title: Constitutive expression of Campylobacter jejuni truncated hemoglobin CtrHb improves the growth of Escherichia coli cell under aerobic and anaerobic conditions.
abstract: Bacteria hemoglobin could bind to the oxygen, transfer it from the intracellular microenvironment to the respiration process and sustain the energy for the metabolism and reproduction of cells. Heterologous expression of bacteria hemoglobin gene could improve the capacity of the host on oxygen-capturing and allow it to grow even under microaerophilic condition. To develop a system based on hemoglobin to help bacteria cells overcome the oxygen shortage in fermentation, in this study, Campylobacter jejuni truncated hemoglobin (CtrHb) gene was synthesized and expressed under the control of constitutive expression promoters P2 and P(SPO1-II) in Escherichia coli. As showed by the growth curves of the two recombinants P2-CtrHb and P(SPO1-II)-CtrHb, constitutive expression of CtrHb improved cell growth under aerobic shaking-flasks, anaerobic capped-bottles and bioreactor conditions. According to the NMR analysis, this improvement might come from the expression of hemoglobin which could boost the metabolism of cells by supplying more oxygen to the respiratory chain processes. Through semi-quantitative RT-PCR and CO differential spectrum assays, we further discussed the connection between the growth patterns of the recombinants, the expression level of CtrHb and oxygen binding capacity of CtrHb in cells. Based on the growth patterns of these recombinants in bioreactor, a possible choice on different type of recombinants under specific fermentation conditions was also suggested in this study.
authors.affiliation: College of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan 430023, China. Electronic address: jiangke.yang@gmail.com.
authors.forename: Jiang-Ke
authors.initials: JK
authors.lastname: Yang
authors.affiliation: College of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan 430023, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Xiong
authors.affiliation: College of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan 430023, China.
authors.forename: Li
authors.initials: L
authors.lastname: Xu
authors.affiliation: College of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan 430023, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Li
authors.affiliation: College of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan 430023, China.
authors.forename: Xiu-Ju
authors.initials: XJ
authors.lastname: Zhao
mesh_terms: D000332:Aerobiosis; D000595:Amino Acid Sequence; D000693:Anaerobiosis; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D019149:Bioreactors / Q000382:microbiology; D016123:Campylobacter jejuni / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D015870:Gene Expression; D005798:Genes, Bacterial; D007218:Industrial Microbiology; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D010100:Oxygen / Q000378:metabolism; D010802:Phylogeny; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D054793:Truncated Hemoglobins / Q000235:genetics* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D011994:Recombinant Proteins; D054793:Truncated Hemoglobins; D010100:Oxygen
keywords: CO differential display; Constitutive promoter; CtrHb; Growth curve; Semi-quantitative PCR