With the development of global economy, the dramatically increased production of polyethylene terephthalate (PET) plastics has led to a remarkably increased amount of plastic waste. PET waste can be treated by landfill, incineration, or biodegradation. While landfilling and incineration may cause secondary pollution, biodegradation has since received increased attentions due to its environmental friendliness. Recent studies have indicated that the carbohydrate binding module (CBM) can effectively enhance the binding of PET degrading enzymes to PET, and consequently increasing PET degradation rate. Here we constructed a fusion protein BaCBM2-Tfuc containing the BaCBM2 from Bacillus anthraci and the cutinase Tfuc from Thermobifida fusca, by megaprimer PCR of whole plasmids (MEGAWHOP). Notabaly, the PET film degradation efficiency (at 60 ℃) of BaCBM2-Tfuc was 2.8 times that of Tfuc. This study may provide technical support for constructing fusion proteins capable of efficiently degrading PET.
Carbohydrates ; Carboxylic Ester Hydrolases ; Polyethylene Terephthalates ; Thermobifida

With the decrease of forest timber resources, the recycling of waste paper has received increasing attention. However, the stickies produced in the process of waste paper recycling may negatively affect the production of recycled paper. The biological decomposition of stickies, which has the advantages of high efficiency, high specificity and pollution-free, is achieved mainly through the enzymatic cleavage of the ester bond in the stickies components to prevent flocculation. Cutinase is a serine esterase that can degrade some components of the stickies. Previous research indicated that the anchor peptide tachystatin A2 (TA2) is able to bind polyurethane. In this study, the cutinase HiC derived from Humicola insolens was used to construct a fusion protein HiC-TA2 by megaprimer PCR of the whole plasmid (MEGAWHOP). The enzymatic properties and the degradation efficiency of the fusion protein on poly(ethyl acrylate) (PEA), a model substrate of stickies component, were determined. The results showed that the degradation efficiency, the size decrease of PEA particle, and the amount of ethanol produced by HiC-TA2 were 1.5 times, 6.8 times, and 1.4 times of that by HiC, respectively. These results demonstrated that TA2 improved the degradation efficiency of HiC on PEA. This study provides a useful reference for biological decomposition of stickies produced in the process of recycled paper production.
Carboxylic Ester Hydrolases/genetics* ; Fungal Genus Humicola ; Polyurethanes

Cutinase can degrade aliphatic and aromatic polyesters, as well as polyethylene terephthalate. Lack of commercially available cutinase calls for development of cost-effective production of efficient cutinase. In this study, eight cutinase genes were cloned from Sclerotinia sclerotiorum. The most active gene SsCut-52 was obtained by PCR combined with RT-PCR, expressed in Escherichia coli BL21 and purified by Ni-NTA affinity chromatography to study its characteristics and pathogenicity. Sscut-52 had a total length of 768 bp and 17 signal peptides at the N terminals. Phylogenetic analysis showed that its amino acid sequence had the highest homology with Botrytis keratinase cutinase and was closely related to Rutstroemia cutinase. Sscut-52 was highly expressed during the process of infecting plants by Sclerotinia sclerotiorum. Moreover, the expression level of Sscut-52 was higher than those of other cutinase genes in the process of sclerotia formation from mycelium. The heterologously expressed cutinase existed in the form of inclusion body. The renatured SsCut-52 was active at pH 4.0-10.0, and mostly active at pH 6.0, with a specific activity of 3.45 U/mg achieved. The optimum temperature of SsCut-52 was 20-30 ℃, and less than 60% of the activity could be retained at temperatures higher than 50 ℃. Plant leaf infection showed that SsCut-52 may promote the infection of Banlangen leaves by Sclerotinia sclerotiorum.
Ascomycota/genetics* ; Carboxylic Ester Hydrolases ; Cloning, Molecular ; Phylogeny

