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A Novel Biosensor to Advance Diverse High-Level Production of Microbial Cell Factories
A research group at KAIST presented a novel biosensor which can produce diverse, high-level microbial cell factories. The biosensor monitors the concentration of products and even intermediates when new strains are being developed. This strategy provides a new platform for manufacturing diverse natural products from renewable resources. The team succeeded in creating four natural products of high-level pharmaceutical importance with this strategy.
Malonyl-CoA is a major building block for many value-added chemicals including diverse natural products with pharmaceutical importance. However, due to the low availability of malonyl-CoA in bacteria, many malonyl-CoA-derived natural products have been produced by chemical synthesis or extraction from natural resources that are harmful to the environment and are unsustainable. For the sustainable biological production of malonyl-CoA-derived natural products, increasing the intracellular malonyl-CoA pool is necessary. To this end, the development of a robust and efficient malonyl-CoA biosensor was required to monitor the concentration of intracellular malonyl-CoA abundance as new strains are developed.
Metabolic engineering researchers at KAIST addressed this issue. This research reports the development of a simple and robust malonyl-CoA biosensor by repurposing a type III polyketide synthase (also known as RppA), which produces flaviolin, a colorimetric indicator of malonyl-CoA. Subsequently, the RppA biosensor was used for the rapid and efficient colorimetric screening of gene manipulation targets enabling enhanced malonyl-CoA abundance. The screened beneficial gene targets were employed for the high-level production of four representative natural products derived from malonyl-CoA. Compared with the previous strategies, which were expensive and time-consuming, the new biosensor could be easily applied to industrially relevant bacteria including Escherichia coli, Pseudomonas putida, and Corynebacterium glutamicum to enable a one-step process.
The study employs synthetic small regulatory RNA (sRNA) technology to rapidly and efficiently reduce endogenous target gene expression for improved malonyl-CoA production. The researchers constructed an E. coli genome-scale synthetic sRNA library targeting 1,858 genes covering all major metabolic genes in E. coli. This library was employed with the RppA biosensor to screen for gene targets which are believed to be beneficial for enhancing malonyl-CoA accumulation upon their expression knockdown.
From this colorimetric screening, 14 gene targets were selected, all of which were successful at significantly increasing the production of four natural products (6-methylsalicylic acid, aloesone, resveratrol, and naringenin). Although specific examples are demonstrated in E. coli as a host, the researchers showed that the biosensor is also functional in P. putida and C. glutamicum, industrially important representative gram-negative and gram-positive bacteria, respectively. The malonyl-CoA biosensor developed in this research will serve as an efficient platform for the rapid development of strains capable of producing natural products crucial for the pharmaceutical, chemical, cosmetics, and food industries.
An important aspect of this work is that the high-performance strains constructed in this research were developed rapidly and easily by utilizing the simple approach of colorimetric screening, without involving extensive metabolic engineering approaches. 6-Methylsalicylic acid (an antibiotic) could be produced to the highest titer reported for E. coli, and the microbial production of aloesone (a precursor of aloesin, an anti-inflammatory agent/whitening agent) was achieved for the first time.
“A sustainable process for producing diverse natural products using renewable resources is of great interest. This study represents the development of a robust and efficient malonyl-CoA biosensor generally applicable to a wide range of industrially important bacteria. The capability of this biosensor for screening a large library was demonstrated to show that the rapid and efficient construction of high-performance strains is feasible. This research will be useful for further accelerating the development process of strains capable of producing valuable chemicals to industrially relevant levels,” said Distinguished Professor Sang Yup Lee of the Department of Chemical and Biomolecular Engineering, who led the research.
This study entitled “Repurposing type III polyketide synthase as a malonyl-CoA biosensor for metabolic engineering in bacteria,” was published in the Proceedings of the National Academy of Sciences of the United States of America (PNAS) on October 02.
