positive feedback circuit based on quorum sensing can be used as a signal amplifier to improve the sensitivity to lysine detection with the detection limit of 256 nM. The presented method was more sensitive than the previously reported whole-cell fluorescent microbial biosensors. In addition, the developed E. coli biosensor was specific for lysine detection, and other amino acids and proteins did not cause any interference. The constructed genetic engineered biosensor was accurate for lysine detection, the lysine content of 6.87 ± 0.36% in tryptone was successfully measured, and after adding 10, 30, and 50 μM lysine in tryptone, the recoveries of 109.98 ± 10.44%, 103.88 ± 7.66%, and 105.89 ± 6.34% were obtained, respectively. Furthermore, as the design of the genetic engineered biosensor is modular, it can conceivably be utilized as a component in the design of more complex synthetic gene circuits without any changes to the amplifier and reporter system. Graphical abstract: [Figure not available: see fulltext.]
AB - In this study, an E. coli biosensor based on modular green fluorescent protein and luxI/IuxR cycle amplification circuit was constructed for sensitive detection of bioavailable lysine. The results indicated that the luxI/IuxR positive feedback circuit based on quorum sensing can be used as a signal amplifier to improve the sensitivity to lysine detection with the detection limit of 256 nM. The presented method was more sensitive than the previously reported whole-cell fluorescent microbial biosensors. In addition, the developed E. coli biosensor was specific for lysine detection, and other amino acids and proteins did not cause any interference. The constructed genetic engineered biosensor was accurate for lysine detection, the lysine content of 6.87 ± 0.36% in tryptone was successfully measured, and after adding 10, 30, and 50 μM lysine in tryptone, the recoveries of 109.98 ± 10.44%, 103.88 ± 7.66%, and 105.89 ± 6.34% were obtained, respectively. Furthermore, as the design of the genetic engineered biosensor is modular, it can conceivably be utilized as a component in the design of more complex synthetic gene circuits without any changes to the amplifier and reporter system. Graphical abstract: [Figure not available: see fulltext.]
KW - Amplification
KW - Biosensor
KW - LuxI
KW - LuxR
KW - Lysine
UR - http://www.scopus.com/inward/record.url?scp=85139957968&partnerID=8YFLogxK
U2 - 10.1007/s00216-022-04364-1
DO - 10.1007/s00216-022-04364-1
M3 - Article
C2 - 36253476
AN - SCOPUS:85139957968
SN - 1618-2642
VL - 414
SP - 8299
EP - 8307
JO - Analytical and Bioanalytical Chemistry
JF - Analytical and Bioanalytical Chemistry
IS - 29-30
ER -