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1.
Nat Commun ; 14(1): 3365, 2023 Jun 08.
Article in English | MEDLINE | ID: mdl-37291127

ABSTRACT

Spin Seebeck effect (SSE) refers to the generation of an electric voltage transverse to a temperature gradient via a magnon current. SSE offers the potential for efficient thermoelectric devices because the transverse geometry of SSE enables to utilize waste heat from a large-area source by greatly simplifying the device structure. However, SSE suffers from a low thermoelectric conversion efficiency that must be improved for widespread application. Here we show that the SSE substantially enhances by oxidizing a ferromagnet in normal metal/ferromagnet/oxide structures. In W/CoFeB/AlOx structures, voltage-induced interfacial oxidation of CoFeB modifies the SSE, resulting in the enhancement of thermoelectric signal by an order of magnitude. We describe a mechanism for the enhancement that results from a reduced exchange interaction of the oxidized region of ferromagnet, which in turn increases a temperature difference between magnons in the ferromagnet and electrons in the normal metal and/or a gradient of magnon chemical potential in the ferromagnet. Our result will invigorate research for thermoelectric conversion by suggesting a promising way of improving the SSE efficiency.


Subject(s)
Electricity , Electrons , Animals , Estrus , Hot Temperature , Oxides , Oxygen
2.
J Biol Chem ; 299(5): 104652, 2023 05.
Article in English | MEDLINE | ID: mdl-36990220

ABSTRACT

N-formyl methionine (fMet)-containing proteins are produced in bacteria, eukaryotic organelles mitochondria and plastids, and even in cytosol. However, Nα-terminally formylated proteins have been poorly characterized because of the lack of appropriate tools to detect fMet independently of downstream proximal sequences. Using a fMet-Gly-Ser-Gly-Cys peptide as an antigen, we generated a pan-fMet-specific rabbit polyclonal antibody called anti-fMet. The raised anti-fMet recognized universally and sequence context-independently Nt-formylated proteins in bacterial, yeast, and human cells as determined by a peptide spot array, dot blotting, and immunoblotting. We anticipate that the anti-fMet antibody will be broadly used to enable an understanding of the poorly explored functions and mechanisms of Nt-formylated proteins in various organisms.


Subject(s)
Antibodies , Antibody Specificity , N-Formylmethionine , Proteins , Animals , Humans , Rabbits , Antibodies/analysis , Antibodies/immunology , Bacteria/chemistry , Cytosol/metabolism , Immune Sera/analysis , Immune Sera/immunology , Immunoblotting , Mitochondria/metabolism , N-Formylmethionine/analysis , N-Formylmethionine/immunology , Proteins/analysis , Proteins/chemistry , Proteins/immunology , Proteins/metabolism , Saccharomyces cerevisiae/chemistry
3.
Biomedicines ; 10(9)2022 Aug 27.
Article in English | MEDLINE | ID: mdl-36140200

ABSTRACT

Extensive progress in understanding the molecular mechanisms of cancer growth and proliferation has led to the remarkable development of drugs that target cancer-driving molecules. Most target molecules are proteins such as kinases and kinase-associated receptors, which have enzymatic activities needed for the signaling cascades of cells. The small molecule inhibitors for these target molecules greatly improved therapeutic efficacy and lowered the systemic toxicity in cancer therapies. However, long-term and high-dosage treatment of small inhibitors for cancer has produced other obstacles, such as resistance to inhibitors. Among recent approaches to overcoming drug resistance to cancers, targeted protein degradation (TPD) such as proteolysis-targeting chimera (PROTAC) technology adopts a distinct mechanism of action by which a target protein is destroyed through the cellular proteolytic system, such as the ubiquitin-proteasome system or autophagy. Here, we review the currently developed PROTACs as the representative TPD molecules for cancer therapy and the N-degrons of the N-degron pathways as the potential TPD ligands.

