Biochemical properties of extracellular protease from Staphylococcus epidermidis isolated from Harbin dry sausages and its hydrolysis of meat protein

2021 ◽  
pp. 101130
Author(s):  
Hui Wang ◽  
Jinming Liu ◽  
Qian Chen ◽  
Baohua Kong ◽  
Fangda Sun
Keyword(s):  
Staphylococcus Epidermidis ◽  
Biochemical Properties ◽  
Extracellular Protease ◽  
Meat Protein ◽  
Dry Sausages ◽  
Hydrolysis Of

scholarly journals Mutating His29, His125, His133 or His158 abolishes glycosylphosphatidylinositol-specific phospholipase D catalytic activity

2005 ◽  
Vol 391 (2) ◽  
pp. 285-289 ◽  
Author(s):  
Nandita S. Raikwar ◽  
Rosario F. Bowen ◽  
Mark A. Deeg
Keyword(s):  
Catalytic Activity ◽  
Phospholipase D ◽  
Catalytic Domain ◽  
Computer Modelling ◽  
Critical Role ◽  
Biochemical Properties ◽  
Catalytic Site ◽  
Wild Type ◽  
Histidine Residues ◽  
Hydrolysis Of

Glycosylphosphatidylinositol (GPI)-specific phospholipase D (GPI-PLD) specifically cleaves GPIs. This phospholipase D is a secreted protein consisting of two domains: an N-terminal catalytic domain and a predicted C-terminal β-propeller. Although the biochemical properties of GPI-PLD have been extensively studied, its catalytic site has not been identified. We hypothesized that a histidine residue(s) may play a critical role in the catalytic activity of GPI-PLD, based on the observations that (i) Zn2+, which utilizes histidine residues for binding, is required for GPI-PLD catalytic activity, (ii) a phosphohistidine intermediate is involved in phospholipase D hydrolysis of phosphatidylcholine, (iii) computer modelling suggests a catalytic site containing histidine residues, and (iv) our observation that diethyl pyrocarbonate, which modifies histidine residues, inhibits GPI-PLD catalytic activity. Individual mutation of the ten histidine residues to asparagine in the catalytic domain of murine GPI-PLD resulted in three general phenotypes: not secreted or retained (His56 or His88), secreted with catalytic activity (His34, His81, His98 or His219) and secreted without catalytic activity (His29, His125, His133 or His158). Changing His133 but not His29, His125 or His158 to Cys resulted in a mutant that retained catalytic activity, suggesting that at least His133 is involved in Zn2+ binding. His133 and His158 also retained the biochemical properties of wild-type GPI-PLD including trypsin cleavage pattern and phosphorylation by protein kinase A. Hence, His29, His125, His133 and His158 are required for GPI-PLD catalytic activity.

scholarly journals Comparative Analyses of Two Thermophilic Enzymes Exhibiting both β-1,4 Mannosidic and β-1,4 Glucosidic Cleavage Activities from Caldanaerobius polysaccharolyticus

2010 ◽  
Vol 192 (16) ◽  
pp. 4111-4121 ◽  
Author(s):  
Yejun Han ◽  
Dylan Dodd ◽  
Charles W. Hespen ◽  
Samuel Ohene-Adjei ◽  
Charles M. Schroeder ◽  
...  
Keyword(s):  
Catalytic Domain ◽  
Biochemical Properties ◽  
Degree Of Polymerization ◽  
Glycoside Hydrolase Family ◽  
Carbohydrate Binding ◽  
Biofuel Industry ◽  
Carbohydrate Binding Modules ◽  
Mannanase Activity ◽  
Hydrolysis Of ◽  
Derivatives Of

ABSTRACT The hydrolysis of polysaccharides containing mannan requires endo-1,4-β-mannanase and 1,4-β-mannosidase activities. In the current report, the biochemical properties of two endo-β-1,4-mannanases (Man5A and Man5B) from Caldanaerobius polysaccharolyticus were studied. Man5A is composed of an N-terminal signal peptide (SP), a catalytic domain, two carbohydrate-binding modules (CBMs), and three surface layer homology (SLH) repeats, whereas Man5B lacks the SP, CBMs, and SLH repeats. To gain insights into how the two glycoside hydrolase family 5 (GH5) enzymes may aid the bacterium in energy acquisition and also the potential application of the two enzymes in the biofuel industry, two derivatives of Man5A (Man5A-TM1 [TM1 stands for truncational mutant 1], which lacks the SP and SLH repeats, and Man5A-TM2, which lacks the SP, CBMs, and SLH repeats) and the wild-type Man5B were biochemically analyzed. The Man5A derivatives displayed endo-1,4-β-mannanase and endo-1,4-β-glucanase activities and hydrolyzed oligosaccharides with a degree of polymerization (DP) of 4 or higher. Man5B exhibited endo-1,4-β-mannanase activity and little endo-1,4-β-glucanase activity; however, this enzyme also exhibited 1,4-β-mannosidase and cellodextrinase activities. Man5A-TM1, compared to either Man5A-TM2 or Man5B, had higher catalytic activity with soluble and insoluble polysaccharides, indicating that the CBMs enhance catalysis of Man5A. Furthermore, Man5A-TM1 acted synergistically with Man5B in the hydrolysis of β-mannan and carboxymethyl cellulose. The versatility of the two enzymes, therefore, makes them a resource for depolymerization of mannan-containing polysaccharides in the biofuel industry. Furthermore, on the basis of the biochemical and genomic data, a molecular mechanism for utilization of mannan-containing nutrients by C. polysaccharolyticus is proposed.