PhD students Dongsoo Yang and Won Jun Kim, MS student Shin Hee Ha, research staff Mun Hee Lee, Research Professor Seung Min Yoo, and Distinguished Professor Sang Yup Lee of the Department of Chemical and Biomolecular Engineering and Dr. Jong Hyun Choi of the Applied Microbiology Research Center at the Korea Research Institute of Bioscience and Biotechnology (KRIBB) participated in this research.
Figure: Type III polyketide synthase (RppA) as a malonyl-CoA biosensor. RppA converts five molecules of malonyl-CoA into one molecule of red-colored flaviolin. This schematic diagram shows the overall conceptualization of the malonyl-CoA biosensor by indicating that higher malonyl-CoA abundance leads to higher production and secretion of flaviolin, resulting in a deeper red color of the culture. This system was employed for the enhanced production of four representative natural products (6-methylsalicylic acid, aloesone, resveratrol, and naringenin) from engineered E. coli strains.
2018.10.11 View 10011 -
Permanent, Wireless Self-charging System Using NIR Band
(Professor Jung-Yong Lee from the Graduate School of Energy, Environment, Water and Sustainability)
As wearable devices are emerging, there are numerous studies on wireless charging systems. Here, a KAIST research team has developed a permanent, wireless self-charging platform for low-power wearable electronics by converting near-infrared (NIR) band irradiation to electrical energy. This novel technology can be applied to flexible, wearable charging systems without needing any attachments.
Colloidal-quantum-dots (CQDs) are promising materials for manufacturing semiconductors; in particular, PbS-based CQDs have facile optical tunability from the visible to infrared wavelength region. Hence, they can be applied to various devices, such as lighting, photovoltaics (PVs), and photodetectors.
Continuous research on CQD-based optoelectronic devices has increased their power conversion efficiency (PCE) to 12%; however, applicable fields have not yet been found for them. Meanwhile, wearable electronic devices commonly face the problem of inconvenient charging systems because users have to constantly charge batteries attached to an energy source.
A joint team led by Professor Jung-Yong Lee from the Graduate School of Energy, Environment, Water and Sustainability and Jang Wok Choi from Seoul National University decided to apply CQD PVs, which have high quantum efficiency in NIR band to self-charging systems on wearable devices.
They employed a stable and efficient NIR energy conversion strategy. The system was comprised of a PbS CQD-based PV module, a flexible interdigitated lithium-ion battery, and various types of NIR-transparent films.
The team removed the existing battery from the already commercialized wearable healthcare bracelet and replaced it with the proposed self-charging system. They confirmed that the system can be applied to a low power wearable device via the NIR band.
There have been numerous platforms using solar irradiation, but the newly developed platform has more advantages because it allows conventional devices to be much more comfortable to wear and charged easily in everyday life using various irradiation sources for constant charging.
With this aspect, the proposed platform facilitates more flexible designs, which are the important component for actual commercialization. It also secures higher photostability and efficient than existing structures.
Professor Lee said, “By using the NIR band, we proposed a new approach to solve charging system issues of wearable devices. I believe that this platform will be a novel platform for energy conversion and that its application can be further extended to various fields, including mobiles, IoTs, and drones.”
This research, led by PhD Se-Woong Baek and M.S. candidate Jungmin Cho, was published in Advanced Materials on May 11.
Figure 1. a) Conceptual NIR-driven self-charging system including a flexible CQD PVs module and an interdigitatedly structured LIB. b) Photographic images of a conventional wearable healthcare bracelet and a self-charging system-integrated wearable device.
Figure 2. Illustration of the CQD PVs structure and performance of the wireless self-charging platform.
2018.10.08 View 8560 -
AI ｜QC ITRC Opens at KAIST
(from left: Dean of College of Engineering Jong-Hwan Kim, Director of AI│QC ITRC June-Koo Rhee, Vice President for R&DB Heekyung Park and Director General for Industrial Policy Hong Taek Yong)
Artificial Intelligence｜The Quantum Computing Information Technology Research Center (AI｜QC ITRC) opened at KAIST on October 2.