4.
Adv Mater ; 34(45): e2203558, 2022 Nov.
Article in English | MEDLINE | ID: mdl-36122902

ABSTRACT

Physical unclonable function (PUFs) utilize inherent random physical variations of solid-state devices and are a core ingredient of hardware security primitives. PUFs promise more robust information security than that provided by the conventional software-based approaches. While silicon- and memristor-based PUFs are advancing, their reliability and scalability require further improvements. These are currently limited by output fluctuations and associated additional peripherals. Here, highly reliable spintronic PUFs that exploit field-free spin-orbit-torque switching in IrMn/CoFeB/Ta/CoFeB structures are demonstrated. It is shown that the stochastic switching polarity of the perpendicular magnetization of the top CoFeB can be achieved by manipulating the exchange bias directions of the bottom IrMn/CoFeB. This serves as an entropy source for the spintronic PUF, which is characterized by high entropy, uniqueness, reconfigurability, and digital output. Furthermore, the device ensures a zero bit-error-rate under repetitive operations and robustness against external magnetic fields, and offers scalable and energy-efficient device implementations.

5.
Mol Plant ; 15(6): 991-1007, 2022 06 06.
Article in English | MEDLINE | ID: mdl-35524409

ABSTRACT

Salicylic acid (SA) plays an important role in plant immune response, including resistance to pathogens and systemic acquired resistance. Two major components, NONEXPRESSOR OF PATHOGENESIS-RELATED GENES (NPRs) and TGACG motif-binding transcription factors (TGAs), are known to mediate SA signaling, which might also be orchestrated by other hormonal and environmental changes. Nevertheless, the molecular and functional interactions between SA signaling components and other cellular signaling pathways remain poorly understood. Here we showed that the steroid plant hormone brassinosteroid (BR) promotes SA responses by inactivating BR-INSENSITIVE 2 (BIN2), which inhibits the redox-sensitive clade I TGAs in Arabidopsis. We found that both BR and the BIN2 inhibitor bikinin synergistically increase SA-mediated physiological responses, such as resistance to Pst DC3000. Our genetic and biochemical analyses indicated that BIN2 functionally interacts with TGA1 and TGA4, but not with other TGAs. We further demonstrated that BIN2 phosphorylates Ser-202 of TGA4, resulting in the suppression of the redox-dependent interaction between TGA4 and NPR1 as well as destabilization of TGA4. Consistently, transgenic Arabidopsis overexpressing TGA4-YFP with a S202A mutation displayed enhanced SA responses compared to the wild-type TGA4-YFP plants. Taken together, these results suggest a novel crosstalk mechanism by which BR signaling coordinates the SA responses mediated by redox-sensitive clade I TGAs.


Subject(s)
Arabidopsis Proteins , Arabidopsis , Arabidopsis/metabolism , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Basic-Leucine Zipper Transcription Factors/metabolism , Brassinosteroids/metabolism , Brassinosteroids/pharmacology , Gene Expression Regulation, Plant , Immunity , Phosphorylation , Protein Kinases/metabolism , Salicylic Acid/metabolism , Salicylic Acid/pharmacology , Transcription Factors/genetics , Transcription Factors/metabolism
6.
Sci Rep ; 11(1): 20884, 2021 Oct 22.
Article in English | MEDLINE | ID: mdl-34686705

ABSTRACT

Electrical conduction in magnetic materials depends on their magnetization configuration, resulting in various magnetoresistances (MRs). The microscopic mechanisms of MR have so far been attributed to either an intrinsic or extrinsic origin, yet the contribution and temperature dependence of either origin has remained elusive due to experimental limitations. In this study, we independently probed the intrinsic and extrinsic contributions to the anisotropic MR (AMR) of a permalloy film at varying temperatures using temperature-variable terahertz time-domain spectroscopy. The AMR induced by the scattering-independent intrinsic origin was observed to be approximately 1.5% at T = 16 K and is virtually independent of temperature. In contrast, the AMR induced by the scattering-dependent extrinsic contribution was approximately 3% at T = 16 K but decreased to 1.5% at T = 155 K, which is the maximum temperature at which the AMR can be resolved using THz measurements. Our results experimentally quantify the temperature-dependent intrinsic and extrinsic contributions to AMR, which can stimulate further theoretical research to aid the fundamental understanding of AMR.