scholarly journals Unravelling the diversity of glycoside hydrolase family 13 α-amylases from Lactobacillus plantarum WCFS1

2019 ◽  
Vol 18 (1) ◽  
Author(s):  
Laura Plaza-Vinuesa ◽  
Oswaldo Hernandez-Hernandez ◽  
F. Javier Moreno ◽  
Blanca de las Rivas ◽  
Rosario Muñoz
Keyword(s):  
Lactobacillus Plantarum ◽  
Glycoside Hydrolase ◽  
Catalytic Efficiency ◽  
Hydrolytic Activity ◽  
Structural Features ◽  
Biochemical Properties ◽  
Glycoside Hydrolase Family ◽  
Reaction Conditions ◽  
Genes Encoding ◽  
Hydrolysis Of

Abstract Background α-Amylases specifically catalyse the hydrolysis of the internal α-1, 4-glucosidic linkages of starch. Glycoside hydrolase (GH) family 13 is the main α-amylase family in the carbohydrate-active database. Lactobacillus plantarum WCFS1 possesses eleven proteins included in GH13 family. Among these, proteins annotated as maltose-forming α-amylase (Lp_0179) and maltogenic α-amylase (Lp_2757) were included. Results In this study, Lp_0179 and Lp_2757 L. plantarum α-amylases were structurally and biochemically characterized. Lp_2757 displayed structural features typical of GH13_20 subfamily which were absent in Lp_0179. Genes encoding Lp_0179 (Amy2) and Lp_2757 were cloned and overexpressed in Escherichia coli BL21(DE3). Purified proteins showed high hydrolytic activity on pNP-α-D-maltopyranoside, being the catalytic efficiency of Lp_0179 remarkably higher. In relation to the hydrolysis of starch-related carbohydrates, Lp_0179 only hydrolysed maltopentaose and dextrin, demonstrating that is an exotype glucan hydrolase. However, Lp_2757 was also able to hydrolyze cyclodextrins and other non-cyclic oligo- and polysaccharides, revealing a great preference towards α-1,4-linkages typical of maltogenic amylases. Conclusions The substrate range as well as the biochemical properties exhibited by Lp_2757 maltogenic α-amylase suggest that this enzyme could be a very promising enzyme for the hydrolysis of α-1,4 glycosidic linkages present in a broad number of starch-carbohydrates, as well as for the investigation of an hypothetical transglucosylation activity under appropriate reaction conditions.

scholarly journals Biochemical properties of free and immobilized Candida viswanathii lipase on octyl-agarose support: Hydrolysis of triacylglycerol and soy lecithin

2018 ◽  
Vol 65 ◽  
pp. 71-80 ◽  
Author(s):  
Alex Fernando de Almeida ◽  
César Rafael Fanchini Terrasan ◽  
Cárol Cabral Terrone ◽  
Sâmia Maria Tauk-Tornisielo ◽  
Eleonora Cano Carmona
Keyword(s):  
Biochemical Properties ◽  
Soy Lecithin ◽  
Candida Viswanathii ◽  
Hydrolysis Of

scholarly journals A novel plant enzyme with dual activity: an atypical Nudix hydrolase and a dipeptidyl peptidase III

2017 ◽  
Vol 398 (1) ◽  
pp. 101-112 ◽  
Author(s):  
Zrinka Karačić ◽  
Bojana Vukelić ◽  
Gabrielle H. Ho ◽  
Iva Jozić ◽  
Iva Sučec ◽  
...  
Keyword(s):  
Physcomitrella Patens ◽  
Catalytic Efficiency ◽  
Peptide Bond ◽  
Dipeptidyl Peptidase ◽  
Biochemical Properties ◽  
Binding Motif ◽  
Nudix Hydrolase ◽  
Purine Nucleotides ◽  
Uncharacterized Protein ◽  
Hydrolysis Of

