3^rd International Conference on Systems and Synthetic Biology July 20-21, 2017 Munich, Germany

Biography:

Malathi Raman has been the European Cloning & Protein Product Manager at Takara Bio Europe since 2011, and manages Takara’s entire Cloning and Protein product range including the innovative In-Fusion® HD Cloning and Capturem™ Protein and Antibody Purification technologies. Prior to joining Takara, she worked as a Post-Doctoral Research Fellow for 3 years, within the group of Professor Terry Rabbitts, at the LIMM in Leeds, United Kingdom, identifying novel protein-protein interactions involved in the pathogenesis of prostate cancer and Ewing’s sarcoma. She obtained her PhD in Cardiac Genetics from Imperial College London, United Kingdom, in 2008.

Abstract:

There is a constant need for faster and more efficient cloning and protein purification methods at all scales within the Synthetic Biology field. Scientists can accelerate the generation of synthetic gene expression constructs using our innovative Next-Gen In-Fusion^® HD Cloning Plus technology which is fast (15 mins), highly efficient (>95% cloning efficiency), sequence independent (any insert can be cloned into any vector at any locus), seamless (no extra bp), directional, and HTP ready. After generation of the required expression construct and downstream expression of the target protein, Takara offers Next-Gen Capturem™ technology to allow fast (5-15 mins) and easy resin free purification of high quality and concentrated His-tagged proteins or native untagged antibodies. Our revolutionary Capturem technology, available in miniprep, maxiprep and 96-well plate formats, consists of spin columns or plates containing high-capacity nylon membranes immobilized with either Ni²⁺ or Protein A, thus allowing His-tagged protein or antibody purification directly from even complex matrices, such as cell supernatants or serum, within minutes. This talk will review several applications of our technologies including HTP antibody cloning, simplified purification of membrane and secreted proteins, fast hybridoma screening, and rapid immunoprecipitation (IP)/co-immunoprecipitation (Co-IP). We will also introduce our new Capturem Trypsin technology that enables 1 min on-column tryptic digests.

Biography:

Sharon Mendel Williams joined Coventry University as a Lecturer in the School of Life Sciences in the year 2014. She has worked as a Post-doctoral Research Fellow in both departments of Chemistry and Biology, Warwick University. He research focuses on biophysics and biochemistry of proteins, and understanding the mechanisms of enzymes. She has a wide range of experience in molecular biology, biochemistry, and chemistry. She is a member of the Royal Society of Chemistry and has been awarded a grant from the RSC research fund to accomplish her research work.

Abstract:

Lignin is an organic polymer found in the cell walls of plants. Lignin can be used to create biofuels, or as an organic hydrocarbon source for a large variety of chemicals and polymers. However, lignin is very robust and current industrial processes for using it are inefficient. Therefore, a useable biological process for degrading lignin would be of great benefit. Understanding the pathway of lignin degradation, and the byproducts, is essential in order to be able to exploit the use of micro-organisms. Furthermore, we can characterize novel bio-products obtained by enzymatic oxidation of lignin, which could have very interesting applications for industrial biotechnology. In the current project DyP-type peroxidases from Gram-negative Pseudomonas fluorescens Pf-5 and recombinant Sphingobacterium MnSOD1 and MnSOD2 cloned into E. coli and were investigated. These are bacterial enzymes that are already known to degrade lignin. Different genetic mutations were introduced, and the resulting enzymes were characterized by using them with different lignin substrates. The reaction compounds were analyzed by reverse phase HPLC/GC-MS. The goal is to improve the effectiveness of the enzymes, increase the production of the enzymes and degrade the lignin into different and more useable compounds. Any of these goals would be of valuable scientific and commercial benefit.

Biography:

Dr. Iva Falk, PhD. has completed her Ph.D in the field of Medical Technologies. She is working in the Department of Cell Biology and Radiobiology at the Institute of Biophysics of the Czech Academy of Sciences (Brno, CR). She is participating in research that concerns the role of chromatin structure in regulation of cellular processes. Other research interests include DNA damage and repair, carcinogenesis, tumor cells radio-sensitization, and radiobiology.