7.
Nano Lett ; 20(11): 7803-7810, 2020 Nov 11.
Article in English | MEDLINE | ID: mdl-33054243

ABSTRACT

Coupling of spin and heat currents enables the spin Nernst effect, the thermal generation of spin currents in nonmagnets that have strong spin-orbit interaction. Analogous to the spin Hall effect that electrically generates spin currents and associated electrical spin-orbit torques (SOTs), the spin Nernst effect can exert thermal SOTs on an adjacent magnetic layer and control the magnetization direction. Here, the thermal SOT caused by the spin Nernst effect is experimentally demonstrated in W/CoFeB/MgO structures. It is found that an in-plane temperature gradient across the sample generates a magnetic torque and modulates the switching field of the perpendicularly magnetized CoFeB. The W thickness dependence suggests that the torque originates mainly from thermal spin currents induced in W. Moreover, the thermal SOT reduces the critical current for SOT-induced magnetization switching, demonstrating that it can be utilized to control the magnetization in spintronic devices.

8.
Anal Chem ; 92(9): 6462-6469, 2020 05 05.
Article in English | MEDLINE | ID: mdl-32267142

ABSTRACT

The field of terminal proteomics is limited in that it is optimized for large-scale analysis via multistep processes involving liquid chromatography. Here, we present an integrated N-terminal peptide enrichment method (iNrich) that can handle as little as 25 µg of cell lysate via a single-stage encapsulated solid-phase extraction column. iNrich enables simple, rapid, and reproducible sample processing, treatment of a wide range of protein amounts (25 µg ∼ 1 mg), multiplexed parallel sample preparation, and in-stage sample prefractionation using a mixed-anion-exchange filter. We identified ∼5000 N-terminal peptides (Nt-peptides) from only 100 µg of human cell lysate including Nt-formyl peptides. Multiplexed sample preparation facilitated quantitative and robust enrichment of N-terminome with dozens of samples simultaneously. We further developed the method to incorporate isobaric tags such as a tandem mass tag (TMT) and used it to discover novel peptides during ER stress analysis. The iNrich facilitated high-throughput N-terminomics and degradomics at a low cost using commercially available reagents and apparatus, without requiring arduous procedures.


Subject(s)
Peptides/chemistry , Proteome/analysis , Cells, Cultured , Chromatography, Liquid , Humans , Hydrogen-Ion Concentration , Solid Phase Extraction , Tandem Mass Spectrometry
9.
Front Plant Sci ; 11: 64, 2020.
Article in English | MEDLINE | ID: mdl-32117399

ABSTRACT

Plants have two endosymbiotic organelles, chloroplast and mitochondrion. Although they have their own genomes, proteome assembly in these organelles depends on the import of proteins encoded by the nuclear genome. Previously, we elucidated the general design principles of chloroplast and mitochondrial targeting signals, transit peptide, and presequence, respectively, which are highly diverse in primary structure. Both targeting signals are composed of N-terminal specificity domain and C-terminal translocation domain. Especially, the N-terminal specificity domain of mitochondrial presequences contains multiple arginine residues and hydrophobic sequence motif. In this study we investigated whether the design principles of plant mitochondrial presequences can be applied to those in other eukaryotic species. We provide evidence that both presequences and import mechanisms are remarkably conserved throughout the species. In addition, we present evidence that the N-terminal specificity domain of presequence might have evolved from the bacterial TAT (twin-arginine translocation) signal sequence.