AI｜QC ITRC, established with government funding, is the first institute specializing in quantum computing. Three universities (Seoul National University, Korea University, and Kyung Hee University), and four corporations, KT, Homomicus, Actusnetworks, and Mirae Tech are jointly participating in the center. Over four years, the institute will receive 3.2 billion KRW of research funds.
Last April, KAIST selected quantum technology as one of its flagship research areas. AI｜QC ITRC will dedicate itself to developing quantum computing technology that provides the computability required for human-level artificial intelligence. It will also foster leaders in related industries by introducing industry-academic educational programs in graduate schools.
QC is receiving a great deal of attention for transcending current digital computers in terms of computability. World-class IT companies like IBM, Google, and Intel and ventures including D-Wave, Rigetti, and IonQ are currently leading the industry and investing heavily in securing source technologies.
Starting from the establishment of the ITRC, KAIST will continue to plan strategies to foster the field of QC. KAIST will carry out two-track strategies; one is to secure source technology of first-generation QC technology, and the other is to focus on basic research that can preoccupy next-generation QC technology.
Professor June-Koo Rhee, the director of AI│QC ITRC said, “I believe that QC will be the imperative technology that enables the realization of the Fourth Industrial Revolution. AIQC ITRC will foster experts required for domestic academia and industries and build a foundation to disseminate the technology to industries.”
Vice President for R&DB Heekyung Park, Director General for Industrial Policy Hong Taek Yong from the Ministry of Science and ICT, Seung Pyo Hong from the Institute for Information & communications Technology Promotion, Head of Technology Strategy Jinhyon Youn from KT, and participating companies attended and celebrated the opening of the AI│QC ITRC.
2018.10.05 View 7055 -
Scientist of October, Professor Haeshin Lee
(Professor Haeshin Lee from the Department of Chemistry)
Professor Haeshin Lee from the Department of Chemistry received the ‘Science and Technology Award of October’ from the Ministry of Science and ICT and the National Research Foundation of Korea for his contribution to developing an antibleeding injection needle. This novel outcome will fundamentally prevent the problem of secondary infections of AIDS, Ebola and Hepatitis viruses transmitting from patients to medical teams.
This needle’s surface is coated with hemostatic materials. Its concept is simple and the key to this technology is to make materials that are firmly coated on the needle so that they can endure frictional force when being injected into skin and blood vessels. Moreover, the materials should be adhesive to skin and the interior of blood vessels, but harmless to humans.
Professor Lee found a solution from natural polymer ingredients. Catecholamine can be found in mussels. Professor Lee conjugated catechol groups on the chitosan backbone. He applied this mussel-inspired adhesive polymer Chitosan-catechol, which immediately forms an adhesive layer with blood, as a bioadhesion for the antibleeding injection needle.
Professor Lee said, “Chitosan-catechol, which copies the adhesive mechanism of mussels, shows high solubility in physiological saline as well as great mucoadhesion. Hence, it is perfectly suitable for coating the injection needle. Combining it with proteins allows for efficient drug delivery to the heart, which is a challenging injection location, so it will be also useful for treating incurable heart disease.”
2018.10.05 View 10615 -
Flexible Piezoelectric Acoustic Sensors for Speaker Recognition
A KAIST research team led by Professor Keon Jae Lee from the Department of Material Science and Engineering has developed a machine learning-based acoustic sensor for speaker recognition.
Acoustic sensors were spotlighted as one of the most intuitive bilateral communication devices between humans and machines. However, conventional acoustic sensors use a condenser-type device for measuring capacitance between two conducting layers, resulting in low sensitivity, short recognition distance, and low speaker recognition rates.
