scholarly journals Extraction and Characterization of Pepsin Enzyme from Tuna (Thunnus albacares) Gastric

2018 ◽  
Vol 21 (3) ◽  
pp. 486 ◽  
Author(s):  
Ewi Pasaribu ◽  
Tati Nurhayati ◽  
Mala Nurilmala
Keyword(s):  
Crude Extract ◽  
Metal Ion ◽  
Specific Activity ◽  
Gastric Wall ◽  
Extraction Process ◽  
Thunnus Albacares ◽  
Fractional Precipitation ◽  
Working Temperature ◽  
Ph Range

Fish gastric is a by-product of fishery waste that has the potential to be developed as a source of pepsin enzyme, such as tuna gastric. The purpose of this study was to extract pepsin from tuna gastric and characterize the enzyme pepsin after dialysis step. The extraction process of pepsin carried out by separating the gastric wall fluid by adding Tris-HCl buffer pH 7.5, then proceed by ammonium sulphate (NH4)2SO4 fractional precipitation from 20% to 80% followed by dialysis. The results showed that the crude extract of the pepsin enzyme had a specific activity of 0.251 mg/mL. Pepsin extract from precipitation with 30-40% fraction had 4,274 U/mg activity and after dialysis, pepsin had 5,137 U / mg activity. The pepsin obtained from gastric tuna had a working temperature in the range of 20-60°C and the working pH is in the pH range 2-3.5. The metal ion, namely FeCl3 and ZnCl2 increased the activity of the pepsin by 2.97 times and 1.92 times.

scholarly journals Production and Characterization of Esterase in Lantinus tigrinus for Degradation of Polystyrene

2013 ◽  
Vol 62 (1) ◽  
pp. 101-108 ◽  
Author(s):  
LUBNA TAHIR ◽  
MUHAMMAD ISHTIAQ ALI ◽  
MUHAMMAD ZIA ◽  
NAIMA ATIQ ◽  
FARIHA HASAN ◽  
...  
Keyword(s):  
Acidic Medium ◽  
Specific Activity ◽  
Mineral Salt Medium ◽  
Tween 80 ◽  
Synthetic Polymers ◽  
Ftir Analysis ◽  
Polystyrene Film ◽  
Biological Degradation ◽  
Ph Range

Polystyrene is considered stable to biological degradation. Lantinus tigrinus isolated from wood sample produced esterase in growth medium under normal conditions. However, acidic medium, 37 degrees C temperature, presence of tween 80; and urea and yeast extract in mineral salt medium enhance the production of esterase and specific activity. Purified esterase was active at broad pH range and 45 degrees C. FTIR analysis confirmed that esterase produced by Lantinus tigrinus effectively degraded polystyrene film and broke macromolecules down to non-toxic molecules. This study concludes that the presence of Lantinus tigrinus at dumping sites can be exploited for waste management containing high molecular weight synthetic polymers.

scholarly journals Characterization of a Robust and pH-Stable Tannase from Mangrove-Derived Yeast Rhodosporidium diobovatum Q95

Marine Drugs ◽  
2020 ◽  
Vol 18 (11) ◽  
pp. 546
Author(s):  
Jie Pan ◽  
Ni-Na Wang ◽  
Xue-Jing Yin ◽  
Xiao-Ling Liang ◽  
Zhi-Peng Wang
Keyword(s):  
Gallic Acid ◽  
Tannic Acid ◽  
Specific Activity ◽  
Maximum Activity ◽  
Sds Page ◽  
Rhodosporidium Diobovatum ◽  
Bacteria And Fungi ◽  
Ph Range ◽  
First Time

Tannase plays a crucial role in many fields, such as the pharmaceutical industry, beverage processing, and brewing. Although many tannases derived from bacteria and fungi have been thoroughly studied, those with good pH stabilities are still less reported. In this work, a mangrove-derived yeast strain Rhodosporidium diobovatum Q95, capable of efficiently degrading tannin, was screened to induce tannase, which exhibited an activity of up to 26.4 U/mL after 48 h cultivation in the presence of 15 g/L tannic acid. The tannase coding gene TANRD was cloned and expressed in Yarrowia lipolytica. The activity of recombinant tannase (named TanRd) was as high as 27.3 U/mL. TanRd was purified by chromatography and analysed by SDS-PAGE, showing a molecular weight of 75.1 kDa. The specific activity of TanRd towards tannic acid was 676.4 U/mg. Its highest activity was obtained at 40 °C, with more than 70% of the activity observed at 25–60 °C. Furthermore, it possessed at least 60% of the activity in a broad pH range of 2.5–6.5. Notably, TanRd was excellently stable at a pH range from 3.0 to 8.0; over 65% of its maximum activity remained after incubation. Besides, the broad substrate specificity of TanRd to esters of gallic acid has attracted wide attention. In view of the above, tannase resources were developed from mangrove-derived yeasts for the first time in this study. This tannase can become a promising material in tannin biodegradation and gallic acid production.