10.
BMB Rep ; 52(3): 163-164, 2019 Mar.
Article in English | MEDLINE | ID: mdl-30885288

ABSTRACT

The ribosomal synthesis of proteins in the eukaryotic cytosol has always been thought to start from the unformylated N-terminal (Nt) methionine (Met). In contrast, in virtually all nascent proteins in bacteria and eukaryotic organelles, such as mitochondria and chloroplasts, Nt-formyl-methionine (fMet) is the first building block of ribosomal synthesis. Through extensive approaches, including mass spectrometric analyses of the N-termini of proteins and molecular genetic techniques with an affinity-purified antibody for Nt-formylation, we investigated whether Nt-formylated proteins could also be produced and have their own metabolic fate in the cytosol of a eukaryote, such as yeast Saccharomyces cerevisiae. We discovered that Nt-formylated proteins could be generated in the cytosol by yeast mitochondrial formyltransferase (Fmt1). These Nt-formylated proteins were massively upregulated in the stationary phase or upon starvation for specific amino acids and were crucial for the adaptation to specific stresses. The stress-activated kinase Gcn2 was strictly required for the upregulation of Nt-formylated proteins by regulating the activity of Fmt1 and its retention in the cytosol. We also found that the Nt-fMet residues of Nt-formylated proteins could be distinct N-terminal degradation signals, termed fMet/N-degrons, and that Psh1 E3 ubiquitin ligase mediated the selective destruction of Nt-formylated proteins as the recognition component of a novel eukaryotic fMet/N-end rule pathway, termed fMet/N-recognin. [BMB Reports 2019; 52(3): 163-164].


Subject(s)
Hydroxymethyl and Formyl Transferases/physiology , Protein Biosynthesis/physiology , Amino Acids/metabolism , Cytosol/metabolism , Eukaryotic Cells/metabolism , Hydroxymethyl and Formyl Transferases/metabolism , Methionine/metabolism , Mitochondria/metabolism , Proteins/metabolism , Proteolysis , Saccharomyces cerevisiae/metabolism , Saccharomyces cerevisiae Proteins/metabolism , Ubiquitin-Protein Ligases/metabolism , Up-Regulation
11.
J Biol Chem ; 294(12): 4464-4476, 2019 03 22.
Article in English | MEDLINE | ID: mdl-30674553

ABSTRACT

All organisms begin protein synthesis with methionine (Met). The resulting initiator Met of nascent proteins is irreversibly processed by Met aminopeptidases (MetAPs). N-terminal (Nt) Met excision (NME) is an evolutionarily conserved and essential process operating on up to two-thirds of proteins. However, the universal function of NME remains largely unknown. MetAPs have a well-known processing preference for Nt-Met with Ala, Ser, Gly, Thr, Cys, Pro, or Val at position 2, but using CHX-chase assays to assess protein degradation in yeast cells, as well as protein-binding and RT-qPCR assays, we demonstrate here that NME also occurs on nascent proteins bearing Met-Asn or Met-Gln at their N termini. We found that the NME at these termini exposes the tertiary destabilizing Nt residues (Asn or Gln) of the Arg/N-end rule pathway, which degrades proteins according to the composition of their Nt residues. We also identified a yeast DNA repair protein, MQ-Rad16, bearing a Met-Gln N terminus, as well as a human tropomyosin-receptor kinase-fused gene (TFG) protein, MN-TFG, bearing a Met-Asn N terminus as physiological, MetAP-processed Arg/N-end rule substrates. Furthermore, we show that the loss of the components of the Arg/N-end rule pathway substantially suppresses the growth defects of naa20Δ yeast cells lacking the catalytic subunit of NatB Nt acetylase at 37 °C. Collectively, the results of our study reveal that NME is a key upstream step for the creation of the Arg/N-end rule substrates bearing tertiary destabilizing residues in vivo.