The team fabricated a flexible piezoelectric membrane by mimicking the basilar membrane in the human cochlear. Resonant frequencies vibrate corresponding regions of the trapezoidal piezoelectric membrane, which converts voice to electrical signal with a highly sensitive self-powered acoustic sensor.
This multi-channel piezoelectric acoustic sensor exhibits sensitivity more than two times higher and allows for more abundant voice information compared to conventional acoustic sensors, which can detect minute sounds from farther distances. In addition, the acoustic sensor can achieve a 97.5% speaker recognition rate using a machine learning algorithm, reducing by 75% error rate than the reference microphone.
AI speaker recognition is the next big thing for future individual customized services. However, conventional technology attempts to improve recognition rates by using software upgrades, resulting in limited speaker recognition rates. The team enhanced the speaker recognition system by replacing the existing hardware with an innovative flexible piezoelectric acoustic sensor. Further software improvement of the piezoelectric acoustic sensor will significantly increase the speaker and voice recognition rate in diverse environments.
Professor Lee said, “Highly sensitive self-powered acoustic sensors for speaker recognition can be used for personalized voice services such as smart home appliances, AI secretaries, always-on IoT, biometric authentication, and FinTech.”
These research “Basilar Membrane-Inspired Self-Powered Acoustic Sensor” and “Machine Learning-based Acoustic Sensor for Speaker Recognition” were published in the September 2018 issue of Nano Energy.
Firgure 1: A flexible piezoelectric acoustic sensor mimicking the human cochlear.
Figure 2: Speaker recognition with a machine learning algorithm.
2018.10.04 View 7665 -
The 1st Korea Toray Science and Technology Awardee, Prof. Sukbok Chang
(Distinguished Professor Sukbok Chang from the Department of Chemistry)
The Korea Toray Science Foundation (KTSF) awarded the first Korea Toray Science Technology Award in basic science to Distinguished Professor Sukbok Chang from the Department of Chemistry on September 19.
KTSF was established in January 2018, and its award goes to researchers who have significantly contributed to the development of chemistry and materials research with funds to support research projects.
Distinguished Professor Chang has devoted himself in organocatalysis research; in particular, his work on catalysts for effective lactam formation, which was an intricate problem, received great attention.
The award ceremony will take place in The Federation of Korean Industries Hall on October 31. KTFS board members, judges, and the CEO of Toray Industries Akihiro Nikkaku will attend the ceremony. Also, Dr. Ryoji Noyori, the Nobel Laureate in Chemistry, will give a talk on the role of chemistry and creative challenges as a researcher.
2018.10.04 View 8944 -
Spray Coated Tactile Sensor on a 3-D Surface for Robotic Skin
Robots will be able to conduct a wide variety of tasks as well as humans if they can be given tactile sensing capabilities.
A KAIST research team has reported a stretchable pressure insensitive strain sensor by using an all solution-based process. The solution-based process is easily scalable to accommodate for large areas and can be coated as a thin-film on 3-dimensional irregularly shaped objects via spray coating. These conditions make their processing technique unique and highly suitable for robotic electronic skin or wearable electronic applications.
The making of electronic skin to mimic the tactile sensing properties of human skin is an active area of research for various applications such as wearable electronics, robotics, and prosthetics. One of the major challenges in electronic skin research is differentiating various external stimuli, particularly between strain and pressure. Another issue is uniformly depositing electrical skin on 3-dimensional irregularly shaped objects.
To overcome these issues, the research team led by Professor Steve Park from the Department of Materials Science and Engineering and Professor Jung Kim from the Department of Mechanical Engineering developed electronic skin that can be uniformly coated on 3-dimensional surfaces and distinguish mechanical stimuli. The new electronic skin can also distinguish mechanical stimuli analogous to human skin. The structure of the electronic skin was designed to respond differently under applied pressure and strain. Under applied strain, conducting pathways undergo significant conformational changes, considerably changing the resistance. On the other hand, under applied pressure, negligible conformational change in the conducting pathway occurs; e-skin is therefore non-responsive to pressure. The research team is currently working on strain insensitive pressure sensors to use with the developed strain sensors.