Alzheimer's disease is a chronic neurodegenerative disorder with no curative treatment. The commercially available drugs, which target acetylcholinesterase, are not satisfactory. The aim of this study was to investigate the cholinesterase inhibitory activity of Solenostemma argel aerial part. Eight compounds were isolated and identified by NMR: kaempferol-3-O-glucopyranoside (1), kaempferol (2), kaempferol-3-glucopyranosyl(1→6)rhamnopyranose (3) p-hydroxybenzoic acid (4), dehydrovomifoliol (5), 14,15-dihydroxypregn-4-ene-3,20-dione (6), 14,15-dihydroxy-pregn-4-ene-3,20-dione-15β-D-glucopyranoside (7) and solargin I (8). Two of them (compounds 2 and 3) could inhibit over 50 % of butyrylcholinesterase activity at 100 µM. Compound (2) displayed the highest inhibitory effect against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) with a slight selectivity towards the latter. Molecular docking studies supported the in vitro results and revealed that (2) had made several hydrogen and π-π stacking interactions which could explain the compound potency to inhibit AChE and BChE.
Acetylcholinesterase ; Alzheimer Disease ; Butyrylcholinesterase ; Cholinesterases ; Hydrogen ; In Vitro Techniques ; Neurodegenerative Diseases

BACKGROUND: Recurrent aphthous stomatitis (RAS) is a chronic relapsing inflammatory disorder of the oral mucosa with unknown etiology. Oxidative stress (OS) is suggested to play a main role in the etiopathogenesis in RAS. OBJECTIVE: In this study, we hypothesize that a systemic OS is present in patients with RAS. METHODS: Forty-four patients with active RAS lesions and 38 healthy controls were being included in the study. Serum total oxidant status (TOS), total antioxidant status (TAS), oxidative stress index (OSI), and paraoxonase 1 arylesterase (ARES) activity were being determined. RESULTS: RAS patients had significantly lower TAS levels and higher TOS and OSI values than controls. The patients had a lower ARES activity when compared to healthy controls. No correlations were observed between OS parameters and age, gender, duration of disease or frequency of RAS attacks per month. CONCLUSION: A systemic OS is determined with an imbalance in oxidant/antioxidant status and lower ARES activity in RAS. Systemic OS may have an important role in the pathogenesis of RAS formation.
Aryldialkylphosphatase ; Carboxylic Ester Hydrolases ; Humans ; Mouth Mucosa ; Oxidative Stress ; Stomatitis, Aphthous

BACKGROUND: Recurrent aphthous stomatitis (RAS) is a chronic relapsing inflammatory disorder of the oral mucosa with unknown etiology. Oxidative stress (OS) is suggested to play a main role in the etiopathogenesis in RAS. OBJECTIVE: In this study, we hypothesize that a systemic OS is present in patients with RAS. METHODS: Forty-four patients with active RAS lesions and 38 healthy controls were being included in the study. Serum total oxidant status (TOS), total antioxidant status (TAS), oxidative stress index (OSI), and paraoxonase 1 arylesterase (ARES) activity were being determined. RESULTS: RAS patients had significantly lower TAS levels and higher TOS and OSI values than controls. The patients had a lower ARES activity when compared to healthy controls. No correlations were observed between OS parameters and age, gender, duration of disease or frequency of RAS attacks per month. CONCLUSION: A systemic OS is determined with an imbalance in oxidant/antioxidant status and lower ARES activity in RAS. Systemic OS may have an important role in the pathogenesis of RAS formation.
Aryldialkylphosphatase ; Carboxylic Ester Hydrolases ; Humans ; Mouth Mucosa ; Oxidative Stress ; Stomatitis, Aphthous

Ferulic acid esterase (FAE) was used to hydrolyze feruloyl ester linkages between hemicellulose and lignin in natural lignocellulose, and the possibility of FAE accelerating cellulase to hydrolyze steam-exploded rice straw by breaking covalent linkages was studied. When the dosage of FAE was 240 mu/g substrate, the cellulose conversion rate and the weight-loss rate of insoluble substrate at 72 h were respectively 32.00% and 32.77%, more than without using FAE; Cellulose conversion rate and the weight-loss rate of insoluble substrate were respectively 29.85% and 32.48% with FAE (300 mu/g substrate) processing time of 120 min. By comparison of the accessibility and FT-IR spectra of lignocellulosic material treated by different enzyme methods, it indicated that FAE hydrolyzed some ester bonds within it, and improved the accessibility by over 50%. It is concluded that FAE and cellulase have great synergistic effect, and FAE can help cellulase hydrolyze natural lignocellulose and enhance hydrolytic efficiency.
Carboxylic Ester Hydrolases ; metabolism ; Cellulase ; metabolism ; Cellulose ; metabolism ; Oryza ; enzymology ; Plant Stems ; enzymology ; Steam