Subject(s)
Arginine/metabolism , Methionine/metabolism , Protein Biosynthesis , Saccharomyces cerevisiae/metabolism , Humans , Proteins/chemistry , Proteins/metabolism , Proteolysis
12.
Science ; 362(6418)2018 11 30.
Article in English | MEDLINE | ID: mdl-30409808

ABSTRACT

In bacteria, nascent proteins bear the pretranslationally generated N-terminal (Nt) formyl-methionine (fMet) residue. Nt-fMet of bacterial proteins is a degradation signal, termed fMet/N-degron. By contrast, proteins synthesized by cytosolic ribosomes of eukaryotes were presumed to bear unformylated Nt-Met. Here we found that the yeast formyltransferase Fmt1, although imported into mitochondria, could also produce Nt-formylated proteins in the cytosol. Nt-formylated proteins were strongly up-regulated in stationary phase or upon starvation for specific amino acids. This up-regulation strictly required the Gcn2 kinase, which phosphorylates Fmt1 and mediates its retention in the cytosol. We also found that the Nt-fMet residues of Nt-formylated proteins act as fMet/N-degrons and identified the Psh1 ubiquitin ligase as the recognition component of the eukaryotic fMet/N-end rule pathway, which destroys Nt-formylated proteins.


Subject(s)
Amino Acids/deficiency , Hydroxymethyl and Formyl Transferases/metabolism , N-Formylmethionine/metabolism , Proteolysis , Saccharomyces cerevisiae Proteins/metabolism , Saccharomyces cerevisiae/metabolism , Azides/pharmacology , Cold Temperature , Cytosol/metabolism , Metabolic Networks and Pathways , Mitochondria/enzymology , N-Formylmethionine/chemistry , Peptide Elongation Factors/metabolism , Phosphorylation , Protein Serine-Threonine Kinases/metabolism , Saccharomyces cerevisiae/drug effects , Saccharomyces cerevisiae/enzymology , Ubiquitin-Protein Ligases/metabolism , Up-Regulation
13.
Sci Rep ; 8(1): 11337, 2018 Jul 27.
Article in English | MEDLINE | ID: mdl-30054593

ABSTRACT

We investigated the temperature distribution induced by laser irradiation of ultrathin magnetic films by applying a finite element method (FEM) to the finite difference time domain (FDTD) representation for the analysis of thermal induced spin currents. The dependency of the thermal gradient (∇T) of ultrathin magnetic films on material parameters, including the reflectivity and absorption coefficient were evaluated by examining optical effects, which indicates that reflectance (R) and the apparent absorption coefficient (α*) play important roles in the calculation of ∇T for ultrathin layers. The experimental and calculated values of R and α* for the ultrathin magnetic layers irradiated by laser-driven heat sources estimated using the combined FDTD and FEM method are in good agreement for the amorphous CoFeB and crystalline Co layers of thicknesses ranging from 3~20 nm. Our results demonstrate that the optical parameters are crucial for the estimation of the temperature gradient induced by laser illumination for the study of thermally generated spin currents and related phenomena.

14.
World Neurosurg ; 104: 1049.e11-1049.e15, 2017 Aug.
Article in English | MEDLINE | ID: mdl-28552697

ABSTRACT

BACKGROUND: A 52-year-old woman had a 20-month history of progressive radiating pain in the left arm and numbness on C7 dermatome. CASE DESCRIPTION: On physical examination, left head rotation aggravated the radiculopathic pain. For an anatomic diagnosis of the vertebral artery and nerve root, magnetic resonance angiography was performed (computed tomography angiography was not possible because of her dye allergy history). Magnetic resonance angiography showed a left vertebral artery loop entering at the C6-7 intervertebral foramen. Surgical microvascular decompression was performed by an anterior cervical approach and the loop of the artery was fixed using a sling technique. Postoperative computed tomography angiography showed that the left vertebral artery was retracted anteriorly and the C7 nerve root was decompressed in the intervertebral foramen at the left C6-7 level. CONCLUSIONS: The patient's radiculopathic symptoms were improved and especially the aggravated pain by left head rotation subsided dramatically.