The research team also spatially mapped the local strain without the use of patterned electrode arrays utilizing electrical impedance tomography (EIT). By using EIT, it is possible to minimize the number of electrodes, increase durability, and enable facile fabrication onto 3-dimensional surfaces.
Professor Park said, “Our electronic skin can be mass produced at a low cost and can easily be coated onto complex 3-dimensional surfaces. It is a key technology that can bring us closer to the commercialization of electronic skin for various applications in the near future.”
The result of this work entitled “Pressure Insensitive Strain Sensor with Facile Solution-based Process for Tactile Sensing Applications” was published in the August issue of ACS Nano as a cover article.
(Figure: Detecting mechanical stimuli using electrical impedance tomography.)
2018.09.21 View 8656 -
President Shin Presents Opportunities & Challenges of the 4IR at the Summer Davos Forum
(President Shin makes a keynote speech at the 2018 Summer Davos Forum in China on Sept.20.)
KAIST co-hosted the Asia Session with the World Economic Forum during the 2018 Summer Davos Forum in Tianjin, China from September 18 through 20. The session highlighted regional collaboration in Asia to promote inclusive growth in the Fourth Industrial Revolution.
KAIST is working closely with the WEF to take the lead in the Fourth Industrial Revolution. Last July, KAIST established the Fourth Industrial Revolution Information Center (FIRIC) at the KAIST Institute and signed an MOU with the Center for the Fourth Industrial Revolution (C4IR) at the WEF in October. The session is a follow-up event KAIST and the C4IR agreed to last year during the Roundtable Session held in Seoul.
Many experts in new emerging industries as well as many project directors, including Director Murat Sonmez of the C4IR, attended the session KAIST hosted. Director Chizuru Suga at the C4IR in Japan, Director Danil Kerimi in China, and Director Shailesh Sharda in India also attended the session and discussed ways to expand collaboration and networks among the countries.
In his keynote speech at the session on September 20, President Sung-Chul Shin presented how the Korean government is trying to drive the economy by strategically investing in focused industries in the new global industrial environment. President Shin introduced the government’s strategic roadmap to build the competitiveness of emerging technologies such as AI, blockchain, and precision medicine.
He also stressed that the three components of innovation, collaboration, and speed should be prioritized in all sectors for the successful realization of the Fourth Industrial Revolution. For instance, innovation in education, research, and technology commercialization, expansive domestic and international collaboration beyond the private and public sectors, speedy deregulation, and efficient governance will all be critical.
He also said that KAIST will launch new pilot collaboration projects along with the WEF soon. “We paved the way for leading the network with major countries including Japan and India for advancing the Fourth Industrial Revolution through this session,” President Shin said.
2018.09.21 View 8536 -
KAIST Develops VRFB with Longer Durability
(from left: PhD candidate Soohyun Kim, Professor Hee-Tak Kim and PhD candidate Junghoon Choi)
There has been growing demand for large-scale storage for energy produced from renewable energy sources in an efficient and stable way. To meet this demand, a KAIST research team developed a new vanadium redox-flow battery (VRFB) with 15 times greater capacity retention and five times longer durability. This VRFB battery can be an excellent candidate for a large-scale rechargeable battery with no risk of explosion.
The VRFB has received much attention for its high efficiency and reliability with the absence of cross-contamination. However, it has the limitation of having insufficient charge and discharge efficiency and a low capacity retention rate because its perfluorinated membrane is very permeable to any active materials. To minimize energy loss, it needs a membrane that has low vanadium ion permeability and high ion conductivity. Hence, there was an attempt to incorporate a hydrocarbon membrane that has low cost and high ion selectivity but it turned out that the VO₂+ caused chemical degradation, which led to shortening the battery life drastically.