Abstract:

MDS is heterogeneous group of clonal hematologic disorders characterized by inefficient hematopoiesis. The incidence of MDS is about 4 cases per 100 000 people. The most typical cytogenetic abnormality arising due to still unknown cause and mediated by still unknown mechanism, is a partial or complete deletion of 5q. To address these questions, we isolated lymphocytes and CD34+ hematopoietic cells from healthy donors and MDS patients. By combining 3D-fluorescence in situ hybridization with BAC probes and high-resolution confocal microscopy, we reconstructed higher-order nuclear organization of the CDR (common deleted region) between bands 5q31 and 5q32. Radial and mutual positions of BAC probes, specific for individual chromosomal bands inside the CDR were determined and suggest that higher-order chromatin structure significantly contributes to formation of 5q deletions associated with MDS. Chromatin in the CDR region forms a giant loop that is, by its base, anchored to the nuclear envelope. Though the initial event and the mechanism of the loop base fragmentation has to be further studied, we suppose that close spatial proximity of loci at the loop base, stabilized by anchoring of these loci to the envelope, could simplify deletions of the whole CDR loop.

Biography:

Yaman Arkun research interests are in process dynamics, modeling and control of chemical and biological systems. Research topics include dynamic modeling, model predictive control, large scale complex and hierarchical systems, and optimization. Current application areas are copolymerization reactors, refinery operations, protein folding and systems biology. He completed his BS from Bogazici University, Turkey in 1974 and done his MS in 1976 at University of Minnesota and completed his PhD in 1979 at University of Minnesota. He has held visiting positions at Tennessee Eastman, DuPont and Weyerhauser companies. He has served as the Editor of Journal of Process Control and Associate Editor of Automatica. He has been a trustee and Secretary of Computer Aids in Chemical Engineering (CACHE) Corporation, and a Director of Computers and Systems Technology (CAST), Division of AIChE. He is the recipient of the Donald P Eckman Award given by the American Automatic Control Council. He received TÜBÄ°TAK Science Award in 2004.

Abstract:

Cell signaling is the process by which extracellular information is transmitted into the cell to perform biological functions. RAS-MAPK signaling pathway controls several cellular processes such as cell growth, proliferation, and gene expression. Activation of EGFR (Epidermal growth factor receptor) by binding of its specific ligands starts ERK signaling pathway. Phosphorylated EGFR exhibits great increase in the enzymatic activity of its cytoplasmic tyrosine kinase domain. Adaptor protein Grb2 binds to the phosphorylated RTK followed by recruitment of SOS, forming Grb2-SOS complex. Ras, which is a small GTP binding protein, interacts and transforms to its active conformation by exchanging GDP for GTP. Active Ras acts as an important switch which starts phosphorylation of MAPK pathway that consists of the Raf/MEK/ERK signaling cascade. However, in this paper we present a dynamic model that describes the molecular mechanisms involved in RAS-MAPK signaling. The model is derived from mass-action kinetics and conservation laws. Models exist in the literature for RAS and MAPK pathways separately. Our model combines the two systems and studies their interactions under feedback. Well-known mechanism of deactivation of the Grb2-SOS complex by ERK is included as an inhibitory feedback action in the model. Bistability (i.e., ability to switch between two stable steady-states separated by an unstable steady-state) is a desired trait that most biological processes exhibit. We establish conditions under which bistability and oscillations exist for this important pathway. In particular, we show how the negative and positive feedback loops affect the dynamic characteristics that determine the cellular outcome.

Biography:

Xiaolian Gao is professor of Biology and Biochemistry at University of Houston.  Her major research interests are related to large scale biology. Her lab uses chemical and biophysical methods in combination to address questions of biomolecules concerning how their structures, molecular dynamics, intermolecular interactions, and molecular recognition play roles in biological processes.  In synthetic biology, she has led projects of miniaturized parallel production using digital photochemistry for programmable oligonucleotide synthesis on microchips, which was now advanced to become a revolutionary technology for massive production of oligonucleotides that fulfill the needs of rapid progress of today’s DNA technologies, including gene/genome synthesis, sequencing of the various nucleic acid molecules.  In a recent report, collaborators and students of Dr. Gao has established robust laboratory work flow for accurate multiplex gene synthesis [reported in Science Reports “High-fidelity de novo synthesis of pathways using microchip-synthesized oligonucleotides and general molecular biology equipment” (2017) in print.. Dr.Gao is also a proficient structural biologist for using NMR methods to elucidate structures of ligand-DNA complexes.