Subject(s)
Cervical Vertebrae , Microvascular Decompression Surgery/methods , Radiculopathy/surgery , Vertebral Artery/surgery , Cerebral Angiography , Computed Tomography Angiography , Female , Humans , Magnetic Resonance Angiography , Magnetic Resonance Imaging , Middle Aged , Radiculopathy/diagnostic imaging , Vertebral Artery/diagnostic imaging
15.
Mol Cells ; 39(3): 169-78, 2016 Mar.
Article in English | MEDLINE | ID: mdl-26883906

ABSTRACT

Although Nα-terminal acetylation (Nt-acetylation) is a pervasive protein modification in eukaryotes, its general functions in a majority of proteins are poorly understood. In 2010, it was discovered that Nt-acetylation creates a specific protein degradation signal that is targeted by a new class of the N-end rule proteolytic system, called the Ac/N-end rule pathway. Here, we review recent advances in our understanding of the mechanism and biological functions of the Ac/N-end rule pathway, and its crosstalk with the Arg/N-end rule pathway (the classical N-end rule pathway).


Subject(s)
Protein Processing, Post-Translational , Proteins/metabolism , Acetylation , Humans , Proteins/chemistry , Proteolysis , Signal Transduction
16.
Science ; 347(6227): 1249-1252, 2015 Mar 13.
Article in English | MEDLINE | ID: mdl-25766235

ABSTRACT

Rgs2, a regulator of G proteins, lowers blood pressure by decreasing signaling through Gαq. Human patients expressing Met-Leu-Rgs2 (ML-Rgs2) or Met-Arg-Rgs2 (MR-Rgs2) are hypertensive relative to people expressing wild-type Met-Gln-Rgs2 (MQ-Rgs2). We found that wild-type MQ-Rgs2 and its mutant, MR-Rgs2, were destroyed by the Ac/N-end rule pathway, which recognizes N(α)-terminally acetylated (Nt-acetylated) proteins. The shortest-lived mutant, ML-Rgs2, was targeted by both the Ac/N-end rule and Arg/N-end rule pathways. The latter pathway recognizes unacetylated N-terminal residues. Thus, the Nt-acetylated Ac-MX-Rgs2 (X = Arg, Gln, Leu) proteins are specific substrates of the mammalian Ac/N-end rule pathway. Furthermore, the Ac/N-degron of Ac-MQ-Rgs2 was conditional, and Teb4, an endoplasmic reticulum (ER) membrane-embedded ubiquitin ligase, was able to regulate G protein signaling by targeting Ac-MX-Rgs2 proteins for degradation through their N(α)-terminal acetyl group.


Subject(s)
RGS Proteins/metabolism , Acetylation , Amino Acid Sequence , GTP-Binding Protein alpha Subunits, Gq-G11/metabolism , HEK293 Cells , HeLa Cells , Humans , Membrane Proteins/genetics , Membrane Proteins/metabolism , Mutant Proteins/chemistry , Mutant Proteins/metabolism , Protein Processing, Post-Translational , Protein Stability , Proteolysis , RGS Proteins/chemistry , RGS Proteins/genetics , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Signal Transduction , Ubiquitin-Protein Ligases/genetics , Ubiquitin-Protein Ligases/metabolism , Ubiquitination
17.
J Microbiol ; 52(10): 842-8, 2014 Oct.
Article in English | MEDLINE | ID: mdl-25163837

ABSTRACT

The Saccharomyces cerevisiae NatB N-terminal acetylase contains a catalytic subunit Naa20 and an auxiliary subunit Naa25. To elucidate the cellular functions of the NatB, we utilized the Synthetic Genetic Array to screen for genes that are essential for cell growth in the absence of NAA20. The genome-wide synthetic lethal screen of NAA20 identified genes encoding for serine/threonine protein kinase Vps15, 1,3-beta-glucanosyltransferase Gas5, and a catabolic repression regulator Mig3. The present study suggests that the catalytic activity of the NatB N-terminal aceytase is involved in vacuolar protein sorting and cell wall maintenance.