To develop a membrane with pore sizes smaller than the hydrated size of vanadium ions yet larger than that of the protons, a research team co-led by Professor Hee-Tae Jung and Professor Hee-Tak Kim from the Department of Chemical and Biomolecular Engineering implemented a graphene-oxide framework (GOF) membrane by cross-linking graphene oxide nanosheets. They believed that GOF, having strong ion selectivity, would be a good candidate for the membrane component for the VRFB. The interlayer spacing between the GO sheets limited moisture expansion and provided selective ion permeation.
The GOF membrane increased the capacity retention of the VRFB, which showed a 15 times higher rate than that of perfluorinated membranes. Its cycling stability was also enhanced up to five times, compared to conventional hydrocarbon membranes.
These pore-sized-tuned graphene oxide frameworks will allow pore-sized tuning of membranes and will be applicable to electrochemical systems that utilize ions of various sizes, such as rechargeable batteries and sensors.
Professor Kim said, “Developing a membrane that prevents the mixing of positive and negative active materials has been a chronic issue in the field of redox-flow batteries. Through this research, we showed that nanotechnology can prevent this crossover issue and membrane degradation. I believe that this technology can be applied to various rechargeable batteries requiring large-scale storage.”
This research was published in Nano Letters on May 3.
Figure 1. Electrochemical performances of the VRFBs with Nafion 115, SPAES (sulfonated poly), and GOF/SPAES: discharge capacity
Figure 2. Schematic of the selective ion transfer of hydrated vanadium ions and protons in the GOF membrane and the molecular structure of the GOF membrane, showing that the GO nanosheets are cross-linked with EDA (ethylenediamine)
2018.09.20 View 7569 -
Effective Drug Delivery to Heart with Tannic Acid
(Professor Haeshin Lee from the Department of Chemistry) Typical methods of drug delivery to the heart require surgical procedures involving incisions in the chest wall and bones. To efficiently treat cardiovascular and related vascular diseases without surgery, a KAIST research team developed a heart-targeting drug delivery technology using tannin acid via intravenous systemic injection. This method can be applied to the development of a variety of new protein-based drugs.
Cardiovascular-circulatory disease is currently the second leading cause of death in Korea. A typical example of this disease is myocardial infarction caused by poor oxygen and nutrient supply due to narrowed coronary arteries and poor blood flow to the heart.
Although there have been numerous research projects to develop chemotherapeutic drugs and therapeutic proteins, clinics still rely on surgical procedures. Drug delivery can be an alternative, but it is quite challenging because ceaseless dynamic cycles of the heart and massive exchanges of blood mean administered therapeutics do not stay inside the heart very long.
Professor Haeshin Lee from the Department of Chemistry and his team employed tannic acid (TA), which is known for giving bitter taste to wines. It is one of the most abundant polyphenols and can be easily found in plants, such as fruits, vegetables, cacao, and others. TA has also been used as a multifunctional coating molecule.
Using these properties of TA, the team complexed protein and peptide therapeutics with tannic acid and succeeded in targeting protein and peptide therapeutics to the heart. TA, coated on the surface of a granulated protein complex, helps maintain cardiac function because it adheres to extracellular matrices, elastin, and collagens in heart tissues allowing the protein to stay attached to the heart tissue for a longer period.
The team confirmed that these Tannic-acid-modified proteins stay in blood vessels five days longer than with protein-only injections. Additionally they found that TA-protein complexes do not show any cardiac toxicity and do not cause noticeable pathology.
The team has been continuously developing biomaterials for medical applications by testing various polyphenolic materials that feature adhesive and coating properties, including tannic acid. They have injected a mixture of TA and fibroblast growth factors (FGF) into animal models with myocardial infarctions. After four weeks, they confirmed that the infarction was reduced and the left ventricular pressure and cardiac output were almost normalized.
Professor Lee said, “Although there have been numerous drugs related to heart disease, so far there has not been efficient drug delivery to the heart so this technology will be able to reformulate existing drugs into new and more efficient drugs.”