A new method to express oligomerized feruloyl esterase (FAE) in Pichia pastoris GS115 to improve the catalytic efficiency was developed. It was realized by fusing the foldon domain at the C-terminus of FAE, and the fusion protein was purified by histidine tag. Fusion of the feruloyl esterase with the foldon domain resulted spontaneously forming a trimer FAE to improve the catalytic performance. The oligomerized FAE and monomeric FAE were obtained by purification. The apparent molecular weight of the oligomerized FAE was about 110 kDa, while the monomeric FAE about 40 kDa, and the optimum temperature of the oligomerized FAE was 50 °C, which is the same as the monomeric one. The optimal pH of the oligomerized FAE is 5.0, while the optimal pH of the monomer FAE is 6.0. When compared with the monomeric ones, the catalytic efficiency (kcat/Km) of the oligomerized FAE increased 7.57-folds. The catalytic constant (kcat) of the oligomerized FAE increased 3.42-folds. The oligomerized FAE induced by foldon have advantages in the catalytic performances, which represents a simple and effective enzyme-engineering tool. The method proposed here for improving the catalytic efficiency of FAE would have great potentials for improving the catalytic efficiency of other enzymes.
Carboxylic Ester Hydrolases ; metabolism ; Catalysis ; Molecular Weight ; Pichia ; genetics ; metabolism ; Polymerization ; Protein Engineering ; Substrate Specificity

Pectin methylesterase (PME) is an important pectinase that hydrolyzes methyl esters in pectin to release methanol and reduce the degree of methylation of pectin. At present, it has broad application prospects in food processing, tea beverage, paper making and other production processes. With the in-depth study of PME, the crystal structures with different sources have been reported. Analysis of these resolved crystal structures reveals that PME belongs to the right-hand parallel β-helix structure, and its catalytic residues are two aspartic acids and a glutamine, which play the role of general acid-base, nucleophile and stable intermediate, in the catalytic process. At the same time, the substrate specificity is analyzed to understand the recognition mechanism of the substrate and active sites. This paper systematically reviews these related aspects.
Carboxylic Ester Hydrolases ; chemistry ; metabolism ; Catalytic Domain ; Crystallography ; Pectins ; metabolism ; Protein Structure, Tertiary ; Substrate Specificity

OBJECTIVE: To explore the clinical phenotype and genetic features of two children with MEGDEL syndrome due to variants of the SERAC1 gene. METHODS: Two children who had presented at the Fujian Medical University Union Hospital respectively on July 14, 2020 and July 28, 2018 were selected as the study subjects. Clinical features and results of genetic testing were retrospectively analyzed. RESULTS: Both children had featured developmental delay, dystonia and sensorineural deafness, along with increased urine 3-methylglutaric acid levels. Magnetic resonance imaging revealed changes similar to Leigh-like syndrome. Gene sequencing revealed that both children have harbored pathogenic compound heterozygous variants of the SERAC1 gene, including c.1159C>T and c.442C>T in child 1, and c.1168C>T and exons 4~9 deletion in child 2. CONCLUSION Children with MEGDEL syndrome due to SERAC1 gene variants have variable clinical genotypes. Delineation of its clinical characteristics and typical imaging changes can facilitate early diagnosis and treatment. Discovery of the novel variants has also enriched the spectrum of SERAC1 gene variants.
Humans ; Retrospective Studies ; Metabolism, Inborn Errors ; Hearing Loss, Sensorineural/genetics* ; Dystonia ; Carboxylic Ester Hydrolases