Subject(s)
Gene Deletion , Genes, Essential , N-Terminal Acetyltransferase B/genetics , N-Terminal Acetyltransferase B/metabolism , Saccharomyces cerevisiae Proteins/genetics , Saccharomyces cerevisiae Proteins/metabolism , Saccharomyces cerevisiae/enzymology , Saccharomyces cerevisiae/physiology , Catalytic Domain/genetics , Genes, Fungal , Glucosyltransferases/genetics , Glucosyltransferases/metabolism , Repressor Proteins/genetics , Repressor Proteins/metabolism , Saccharomyces cerevisiae/growth & development , Vacuolar Sorting Protein VPS15/genetics , Vacuolar Sorting Protein VPS15/metabolism
19.
Proteins ; 82(6): 1093-8, 2014 Jun.
Article in English | MEDLINE | ID: mdl-24356916

ABSTRACT

BldD regulates transcription of key developmental genes in Streptomyces coelicolor. While the N-terminal domain is responsible for both dimerization and DNA binding, the structural and functional roles of the C-terminal domain (CTD) remain largely unexplored. Here, the solution structure of the BldD-CTD shows a novel winged-helix domain fold not compatible with DNA binding, due to the negatively charged surface and presence of an additional helix. Meanwhile, a small elongated groove with conserved hydrophobic patches surrounded by charged residues suggests that the BldD-CTD could be involved in protein-protein interactions that provide transcriptional regulation.


Subject(s)
Bacterial Proteins/chemistry , Streptomyces coelicolor , Winged-Helix Transcription Factors/chemistry , Amino Acid Sequence , Conserved Sequence , Hydrophobic and Hydrophilic Interactions , Models, Molecular , Molecular Sequence Data , Nuclear Magnetic Resonance, Biomolecular , Protein Interaction Domains and Motifs , Protein Structure, Secondary
20.
J Food Sci ; 78(3): C425-31, 2013 Mar.
Article in English | MEDLINE | ID: mdl-23425135

ABSTRACT

UNLABELLED: Enzymatic browning remains a problem for the fruit and vegetable industry, especially new emerging markets like pre-cuts. A crude inhibitor from blue mussel (Mytilus edulis) showed broad inhibition for apple (58%), mushroom (32%), and potato (44%) polyphenol oxidase (PPO) and was further characterized. Inhibition increased as the concentration of inhibitor increased in the reaction mixture eventually leveling off at a maximum inhibition of 92% for apple PPO. The inhibitor was capable of bleaching the brown color formed in the reaction mixture with apple PPO. Identification of the inhibitor by mass spectrometry and high-performance liquid chromatography revealed it to be hypotaurine (C2 H7 NO2 S). Hypotaurine and other sulfinic acid analogs (methane and benzene sulfinic acids) showed very good inhibition for apple PPO at various concentrations with the highest inhibition occurring at 500 µM for hypotaurine (89%), methane sulfinic acid (100%), and benzene sulfinic acid (100%). PRACTICAL APPLICATION: An inhibitor found in the expressed liquid from blue mussel shows very good inhibition on enzymatic browning. Since this enzyme is responsible for losses to the fruit and vegetable industry, natural inhibitors that prevent browning would be valuable. Finding alternative chemistries that inhibit browning and understanding their mode of action would be beneficial to the fruit and vegetable industries and their segments such as pre-cuts, juices, and so on. Inhibitors from products ingested by consumers are more acceptable as natural ingredients.


Subject(s)
Catechol Oxidase/antagonists & inhibitors , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/isolation & purification , Mytilus edulis/chemistry , Agaricales/enzymology , Animals , Chromatography, High Pressure Liquid , Fruit/enzymology , Malus/enzymology , Solanum tuberosum/enzymology , Vegetables/enzymology
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