This research, jointly led by Dr. Ki-Suk Kim from the Predictive Model Research Center, was published in Nature Biomedical Engineering on April 30 ( http://www.nature.com/articles/s41551-018-0227-9 ).
Figure 1. Schematic for the heart-targeting mechanism of TANNylated protein nanocomplexes: (1) size-dependent permeation, (2) phenolic (that is, TA), and (3) internalization by internalization by myoblasts
Figure 2. Effect of TA based protein complexes on cardiac cell transport efficiency and viral gene expression efficiency and therapeutic function in animal models with myocardial infarction
2018.09.18 View 5571 -
Engineered E. coli Using Formic Acid and CO2 As a C1-Refinery Platform Strain
(Figure: Formic acid and CO2 assimilation pathways consisting of the reconstructed THF cycle and reverse glycine cleavage reaction. This schematic diagram shows the formic acid and CO2 assimilation procedure through the pathway. Plasmids used in this study and the genetic engineering performed in this study are illustrated.)
A research group at KAIST has developed an engineered E. coli strain that converts formic acid and CO2 to pyruvate and produces cellular energy from formic acid through reconstructed one-carbon pathways. The strategy described in this study provides a new platform for producing value-added chemicals from one-carbon sources.
Formic acid is a carboxylic acid composed of one carbon. Formic acid was produced from CO2 by the chemical method. Recently, the C1 Gas Refinery R&D Center has successfully developed a biological process that produces formic acid from carbon monoxide for the first time. Formic acid is in a liquid state when at room temperature and atmospheric pressure. In addition, it is chemically stable and less toxic, thus, easy to store and transport. Therefore, it can be used as an alternative carbon source in the microbial fermentation process. In order to produce value-added chemicals using formic acid, a metabolic pathway that converts formic acid into cellular molecules composed of multiple carbons is required. However, a metabolic pathway that can efficiently convert formic acid into cellular molecules has not been developed. This acted as an obstacle for the production of value-added chemicals using formic acid
A research group of Ph.D. student Junho Bang and Distinguished Professor Sang Yup Lee of the Department of Chemical and Biomolecular Engineering addressed this issue. This study, entitled “Assimilation of Formic Acid and CO2 by Engineered Escherichia coli Equipped with Reconstructed One-Carbon Assimilation Pathways”, has been published online in the Proceedings of the National Academy of Sciences of the United States of America (PNAS) on September 18.
There has been increasing interest in utilizing formic acid as an alternative carbon source for the production of value-added chemicals. This research reports the development of an engineered E. coli strain that can convert formic acid and CO2 to pyruvate and produce cellular energy from formic acid through the reconstructed one-carbon pathways.
The metabolic pathway that efficiently converts formic acid and CO2 into pyruvate was constructed by the combined use of the tetrahydrofolate cycle and reverse glycine cleavage reaction. The tetrahydrofolate cycle was reconstructed by utilizing Methylobacterium extorquens formate-THF ligase, methenyl-THF cyclohydrolase, and methylene-THF dehydrogenase. The glycine cleavage reaction was reversed by knocking out the repressor gene (gcvR) and overexpressing the gcvTHP genes that encode enzymes related with the glycine cleavage reaction. Formic acid and CO2 conversion to pyruvate was increased via metabolic engineering of the E. coli strain equipped with the one-carbon assimilation pathway.
In addition, in order to reduce glucose consumption and increase formic acid consumption, Candida boidnii formate dehydrogenase was additionally introduced to construct a cellular energy producing pathway from formic acid. This reduces glucose consumption and increases formic acid consumption.
