A5039092199- Studies on Chitinases and Chitosanases
- Enzyme Production and Characterization
- bioluminescence and chemiluminescence research
- RNA and protein synthesis mechanisms
- Photoreceptor and optogenetics research
- Bacterial Genetics and Biotechnology
- Protein Hydrolysis and Bioactive Peptides
- Legume Nitrogen Fixing Symbiosis
- Advanced Fluorescence Microscopy Techniques
- Advanced biosensing and bioanalysis techniques
- Photochromic and Fluorescence Chemistry
- Protein Structure and Dynamics
- CRISPR and Genetic Engineering
- Virus-based gene therapy research
- Insect symbiosis and bacterial influences
- Light effects on plants
- Synthesis of Indole Derivatives
- CAR-T cell therapy research
- Supercapacitor Materials and Fabrication
- Advancements in Battery Materials
- Microbial Metabolism and Applications
- Advanced Memory and Neural Computing
- Advanced Biosensing Techniques and Applications
- Invertebrate Immune Response Mechanisms
- Pineapple and bromelain studies
Yale University
2022-2025
Niigata University
2008-2024
Center for Systems Biology
2022-2023
Yale New Haven Health System
2023
Yale Cancer Center
2023
The University of Tokyo
2019-2022
Tokyo University of the Arts
2019-2022
Osaka University
2016-2019
Niigata Institute of Technology
2016-2018
Pennsylvania State University
2002-2016
Abstract Luminescence imaging has gained attention as a promising bio-imaging modality in situations where fluorescence cannot be applied. However, wider application to multicolour and dynamic is limited by the lack of bright luminescent proteins with emissions across visible spectrum. Here we report five new spectral variants protein, enhanced Nano-lantern (eNL), made concatenation brightest luciferase, NanoLuc, various colour hues fluorescent proteins. eNLs allow five-colour live-cell...
Summary The carbon storage regulatory system of Escherichia coli controls the expression genes involved in carbohydrate metabolism and cell motility. CsrA binding to glgCAP transcripts inhibits glycogen by promoting mRNA decay. CsrB RNA functions as an antagonist sequestering this protein preventing its action. In paper, we elucidate further mechanism CsrA‐mediated glgC regulation. Results from gel shift assays demonstrate that several molecules can bind each transcript. footprinting studies...
A chitinase gene of Bacillus circulans WL-12 was cloned into Escherichia coli by transforming HB101 cells with a recombinant plasmid composed chromosomal DNA fragments prepared from B. and the vector pKK223-3. sequencing analysis revealed that region necessary for normal expression activity contained one open reading frame 2097 base pairs which codes precursor A1. The A1 long signal sequence 41 amino acids an extremely N-terminal hydrophilic segment 15 acids. Cloned produced in E. had at its...
Bacillus circulans WL-12, isolated as a yeast cell wall-lytic bacterium, secretes variety of polysaccharide-degrading enzymes into culture medium. When chitinases the bacterium were induced with chitin, six distinct chitinase molecules detected in supernatant. These (A1, A2, B1, B2, C, and D) showed following sizes isoelectric points: Mr 74,000, pI 4.7 (A1); 69,000, 4.5 (A2); 38,000, 6.6 (B1); 5.9 (B2); 39,000, 8.5 (C); 52,000, 5.2 (D). Among these chitinases, A1 A2 had highest...
To discover the individual roles of chitinases from Serratia marcescens 2170, A, B, and C1 (ChiA, ChiB, ChiC1) were produced by Escherichia coli their enzymatic properties as well synergistic effect on chitin degradation studied. All three showed a broad pH optimum maintained significant chitinolytic activity between 4 10. ChiA was most active enzyme toward insoluble chitins, but ChiC1 soluble derivatives among chitinases. Although all released (GlcNAc)2 almost exclusively colloidal chitin,...
The third chitinase gene (chiC) of Serratia marcescens 2170, specifying chitinases C1 and C2, was identified. Chitinase lacks a signal sequence consists catalytic domain belonging to glycoside hydrolase family 18, fibronectin type III-like (Fn3 domain) C-terminal chitin-binding (ChBD). C2 corresponds the is probably generated by proteolytic removal Fn3 ChBDs. loss portion reduced hydrolytic activity towards powdered chitin regenerated chitin, but not colloidal glycol illustrating importance...
To carry out a genetic analysis of the degradation and utilization chitin by Serratia marcescens 2170, various Tn5 insertion mutants with characteristic defects in chitinase production were isolated partially characterized. Prior to isolation mutants, proteins secreted into culture medium presence analyzed. Four chitinases, A, B, C1, C2, among other proteins, detected supernatant S. 2170. All four chitinases 21-kDa protein (CBP21) lacking activity showed binding activity. Cloning sequencing...