Abstract:

This presentation will describe our synthetic biology project aiming a streamlining process of multiplex high fidelity gene synthesis using microchip oligo building blocks. This process features miniaturization, computation bioinformatics design, optimized work flow, low material consumption, long and high sequence accuracy, low error DNA constructs through efficient production process. Specifically, our work established a simple and easy to use flow column method (immobilized cellulose-binding-mutS column) to remove error-containing sequences from the final oligo gene-building blocks which are designed as such that they can be processed by ligation and PCR to give defined long (kb) DNA constructs. The reported workflow required about an hour of bench time for oligo processing, and attained less than 1 error per kb DNA, which is translated to ~80% success rate of full length EGFP (720 bp) gene cloning. The workflow hands more than ten genes in parallel. Has the potential for application in pathway gene cluster synthesis.

Biography:

John B Carrigan has completed his PhD in 2005 from the University College Dublin and carried out postdoctoral both in Dublin and in Copenhagen. He has been involved with several startups, most recently biobased advanced materials company, Cellulac Ltd. He is the CSO in SOSV responsible for scientific due diligence, recruitment, product development analysis in addition to other work. He has published several papers in the area of protein engineering, enzymology, metabolomics and cellulosic biofuels

Abstract:

SOSV’s Cork-based global bioaccelerator initiative, which is dedicated to funding and building startups for the purpose of aiding humanity. Having founded the worlds first life sciences accelerator in Cork in 2014. SOSV now operates two accelerator programs, IndieBio based in San Francisco and RebelBio again based in Cork. These accelerators are responsible or establishing many synthetic biology startups around the world including Perfect Day Foods, Memphis Meats, Microsynbiotix and the German based Saphium Biotech. We provide the mechanism by which young scientists, entrepreneurs and tinkerers can shape their own destiny and make something that matters. RebelBio provides seed funding and mentorship to drive the transition of science to a business in only four months, before launching its graduate companies into the world of biotechnology to make their fortune, buffered by the company’s many alumni, partners and partner investors.

Biography:

Abstract:

Biography:

Ralf Takors completed his PhD in Biochemical Engineering in 1997 and received Habilitation in Metabolic Engineering in 2004, both at Forschungszentrum Jülich GmbH and RWTH Aachen. He has worked at the Evonik Industries till 2009, where he was responsible for bioprocess development, metabolic engineering and systems biology research. In 2009, he became the Head of the Institute of Biochemical Engineering at the University Stuttgart. Research interests are Systems Metabolic Engineering and Synthetic Biology for the development of novel bioprocesses.

Abstract:

Sooner or later novel processes with recombinant producers should find their way from the labs to large-scale fermenters to commercialize the product. However, successful scale-up is often hampered by harsh production conditions which expose the strains to frequently changing substrate supply due to technical limits of mixing. In a series of systems biology experiments, metabolic and transcriptional responses of E. coli to large-scale conditions were studied. Short- and long-term consequences of changing glucose and nitrogen availabilities were investigated. Applying typical mixing times of 110 seconds, more than 600 genes were found to be frequently up- and down regulated. Accordingly, cellular maintenance demands increased by 40 to 50% which was identified by measurements and by systems modeling and which created a list of gene candidates for smart genome reduction. ppGpp, the alarmone of stringent response, turned out to be protagonist of the observed regulation programs. Engineering of key genes of the stringent response together with modulations in central metabolism finally yielded E. coli HGT. The strain shows 10 fold increased glucose uptake rates (compared to the native maintenance demands) under resting or slow growth conditions which are a preferred production scenario. The surplus of glucose uptake is available as pyruvate to enable likewise utilization which is why E. coli HGT represents a novel chassis for the production of pyruvate derived products in large-scale.