Abstract In a search for plant homologues of dipeptidyl peptidase III (DPP III) family, we found a predicted protein from the moss Physcomitrella patens (UniProt entry: A9TLP4), which shared 61% sequence identity with the Arabidopsis thaliana uncharacterized protein, designated Nudix hydrolase 3. Both proteins contained all conserved regions of the DPP III family, but instead of the characteristic hexapeptide HEXXGH zinc-binding motif, they possessed a pentapeptide HEXXH, and at the N-terminus, a Nudix box, a hallmark of Nudix hydrolases, known to act upon a variety of nucleoside diphosphate derivatives. To investigate their biochemical properties, we expressed heterologously and purified Physcomitrella (PpND) and Arabidopsis (AtND) protein. Both hydrolyzed, with comparable catalytic efficiency, the isopentenyl diphosphate (IPP), a universal precursor for the biosynthesis of isoprenoid compounds. In addition, PpND dephosphorylated four purine nucleotides (ADP, dGDP, dGTP, and 8-oxo-dATP) with strong preference for oxidized dATP. Furthermore, PpND and AtND showed DPP III activity against dipeptidyl-2-arylamide substrates, which they cleaved with different specificity. This is the first report of a dual activity enzyme, highly conserved in land plants, which catalyzes the hydrolysis of a peptide bond and of a phosphate bond, acting both as a dipeptidyl peptidase III and an atypical Nudix hydrolase.

scholarly journals Depolymerization of β-1,6-N-Acetyl-d-Glucosamine Disrupts the Integrity of Diverse Bacterial Biofilms

2005 ◽  
Vol 187 (1) ◽  
pp. 382-387 ◽  
Author(s):  
Yoshikane Itoh ◽  
Xin Wang ◽  
B. Joseph Hinnebusch ◽  
James F. Preston ◽  
Tony Romeo
Keyword(s):  
Mass Spectrometry ◽  
Escherichia Coli ◽  
Enzymatic Hydrolysis ◽  
Pseudomonas Fluorescens ◽  
Yersinia Pestis ◽  
Staphylococcus Epidermidis ◽  
Biofilm Formation ◽  
Bacterial Biofilms ◽  
First Time ◽  
Hydrolysis Of

ABSTRACT Polymeric β-1,6-N-acetyl-d-glucosamine (poly-β-1,6-GlcNAc) has been implicated as an Escherichia coli and Staphylococcus epidermidis biofilm adhesin, the formation of which requires the pgaABCD and icaABCD loci, respectively. Enzymatic hydrolysis of poly-β-1,6-GlcNAc, demonstrated for the first time by chromatography and mass spectrometry, disrupts biofilm formation by these species and by Yersinia pestis and Pseudomonas fluorescens, which possess pgaABCD homologues.

Characterization of Protease from the Myxosporean Salmon Parasite, Henneguya salminicola

1984 ◽  
Vol 41 (2) ◽  
pp. 371-376 ◽  
Author(s):  
E. Bilinski ◽  
N. P. Boyce ◽  
R. E. E. Jonas ◽  
M. D. Peters
Keyword(s):  
Metal Ions ◽  
Sockeye Salmon ◽  
Frozen Storage ◽  
Sulfhydryl Group ◽  
Maximal Activity ◽  
Biochemical Properties ◽  
Divalent Metal Ions ◽  
Myxosporean Parasite ◽  
Preliminary Activation ◽  
Hydrolysis Of

The biochemical properties of the protease(s) from the myxosporean parasite, Henneguya salminicola, occurring in the muscle of sockeye salmon, Oncorhynchus nerka, were examined. The cysts of the parasite contained a soluble, heat-labile protease, having maximal activity at pH 3.0 for hydrolysis of hemoglobin and 4.5 for hydrolysis of salmon muscle proteins. The inhibitions obtained from metal ions and from sulfhydryl group binding reagents suggest the involvement of sulfhydryl groups in the expression of enzyme activity. The enzyme required no preliminary activation by disulfide bond reducing agents and it had no obvious divalent metal ions requirement. The hydrolysis of salmon muscle by H. salminicola protease was strongly temperature dependent. Although proteolysis occurred at the highest rate at 30–40 °C, the enzyme remained active in iced muscle. The proteolytic activity was arrested completely in frozen muscle but the enzyme was only slightly inactivated by a prolonged frozen storage (−28 °C). The presence of the parasite had no marked effect on the texture of raw or cooked flesh.

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