The reconstructed one-carbon pathways can supply cellular molecules and cellular energies from the formic acid and CO2. Thus, the engineered E. coli strain equipped with the formic acid and CO2 assimilation pathway and cellular energy producing pathway from formic acid showed cell growth from formic acid and CO2 without glucose. Cell growth was monitored and 13C isotope analysis was performed to confirm E. coli growth from the formic acid and CO2. It was found that the engineered E. coli strain sustained cell growth from the formic acid and CO2 without glucose.
Professor Lee said, “To construct the C1-refinery system, a platform strain that can convert one-carbon materials to higher carbon materials needs to be developed. In this report, a one-carbon pathway that can efficiently convert formic acid and CO2 to pyruvate was developed and a cellular energy producing pathway from formic acid was introduced. This resulted in an engineered E. coli strain that can efficiently utilize formic acid as a carbon source while glucose consumption was reduced. The reconstructed one-carbon pathways in this research will be useful for the construction of the C1-refinery system.”
This work was supported by the C1 Gas Refinery Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science and ICT (NRF-2016M3D3A1A01913250). For further information: Sang Yup Lee, Distinguished Professor of Chemical and Biomolecular Engineering, KAIST (leesy@kaist.ac.kr, Tel: +82-42-350-3930)
2018.09.18 View 7013 -
Transfering Nanowires onto a Flexible Substrate
(from left: PhD Min-Ho Seo and Professor Jun-Bo Yoon)
Boasting excellent physical and chemical properties, nanowires (NWs) are suitable for fabricating flexible electronics; therefore, technology to transfer well-aligned wires plays a crucial role in enhancing performance of the devices. A KAIST research team succeeded in developing NW-transfer technology that is expected to enhance the existing chemical reaction-based NW fabrication technology that has this far showed low performance in applicability and productivity.
NWs, one of the most well-known nanomaterials, have the structural advantage of being small and lightweight. Hence, NW-transfer technology has drawn attention because it can fabricate high-performance, flexible nanodevices with high simplicity and throughput.
A conventional nanowire-fabrication method generally has an irregularity issue since it mixes chemically synthesized nanowires in a solution and randomly distributes the NWs onto flexible substrates. Hence, numerous nanofabrication processes have emerged, and one of them is master-mold-based, which enables the fabrication of highly ordered NW arrays embedded onto substrates in a simple and cost-effective manner, but its employment is limited to only some materials because of its chemistry-based NW-transfer mechanism, which is complex and time consuming. For the successful transfer, it requires that adequate chemicals controlling the chemical interfacial adhesion between the master mold, NWs, and flexible substrate be present.
Here, Professor Jun-Bo Yoon and his team from the School of Electrical Engineering introduced a material-independent mechanical-interlocking-based nanowire-transfer (MINT) method to fabricate ultralong and fully aligned NWs on a large flexible substrate in a highly robust manner.
This method involves sequentially forming a nanosacrificial layer and NWs on a nanograting substrate that becomes the master mold for the transfer, then weakening the structure of the nanosacrificial layer through a dry etching process. The nanosacrificial layer very weakly holds the nanowires on the master mold. Therefore, when using a flexible substrate material, the nanowires are very easily transferred from the master mold to the substrate, just like a piece of tape lifting dust off a carpet.
This technology uses common physical vapor deposition and does not rely on NW materials, making it easy to fabricate NWs onto the flexible substrates.
Using this technology, the team was able to fabricate a variety of metal and metal-oxide NWs, including gold, platinum, and copper – all perfectly aligned on a flexible substrate. They also confirmed that it can be applied to creating stable and applicable devices in everyday life by successfully applying it to flexible heaters and gas sensors.
PhD Min-Ho Seo who led this research said, “We have successfully aligned various metals and semiconductor NWs with excellent physical properties onto flexible substrates and applied them to fabricated devices. As a platform-technology, it will contribute to developing high-performing and stable electronic devices.”
This research was published in ACS Nano on May 24.
Figure 1. Photograph of the fabricated wafer-scale fully aligned and ultralong Au nanowire array on a flexible substrate
2018.09.17 View 6382