Biography:

Betty Lee has a PhD from Dartmouth Medical School, USA and MS in Clinical Chemistry from the University of Windsor, Canada. She has done his second MS Biochemistry from LSU Medical Center, USA. She completed her Post-doctoral training at the National Institutes of Health, USA. She is currently working as a Licensing Officer with the US government. She educates industries and academia about the Export Administration Regulations (EAR) and participates in outreach. In addition, she participated in the policy review of the Executive Order titled “Optimizing the security of biological select agents and toxins in the United States” signed by President Obama on July 2, 2010

Abstract:

Balancing biosecurity and legitimate life science research has become a priority in recent years. The rise of biotechnology and informatics has made rapid advances in the 21^st century. Such a convergence of biology and technology increases the pace of biological findings and the emergence of new technologies. Emerging technologies that could be used for biological warfare poses a formidable challenge because of the unpredictable nature of science. Dual use research in the life sciences requires some oversight by the government and funding agencies. The published results of scientific research could be used to improve health or agricultural products or they could be used to enable bioterrorism. Life Sciences research is conducted increasingly in an interdisciplinary and international environment. Informatics, systems biology, nanotechnology, and synthetic biology are at the forefront of such endeavors. Oversight of such research is essential to continue the free exchange of information and also balance the national security concerns. One of the tools that are at the nexus of biosecurity and life sciences is export control. The session will provide an overview of the biological agents (viruses, bacteria and toxins), genetic elements (DNA, plasmids, vectors) which are currently controlled on the Australia Group Control List and Commerce Control List for exports. The Australia Group is an informal forum of countries which seeks to ensure that exports do not contribute to the development of chemical or biological weapons. Topics of discussion will include deemed exports, fundamental research and technology transfer.

Biography:

Erchin Serpedin is currently a Professor at Texas A&M University in College Station, TX. He is the author of more than 140 journal papers, 250 conference papers, and 4 books. His research interests lie in the areas of computational biology, systems biology, signal processing and machine learning

Abstract:

We propose a novel consistency-classification framework that enables the assessment of consistency and classification performance of a biomarker discovery algorithm. The proposed evaluation protocol is based on random resampling those models the variation in the experiment size. The metagenomic data matrix is modeled as a superposition of two matrices. The first matrix is a low-rank matrix that depicts the abundance levels of the irrelevant bacteria. The second matrix is a sparse matrix that describes the abundance levels of the bacteria that are differentially abundant between different phenotypes. We propose a novel Robust Principal Component Analysis (RPCA) based biomarker discovery algorithm to recover the sparse matrix. RPCA is a multivariate feature selection approach that processes the features collectively rather than individually. Comprehensive comparisons of RPCA with the state-of-the-art algorithms on two realistic datasets show that RPCA consistently outperforms the existing state-of-the-art algorithms in terms of classification accuracy and reproducibility performance. Thus, the proposed RPCA-based biomarker detection algorithm provides a high reproducibility performance irrespective of the complexity of the dataset and the number of selected biomarkers. RPCA selects also biomarkers with quite high discriminative accuracy. Therefore, RPCA appears to represent a very consistent and accurate methodology for selecting taxonomical biomarkers in microbial populations.

Biography:

Maria A Ragusa has completed her PhD in Cellular and Developmental Biology and Post-doctoral studies from Palermo University. She is an Assistant Professor of Molecular Biology at the University of Palermo (Italy). She has published more than 15 peer reviewed papers in reputed international journals and has been serving as peer reviewer for many international journals. Her research work comprises gene transcription regulation during embryo development and early molecular defense strategies activated by embryos in response to exposition to stress agents. Recently, she also studied the effects of HPV infection on semen

Abstract:

Echinoderms represent a very fascinating phylum belonging to deuterostome superphylum, including also hemichordates, tunicates and vertebrates. Among echinoderms, sea urchin species are studied worldwide as excellent model organisms for the study of developmental biology, and are suitable as bioindicators. During a study on the defense strategies activated by Paracentrotus lividus sea urchin embryos in response to CdCl₂, we isolated five metallothionein (MT) cDNAs. Two of the five MT genes were constitutively expressed, whereas the other three genes appear to be specifically switched-on in response to cadmium treatment. With the aim of better understanding the evolutionary relationships, functional variety, and the utilization of MTs during development, the gene organization of P. lividus MTs was analyzed and their mRNA expression patterns were unveiled. Particularly, we determined the expression profiles and the spatial patterns of MT transcripts during development and after metal and antimicrobial drug treatments. MTs show a high polymorphism both in sequence/structure and expression pattern. In the light of our findings it seems reasonable to consider MT7 and MT8 as the major variants associated with physiological functions, playing their major roles in metal homeostasis and redox activity in ecto-, meso- and endo-dermal tissues. On the other hand, a metal detoxification role can be attributed to MT4, 5, and 6, particularly important in mesenchyme cells for the skeletogenic pathway. Since differences not only between the two classes occur, each isoform would correspond to a more definite physiological function. For these interesting results we started to study the mechanisms that control MT gene expression in sea urchin embryogenesis.

