A5086769474- Microbial Metabolic Engineering and Bioproduction
- Fungal and yeast genetics research
- Gene Regulatory Network Analysis
- CRISPR and Genetic Engineering
- Advanced Fluorescence Microscopy Techniques
- Evolution and Genetic Dynamics
- Biofuel production and bioconversion
- Viral Infectious Diseases and Gene Expression in Insects
- Bacterial Genetics and Biotechnology
- Photosynthetic Processes and Mechanisms
- Monoclonal and Polyclonal Antibodies Research
- Transgenic Plants and Applications
- Click Chemistry and Applications
- Biotin and Related Studies
- DNA Repair Mechanisms
- Enzyme Catalysis and Immobilization
- Advanced Biosensing Techniques and Applications
- Single-cell and spatial transcriptomics
- Plant biochemistry and biosynthesis
- Food Quality and Safety Studies
- Algal biology and biofuel production
- RNA and protein synthesis mechanisms
- Advanced biosensing and bioanalysis techniques
- Bioinformatics and Genomic Networks
- Cell Image Analysis Techniques
Kobe University
2016-2024
Bioprocess Control (Sweden)
2023
Toyota Central Research and Development Laboratories (Japan)
2011-2016
Toyota Motor Corporation (Switzerland)
2013
Osaka University
2003-2011
Japan Science and Technology Agency
2006-2010
The University of Tokyo
2003
Saitama University
1998-2000
Urawa University
1999-2000
The terminator regions of eukaryotes encode functional elements in the 3' untranslated region (3'-UTR) that influence 3'-end processing mRNA, mRNA stability, and translational efficiency, which can modulate protein production. However, contribution these to gene expression remains unclear, therefore their utilization metabolic engineering or synthetic genetic circuits has been limited. Here, we comprehensively evaluated activity 5302 from a total 5880 genes budding yeast Saccharomyces...
A general relationship between fluctuation and response in a biological system is presented. The given by the variance of some quantity, whereas as average change that quantity for parameter change. We propose where two are proportional, similar way to fluctuation–dissipation theorem physics. By studying an evolution experiment fluorescence protein bacteria increases, we confirm our relation observing positive correlation speed phenotypic over clone bacteria. generality well its relevance discussed.
Recombinant yeast strains displaying aheterologous cellulolytic enzymes on their cell surfaces using a glycosylphosphatidylinositol (GPI) anchoring system are promising strategy for bioethanol production from lignocellulosic materials. A crucial step wall localization of the is intracellular transport proteins in cells. Therefore, addition highly efficient secretion signal sequence important to increase amount surface. In this study, we demonstrated effectiveness novel peptide (SP) derived...
Bioactive plant-based secondary metabolites such as stilbenoids, flavonoids, and benzylisoquinoline alkaloids (BIAs) are produced from l-tyrosine (l-Tyr) have a wide variety of commercial applications. Therefore, building microorganism with high l-Tyr productivity (l-Tyr chassis) is immense value for large-scale production various aromatic compounds. The aim this study was to develop an chassis in the nonconventional yeast Pichia pastoris (Komagataella phaffii) produce (resveratrol,...
In the yeast Saccharomyces cerevisiae, terminator sequences not only terminate transcription but also affect expression levels of protein-encoded upstream terminator. The non-conventional Pichia pastoris (syn. Komagataella phaffii) has frequently been used as a platform for metabolic engineering knowledge regarding P. terminators is limited. To explore available to tune protein in pastoris, we created 'terminator catalog' by testing 72 sequences, including from S. cerevisiae or and synthetic...
Secretory production of recombinant proteins is the most important application methylotrophic yeast Pichia pastoris , a species that permits mass heterologous proteins. To date, genetic engineering P. has relied largely on integrative vectors due to lack user-friendly tools. Autonomously replicating plasmids are expected facilitate manipulation; however, existing systems, which use autonomously sequences (ARSs) such as -specific ARS ( PARS1 ), known be inherently unstable for plasmid...
