ABSTRACT Thermomonospora fusca E4 is an unusual 90.4-kDa endocellulase comprised of a catalytic domain (CD), internal family IIIc cellulose binding (CBD), fibronectinlike domain, and II CBD. Constructs containing the CD alone (E4-51), plus CBD (E4-68), (E4-74) were made by using recombinant DNA techniques. The activities each purified protein on bacterial microcrystalline (BMCC), filter paper, swollen cellulose, carboxymethyl measured. Only whole enzyme, E4-90, could reach target digestion...

ABSTRACT In C 3 plants, photorespiration is an energy expensive pathway that competes with photosynthetic CO 2 assimilation and releases into the atmosphere, potentially reducing plant productivity by 20%–50%. Consequently, flux through has been recognized as a major way to improve crop carbon fixation productivity. While current research efforts in engineering are mainly based on modification of chloroplast glycolate metabolic steps, only limited studies have explored optimizations other...

Electrospray ionization mass spectrometry (ESI-MS) has proven to be an extremely powerful tool for studying biomolecular structures and noncovalent interactions. Here we report a method using fully automated, chip-based nanoESI-MS system determine the dissociation constants (Kd) complexes of two different proteins with their ligands. The automated nanoelectrospray system, consisting NanoMate ESI chip, serves functionally as combination autosampler source. This provides all advantages...

Cysteine dioxygenase (CDO) catalyzes the conversion of cysteine to cysteinesulfinic acid and is important in regulation intracellular levels mammals provision oxidized metabolites such as sulfate taurine. Several crystal structure studies mammalian CDO have shown that there a cross-linked cofactor present active site enzyme. The consists thioether bond between gamma-sulfur residue 93 aromatic side chain tyrosine 157. exact requirements for synthesis contribution catalytic activity enzyme yet...

Cold-induced sweetening in potato tubers is a costly problem for the food industry. To systematically identify proteins associated with this process, we employed comparative proteomics approach using isobaric, stable isotope coded labels to compare proteomes of after 0 and 5 months storage at °C. We evaluated both high pH reverse phase (hpRP) liquid chromatography (LC) off-gel electrophoresis (OGE) as first dimension fractionation methods followed by nanoLC–MS/MS, two performance mass...

Despite the relationship between melatonin and aging, overall changes regulation of proteome profiling by long-term exposure during leaf senescence is not well understood. In this study, in Malus hupehensis plants was delayed when exogenous regularly applied to roots for 2 months compared with natural senescence. Proteins samples 0 50 day both treatments were extracted labeled TMT regents before being examined via NanoLC-MS/MS. The proteomics data showed that 622 309 proteins altered...

Over the past decades, substantial advances have been made in both early diagnosis and accurate prognosis of numerous cancers because impressive development novel proteomic strategies. Selenium (Se) is an essential trace element humans animals. Se deficiency could lead to Keshan disease humans, mulberry heart pigs damage tissues including cardiac injury, apoptosis liver, reduction immune responses spleen cerebral lesions chickens. However, it well know that plasma biomarkers are not specific...

Nonlinear kinetics are commonly observed in the enzymatic hydrolysis of cellulose. This nonlinearity could be explained by any or all following three factors: enzyme inactivation, product inhibition, substrate heterogeneity. In this study, four different approaches were applied to test above hypotheses using two Thermomonospora fusca endocellulases, E2 and E5. The lack stimulation cellulase activity β-glucosidase rules out possibility inhibition as a cause nonlinearity. results from other...

Protein tyrosine phosphatases (PTPs) regulate a broad range of cellular processes including proliferation, differentiation, migration, apoptosis, and immune responses. Dysfunction PTP activity is associated with cancers, metabolic syndromes, autoimmune disorders. Consequently, small molecule inhibitors should serve not only as powerful tools to delineate the physiological roles these enzymes in vivo but also lead compounds for therapeutic development. We describe novel stepwise...

Glycolate oxidase (GLO) is a key enzyme for photorespiration in plants. Previous studies have demonstrated that suppression of GLO causes photosynthetic inhibition, and the accumulated glycolate with deactivated Rubisco likely involved regulation. Using isolated chloroplasts, it has been found only glyoxylate can effectively inactivate meanwhile inhibit photosynthesis, but little vivo evidence acquired reported. In this study, we generated transgenic rice (Oryza sativa) plants being...