Biography:

Guido Cimoli completed his Master’s degree in Biology at University of Genoa and worked as a Researcher at National Cancer Institute. He completed his PhD in Molecular Pharmacology at University of Pavia. He worked as a Bio-pharma Application Specialist and Project Leader at Tecan Group Ltd., from February 2004 to December 2007. He is currently responsible for the sales development for automation in Europe and Middle East.

Abstract:

In the last few years, the branch of biology (or of engineering, as some says) called synthetic biology has definitely taken the center of the stage. This oxymoronic branch is quickly spreading from agriculture to medicine, from students to Nobel Prize laureates, from engineers to molecular biologist and at every step increasing sophistication and complexity. The complexity stems both from intrinsic sophistication of the biological circuits that wants to be mimicked or created and also from the large amount of steps that have to be physically performed to transform an in silico circuit to a real, organic one. While the complexity at scientific level can be only tamed by continuous development of knowledge (and its spreading) and the development of more powerful software and deeper databases, the complexity at practical level can be tremendously decreased by the introduction of automation procedures. Automating tasks which requires high manual skills while being of limited intellectual complexity (think of separating interphases), or tasks that requires extremely high precision and management of ultra-low volumes (think of low volumes qPCR), or highly repetitive ones and at high frequency (think of colony picking or screening) can only improve the outcome with respect to manual performance. Automation does not only improve the quality of the data esults of a given task but can also bring other advantage like freeing skilled personnel for more important tasks, reduce burden of labor, scale up processes, standardize operations within the laboratory, examples will be given in the presentation.

Biography:

Lenka Dohnalová has completed her BSc from University of Chemical Technology Prague, where she is currently completing her Master’s studies. She spent 1.5 years as a visiting student in the lab of Eran Elinav at the Weizmann Institute of Science, where she investigated the role of the intestinal microbiome in metabolism and immunity.

Abstract:

Despite the effectiveness of commonly used dietary weight reduction strategies, the majority of dieters fail to maintain the reduced weight in the long run. Recently, we uncovered a contribution of the intestinal microbiome to post-dieting weight regain by flavonoid-mediated regulation of energy expenditure. We identified a stable microbial composition that persists after obesity and promotes faster weight regain after high-fat diet (HFD) re-exposure. Furthermore, we demonstrated that fecal microbiome transplantation as well as flavonoid supplementation ameliorates the secondary weight gain. Additionally, we developed a machine leasing-based algorithm based on 16S rRNA data for the accurate prediction of weight-regain. Our data therefore suggest a possible implication of microbiome and metabolome in the personalized prediction and treatment of post-dieting weight regain.

Biography:

Qiang Yan is a PhD candidate working under the supervision of Dr. Stephen S Fong in the Department of Chemical and Life Science Engineering, Virginia Commonwealth University (VCU). His research interest focused on using molecular genetic techniques, synthetic biology and computational models to design modify and improve organism as workhorse to solve real-world problems.

Abstract:

Annually, large amounts of waste material are generated from seafood (crab, shrimp and lobster shells) that typically go to landfills with high expense (e.g. $150/ton in Australia). The biopolymer chitin makes up ~40% total weight of such waste and it can potentially be used as a sustainable feedstock for value-added products. In this study, we developed a consolidated bioprocess to produce N-acetylneuraminic acid (NeuNAc) directly from chitin using the chitinolytic organism Serratia marcescens by developing and implementing genetic engineering tools to modify S. marcescens. Starting from RNA sequencing data, 10 native promoters of varying strength were identified and characterized using a fluorescent reporter gene (sfGFP). Two heterologous genes (N-acetylglucosamine isomerase and NeuNAc lyase) were introduced in S. marcescens, and expression was transcriptionally controlled using three different strength promoters (high-high, medium-medium, and low-low). An initial 34.8 mg/L NeuNAc was produced under a high-high promoter combination. Improvements to NeuNAc were conducted by changing expression strength of the two heterologous genes to balance metabolic flux. Using a high strength promoter for the isomerase and a medium strength promoter for the lyase improved both NeuNAc production (1.42-fold) and growth (2.57-fold). Further improvements were tested by characterizing the kinetics of each heterologous enzyme and implementing alternative genes. Swapping N-acetylglucosamine isomerase gene did not show any improvement in production; however, swapping the second-step reversible NeuNAc lyase (nanA) to an irreversible NeuNAc synthase (neuB) improved NeuNAc production (3.25-fold). Overall, we identified and characterized promoter sequences that can be used for genetic engineering of S. marcescens and have implemented the tools to demonstrate and improve production of NeuNAc.