Report28 April 2009Open Access How selection affects phenotypic fluctuation Yoichiro Ito Department of Bioinformatic Engineering, Graduate School Information Science and Technology, Osaka University, Suita, Osaka, Japan ERATO, JST, Search for more papers by this author Hitoshi Toyota Biotechnology, Kunihiko Kaneko Pure Applied Science, University Tokyo, Meguro-ku, Tetsuya Yomo Corresponding Author Frontier Bioscience, Ito1,2, Toyota3, Kaneko2,4 1,2,5 1Department 2ERATO, 3Department...
We aimed at constructing thermostable cellulase variants of cellobiohydrolase II, derived from the mesophilic fungus Phanerochaete chrysosporium, by using an advanced evolutionary molecular engineering method. By aligning amino acid sequences catalytic domains five thermophilic fungal CBH2 and PcCBH2 proteins, we identified 45 positions where genes differ consensus sequence two to CBH2s. with mutations were obtained a cell-free translation system that was chosen for easy evaluation...
Abstract Post-transcriptional upregulation is an effective way to increase the expression of transgenes and thus maximize yields target chemicals from metabolically engineered organisms. Refractory elements in 3′ untranslated region (UTR) that mRNA half-life might be available. In Saccharomyces cerevisiae , several terminator regions have shown activity increasing production proteins by upstream coding genes; among these terminators DIT1 has highest activity. Here, we found two resident...
Summary In our previous study, we serendipitously discovered that protein secretion in the methylotrophic yeast Pichia pastoris is enhanced by a mutation (V50A) mating factor alpha (MFα) prepro‐leader signal derived from Saccharomyces cerevisiae . present investigated 20 single‐amino‐acid substitutions, including V50A, located within MFα peptide, indicating V50A and several single mutations alone provided significant increase production of secreted proteins. addition to hydrophobicity index...
The non-conventional yeast Komagataella phaffii, formerly Pichia pastoris, is a popular host for recombinant protein production. relatively lower gene targeting efficiency observed in this species occurs due to high levels of non-homologous recombination activity. In the current study, we explored function K. phaffii homolog DNA ligase IV (Dnl4p) by creating DNL4-disrupted strain. To assess roles end joining (NHEJ)-related proteins species, strains deleted either or both genes encoding Dnl4p...
Expression of secreted recombinant proteins burdens the protein secretion machinery, limiting production. Here, we describe an approach to improving production by non-conventional yeast Komagataella phaffii comprised genome-wide screening for effective gene disruptions, combining them in a single strain, and recovering growth reduction adaptive evolution. For screen, designed multiwell-formatted, streamlined workflow high-throughput assay single-chain small antibody, which is cumbersome...
Abstract Inducible promoters are essential for precise control of target gene expression in synthetic biological systems. However, engineering eukaryotic is often more challenging than prokaryotic due to their greater mechanistic complexity. In this study, we describe a simple and reliable approach constructing strongly inducible with minimum leakiness yeasts. The results indicate that the yeast-inducible primarily result cryptic transcriptional activation heterologous sequences may be...
Population adaptation can be determined by stochastic switching in living cells. To examine how contributes to the fate decision for a population under severe stress, we constructed an Escherichia coli strain crucially dependent on expression of rewired gene. The gene essential tryptophan biosynthesis, trpC, was removed from native regulatory unit, Trp operon, and placed extraneous control lactose utilisation network. Bistability network provided cells two discrete phenotypes: induced...
The methylotrophic yeast species Komagataella phaffii (synonym: Pichia pastoris) is widely used as a host for recombinant protein production. Although several genetic engineering techniques are being employed on K. phaffii, advanced methods such in vivo DNA assembly this required synthetic biology applications. In study, we established technique accomplishing one-step of multiple fragments and genomic integration phaffii. To concurrently achieve an accurate high-efficient into the target...
<title>Abstract</title> Increasing demand for biologics necessitates the rapid production of antibodies and their mimetics. Herein, we established a workflow to develop fermentation process high-titer small using yeast <italic>Komagataella phaffii</italic>. We screened strains with optimal gene copy number target proteins applied them feedback-controlled, high-cell density fed-batch fermentation, achieved titers 3–7 g/L.