Engineering a vitamin B12 high-throughput screening system by riboswitch sensor in Sinorhizobium meliloti

BMC Biotechnology
Yingying CaiDawei Zhang

Abstract

As a very important coenzyme in the cell metabolism, Vitamin B12 (cobalamin, VB12) has been widely used in food and medicine fields. The complete biosynthesis of VB12 requires approximately 30 genes, but overexpression of these genes did not result in expected increase of VB12 production. High-yield VB12-producing strains are usually obtained by mutagenesis treatments, thus developing an efficient screening approach is urgently needed. By the help of engineered strains with varied capacities of VB12 production, a riboswitch library was constructed and screened, and the btuB element from Salmonella typhimurium was identified as the best regulatory device. A flow cytometry high-throughput screening system was developed based on the btuB riboswitch with high efficiency to identify positive mutants. Mutation of Sinorhizobium meliloti (S. meliloti) was optimized using the novel mutation technique of atmospheric and room temperature plasma (ARTP). Finally, the mutant S. meliloti MC5-2 was obtained and considered as a candidate for industrial applications. After 7 d's cultivation on a rotary shaker at 30 °C, the VB12 titer of S. meliloti MC5-2 reached 156 ± 4.2 mg/L, which was 21.9% higher than that of the wild type strain S. meliloti...Continue Reading

References

Jul 24, 2001·Briefings in Bioinformatics·M S GelfandA A Mironov
Oct 7, 2003·Annual Review of Biochemistry·Ruma Banerjee, Stephen W Ragsdale
Dec 31, 2003·Trends in Genetics : TIG·Alexey G VitreschakMikhail S Gelfand
Jan 6, 2004·Nucleic Acids Research·Ali NahviRonald R Breaker
Feb 22, 2005·Nature Biotechnology·Travis S Bayer, Christina D Smolke
Mar 10, 2006·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Kenneth H Wolfe
Aug 4, 2006·Applied and Environmental Microbiology·Allyson M MacLeanTurlough M Finan
May 8, 2007·Journal of Bacteriology·Jorge C Escalante-Semerena
Dec 18, 2008·Current Opinion in Microbiology·Hervé TettelinDuccio Medini
Sep 26, 2009·Chembiochem : a European Journal of Chemical Biology·Guangyu Yang, Stephen G Withers
May 11, 2010·Nature Chemical Biology·Joy SinhaJustin P Gallivan
Jul 28, 2010·Chemistry & Biology·Casey C FowlerYingfu Li
Aug 12, 2010·Molecular Plant-microbe Interactions : MPMI·Michiko E Taga, Graham C Walker
Nov 26, 2010·Cold Spring Harbor Perspectives in Biology·Ronald R Breaker
Feb 15, 2011·Nature Structural & Molecular Biology·Peter Y Watson, Martha J Fedor
Sep 20, 2011·Molecular Cell·Ronald R Breaker
Jan 22, 2013·Cell·Alexander Serganov, Evgeny Nudler
Jun 11, 2016·Bioprocess and Biosystems Engineering·Huina DongJibin Sun

❮ Previous
Next ❯

Citations

Jan 11, 2020·Applied Microbiology and Biotechnology·Qinghui YuGuoquan Hu
Dec 15, 2019·Trends in Biotechnology·Emily K Bowman, Hal S Alper
May 1, 2021·International Journal of Molecular Sciences·Larissa BalabanovaLiudmila Tekutyeva
May 1, 2021·International Journal of Molecular Sciences·Marta IrlaTrygve Brautaset
Jul 6, 2021·Frontiers in Bioengineering and Biotechnology·Yanyan WangDawei Zhang

❮ Previous
Next ❯

Methods Mentioned

BETA
biosensors
biosensor
biosens
FACS
PCR
flow cytometry
electrophoresis
Fluorescence Activated Cell Sorting

Software Mentioned

btuB
SY

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.