Biography:

Albert Jeltsch finished his PhD on Restriction Endonucleases at University Hannover in 1994. Afterwards, he started working on DNA Methyltransferases at Justus-Liebig University Giessen and at the Jacobs University Bremen. Since 2011, he is a Professor of Biochemistry at the University Stuttgart. He is the recipient of the Gerhard-Hess award (DFG) and BioFuture award (BMBF). He has long standing expertise in the biochemical study of DNA and protein methyltransferases, methyl lysine reading domains and in rational and evolutionary protein design. His work has been published in >200 publications in peer reviewed journals and is in the editorial boards of several journals

Abstract:

Investigation of the fundamental role of epigenetic processes depends on the development of methods, which enable live-cell detection of epigenetic modifications with locus-specific resolution. Here, we address this urgent technological demand by developing four modular fluorescence complementation-based epigenetic biosensors for live cell microscopy applications. In these tools, we combine the high DNA sequence specificity of engineered anchor proteins, with the great versatility of chromatin reading domains as natural detector modules for the recognition of target epigenetic marks. Simultaneous readout of DNA sequence and epigenetic mark is detected as reconstituted fluorescent signals that arise upon binding of the anchor and detectors modules in close spatial proximity, within the nuclei of living cells. With this approach we could directly detect DNA methylation and histone 3 lysine 9 trimethylation at defined, endogenous genomic sites, in several mouse and human cell lines. Furthermore, we could follow dynamic changes in these marks with locus-specific resolution upon drug treatment or induction of epigenetic enzymes. We anticipate that this versatile technology will play an important role in improving our understanding of how specific epigenetic signatures are set, erased and maintained during embryonic development or the onset of diseases.

Biography:

Yuquan Xu has completed his PhD from China Agricultural University and postdoctoral studies from University of Arizona and University of California, San Diego. His research fouses on combinatorial biosynthesis of fungal polyketides. He has published more than 23 papers in reputed journals.

Abstract:

Combinatorial biosynthesis aspires to exploit the incorporation of different microbial anabolic pathways to engineer the synthesis of new chemical entities. The modular enzymes composed by functionally independent domains, e.g., polyketide synthases (PKS), are suitable test case for the modularization of secondary metabolic pathways into “build–couple–pair” synthetic schemes. Fungal benzenediol lactones (BDLs) polyketides provide an opportunity for combinatorial biosynthesis. Fungal BDLs are important pharmacophores with wide-ranging bioactivities, and their biosynthesis involves a pair of collaborating iterative polyketide synthases (iPKSs): A highly reducing iPKS (hrPKS) with product that is further elaborated by a non-reducing iPKS (nrPKS) to yield a 1,3-benzenediol moiety bridged by a macrolactone. Co-expressing random heterocombinations of hrPKSs and nrPKSs from different BDL biosynthetic pathways in Saccharomyces cerevisiae lead to the one-pot, one-step combinatorial biosynthesis of structurally diverse polyketides. In addition, the chemical diversity was further increased using heterologous tailoring enzymes such as glycosyltransferase and methyltransferase. Through heterologous expression and domain recombination to create hybrid enzymes, the product template (PT) domains in fungal nrPKSs that catalyze the first-ring cyclization of the benzenediol moiety can be used heterologously to create unnatural products with different polyketide folding modes. This folding mode difference can be programed by reshaping the cyclization chamber of a PT domain by only three selected point mutations. In addition, unnatural products can be generated via shuffling the nrPKS subunit if carefully tune the selectivity of the starter unit (SAT) and the TE domain. Our work provides a biosynthetic tool to generate unnatural polyketides as an unexplored source of chemical diversity and novelty, ready to be exploited for drug discovery, and these results demonstrated the potentials of combinatorial biosynthesis to produce new product with structure variety when considering the rules of enzyme activity and selectivity, shedding lights to further engineer the metabolic pathways in fungi.