Photorespiration begins with the oxygenation reaction catalyzed by 1,5-bisphosphate carboxylase/oxygenase (Rubisco) and serves as a repair pathway for carbon retrieval converting 2-phosphoglycolate to 3-phosphoglycerate, allowing plants thrive in an oxygen-rich environment. metabolism is intimately linked plant primary metabolism, particularly nitrogen assimilation, cellular redox equilibrium, such interactions are dynamically regulated environmental changes. Although basic genetics...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTCharacterization of a Thermomonospora fusca exocellulaseSheng Zhang, Guifang Lao, and David B. WilsonCite this: Biochemistry 1995, 34, 10, 3386–3395Publication Date (Print):March 1, 1995Publication History Published online1 May 2002Published inissue 1 March 1995https://pubs.acs.org/doi/10.1021/bi00010a030https://doi.org/10.1021/bi00010a030research-articleACS PublicationsRequest reuse permissionsArticle Views529Altmetric-Citations69LEARN ABOUT THESE...

The bifunctional P-protein, which plays a central role in <i>Escherichia coli</i> phenylalanine biosynthesis, contains two catalytic domains (chorismate mutase and prephenate dehydratase activities) as well one R-domain (for feedback inhibition by phenylalanine). Six genes coding for P-protein or subdomains were constructed successfully expressed. Proteins containing residues 1–285 1–300 retained full activity, but exhibited no inhibition. 101–386 101–300 lacked activity. Fluorescence...

Protein tyrosine phosphatases (PTPs) constitute a large family of signaling enzymes that control the cellular levels protein phosphorylation. A detailed understanding PTP functions in normal physiology and pathogenic conditions has been hampered by absence PTP-specific, cell-permeable small-molecule agents. We present stepwise focused library approach transforms weak general non-hydrolyzable pTyr mimetic (F2Pmp, phosphonodifluoromethyl phenylalanine) into highly potent selective inhibitor...

Abstract Background The insect cell line is a critical component in the production of recombinant proteins baculovirus expression system and new lines hold promise increasing both quantity quality protein production. Results Seventy were established by single-cell cloning from primary culture cells derived eggs black witch moth ( Ascalapha odorata ; Lepidoptera, Noctuidae). Among 8 rapidly growing lines, 38 (Ao38) was selected for further analysis, based on susceptibility to AcMNPV infection...

// Chen Jue 1, 2, 3, * , Cui Lin Zhang Zhisheng Qian Yayun 1 Jin Feng Zhao Min Wang Haibo Shi Youyang Tadashi Hisamitsu 3 Ishikawa Shintaro Guo Shiyu Liu Yanqing Institution of Combining Chinese Traditional and Western Medicine, Medical College, Yangzhou University, Yangzhou, Jiangsu, China 2 Department Oncology, the Second Peoples Hospital Taizhou affiliated to Taizhou, Physiology, School Showa Tokyo, Japan These authors have contributed equally this work Correspondence to: Yanqing, email:...

NCA1 (NO CATALASE ACTIVITY 1) was recently identified in Arabidopsis as a chaperone protein to regulate catalase (CAT) activity through maintaining the folding of CAT. The gene exists mainly higher plants; some plants, such Arabidopsis, contain only one gene, whereas others rice harbor two copies. It is not yet understood whether and how both isoforms have functioned CAT those two-copy-containing plant species. In this study, we first noticed that spatiotemporal expression patterns NCA1a...

Polyploidy plays an important role in crop improvement. Polyploid plants, particularly those produced through unreduced gametes (2n gametes), show increased organ size, improved buffering capacity for deleterious mutations, and enhanced heterozygosity heterosis. Induced polyploidy has been widely used improving floriculture crops, however, there are few reported sexual polyploid plants the industry. This study evaluated nine cultivars of Cymbidium Swartz discovered that 2n male occurred this...

In this study, a four-enzyme cascade pathway was developed and reconstructed in vivo for the production of D-p-hydroxyphenylglycine (D-HPG), valuable intermediate used to produce β-lactam antibiotics fine-chemical synthesis, from L-tyrosine. pathway, catalytic conversion 4-hydroxyphenylglyoxalate by meso-diaminopimelate dehydrogenase Corynebacterium glutamicum (CgDAPDH) identified as rate-limiting step, followed application mechanism-guided "conformation rotation" strategy decrease...

Abstract Background Glycolate oxidase (GLO) is not only a key enzyme in photorespiration but also major engine for H 2 O production plants. Catalase (CAT)-dependent decomposition has been previously reported to be involved the regulation of IAA biosynthesis. However, it still known which mechanism contributed regulation. Results In this study, we found that glo mutants rice, as levels decreased contents significantly increased, whereas high CO abolished difference and between WT. Further...