The gene (chiD) encoding the precursor of chitinase D was found to be located immediately upstream chiA gene, A1, which is a key enzyme in system Bacillus circulans WL-12. Sequencing analysis revealed that deduced polypeptide encoded by chiD 488 amino acids long and distance between coding regions genes 103 bp. Remarkable similarity observed N-terminal one-third C-terminal A1. 47-amino-acid segment (named ND) showed 61.7% acid match with (CA) following 95-amino-acid (R-D) 62.8 60.6% matches,...
ABSTRACT The global regulator CsrA (carbon storage regulator) of Escherichia coli is a small RNA binding protein that represses various metabolic pathways and processes are induced in the stationary phase growth, while it activates certain exponential functions. Both repression activation by involve posttranscriptional mechanisms, which to mRNA leads decreased or increased transcript stability, respectively. also binds untranslated RNA, CsrB, forming ribonucleoprotein complex, antagonizes...
We report development of the first genetically encoded bioluminescent indicator for membrane voltage called LOTUS-V. Since it is bioluminescent, imaging LOTUS-V does not require external light illumination. This allows bidirectional optogenetic control cellular activity triggered by Channelrhodopsin2 and Halorhodopsin during imaging. The other advantage robustness a signal-to-background ratio (SBR) wherever expressed, even in specimens where autofluorescence from environment severely...
The pleiotropic effects of human disease and the complex nature gene-interaction networks require knock-in mice allowing for multiplexed gene perturbations. Here we describe a series with C57BL/6 background conditional or constitutive expression LbCas12a high-fidelity enhanced AsCas12a, which were inserted at Rosa26 locus. Cas12a in did not lead to discernible pathology enabled efficient genome engineering. We used retrovirus-based immune-cell engineering CD4+ CD8+ T cells, B cells...
The widely conserved protein CsrA (carbon storage regulator A) globally regulates bacterial gene expression at the post-transcriptional level. In many species, activity is governed by untranslated sRNAs, CsrB and CsrC in Escherichia coli, which bind to multiple dimers, sequestering them from lower affinity mRNA targets. Both synthesis turnover of CsrB/C are regulated. Their requires housekeeping endonuclease RNase E activated presence a preferred carbon source via binding EIIAGlc glucose...
Because cyanobacteriochrome photoreceptors need only a single compact domain for chromophore incorporation and absorption of visible spectra including the long-wavelength far-red region, these molecules have been paid much attention application to bioimaging optogenetics. Most cyanobacteriochromes, however, drawback incorporate phycocyanobilin that is not available in mammalian cells. In this study, we focused on biliverdin (BV) intrinsic absorbs region revealed replacement four residues was...
Fluorescent and bioluminescent genetically encoded Ca2+ indicators (GECIs) are an indispensable tool for monitoring dynamics in numerous cellular events. Although fluorescent GECIs have a high spatiotemporal resolution, their application is often confined to short-term imaging due the external illumination that causes phototoxicity autofluorescence from specimens. Bioluminescent overcome these pitfalls with enhanced compatibility optogenetic manipulation photophysiological processes;...
Three new photochromic coumarins were synthesized. Fluorescence of the open form 7-hydroxy-3,4-bisthiazolyl-coumarin increased to 1400% by changing pH only slightly from 6.05 7.58. This was subsequently quenched 1.5% maximum intensity at UV photostationary state in water-methanol media.
The rice class I chitinase OsChia1b, also referred to as RCC2 or Cht-2, is composed of an N-terminal chitin-binding domain (ChBD) and a C-terminal catalytic (CatD), which are connected by proline- threonine-rich linker peptide. Because the ability inhibit fungal growth, OsChia1b gene has been used produce transgenic plants with enhanced disease resistance. As initial step toward elucidating mechanism hydrolytic action antifungal activity, full-length structure was analyzed X-ray...
The third chitinase gene (chiC) of Serratia marcescens 2170, specifying chitinases C1 and C2, was identified. Chitinase lacks a signal sequence consists catalytic domain belonging to glycoside hydrolase family 18, fibronectin type III-like (Fn3 domain) C-terminal chitin-binding (ChBD). C2 corresponds the is probably generated by proteolytic removal Fn3 ChBDs. loss portion reduced hydrolytic activity towards powdered chitin regenerated chitin, but not colloidal glycol illustrating importance...
The sarco/endoplasmic reticulum (SR/ER) is the foremost intercellular Ca2+ store (at submillimolar concentrations), playing a crucial role in controlling intracellular levels. For investigation of SR/ER dynamics cells, fluorescent protein-based genetically encoded calcium indicators (GECIs) with low affinity have been used. Recently, bioluminescent GECIs high brightness reported to counter constraints fluorescence imaging, such as phototoxicity. However, their and limited for imaging...