scholarly journals Characterization of a New Intracellular Alginate Lyase with Metal Ions-Tolerant and pH-Stable Properties

Marine Drugs ◽  
2020 ◽  
Vol 18 (8) ◽  
pp. 416
Author(s):  
Yan Ma ◽  
Jie Li ◽  
Xin-Yue Zhang ◽  
Hao-Dong Ni ◽  
Feng-Biao Wang ◽  
...  
Keyword(s):  
Metal Ion ◽  
Brown Seaweed ◽  
Alginate Lyase ◽  
Industrial Applications ◽  
Alginate Oligosaccharides ◽  
Potential Applications ◽  
Pharmaceutical Industries ◽  
Ph Range ◽  
Alginate Lyases

Alginate lyases play an important role in alginate oligosaccharides (AOS) preparation and brown seaweed processing. Many extracellular alginate lyases have been characterized to develop efficient degradation tools needed for industrial applications. However, few studies focusing on intracellular alginate lyases have been conducted. In this work, a novel intracellular alkaline alginate lyase Alyw202 from Vibrio sp. W2 was cloned, expressed and characterized. Secretory expression was performed in a food-grade host, Yarrowia lipolytica. Recombinant Alyw202 with a molecular weight of approximately 38.3 kDa exhibited the highest activity at 45 °C and more than 60% of the activity in a broad pH range of 3.0 to 10.0. Furthermore, Alyw202 showed remarkable metal ion-tolerance, NaCl independence and the capacity of degrading alginate into oligosaccharides of DP2-DP4. Due to the unique pH-stable and high salt-tolerant properties, Alyw202 has potential applications in the food and pharmaceutical industries.

scholarly journals Investigation of the first step of biotin biosynthesis in Bacillus sphaericus. Purification and characterization of the pimeloyl-CoA synthase, and uptake of pimelate

1992 ◽  
Vol 287 (3) ◽  
pp. 685-690 ◽  
Author(s):  
O Ploux ◽  
P Soularue ◽  
A Marquet ◽  
R Gloeckler ◽  
Y Lemoine
Keyword(s):  
Metal Ion ◽  
Specific Activity ◽  
Bacillus Sphaericus ◽  
Careful Study ◽  
Metal Chelators ◽  
Ph Profile ◽  
E Coli ◽  
Coa Ligase ◽  
Hydrolysis Of

The pimeloyl-CoA synthase from Bacillus sphaericus has been purified to homogeneity from an overproducing strain of Escherichia coli. The purification yielded milligram quantities of the synthase with a specific activity of 1 unit/mg of protein. Analysis of the products showed that this enzyme catalysed the transformation of pimelate into pimeloyl-CoA with concomitant hydrolysis of ATP to AMP. Using a continuous spectrophotometric assay, we have examined the catalytic properties of the pure enzyme. The pH profile under Vmax. conditions showed a maximum around 8.5. Apparent Km values for pimelate, CoASH, ATP. Mg2- and Mg2+ were respectively 145 microM, 33 microM, 170 microM and 2.3 mM. The enzyme was inhibited by Mg2+ above 10 mM. This acid-CoA ligase exhibited a very sharp substrate specificity, e.g. neither GTP nor pimelate analogues (di- or mono-carboxylic acids) were processed. The bivalent metal ion requirement was also investigated: Mn2+ (73%) and Co2+ (32%) but not Ca2+ could replace Mg2+. The enzyme was inhibited by metal chelators such as 1,10-phenanthroline and EDTA. The synthase was a homodimer with a 28,000-M(r) subunit. N-Terminal sequencing definitely proved that this enzyme was encoded by the bioW gene. A careful study of pimelate uptake by B. sphaericus, E. coli and Pseudomonas dentrificans showed that this metabolite crossed the membrane of these microorganisms by passive diffusion, ruling out the involvement of the bioX gene product as pimelate carrier.

Characterization of Protein Isolates from Defatted Chinese Hickory (Carya Cathayensis Sarg) Kernels

2011 ◽  
Vol 343-344 ◽  
pp. 368-373
Author(s):  
Zhi Ping He ◽  
Lin Chun Mao ◽  
Cun Shan Zhou ◽  
Feng Hua Wu
Keyword(s):  
Dry Weight ◽  
Extraction Process ◽  
Protein Isolate ◽  
Protein Nitrogen ◽  
Gelation Concentration ◽  
Carya Cathayensis ◽  
Ph Range ◽  
Emulsifying Capacity ◽  
Carya Cathayensis Sarg

Chinese hickory (Carya cathayensis Sarg.) kernel protein isolate (CKPI) was isolated from hinese hickory kernel defatted flour (CKDF) by isoelectric precipitation. CKPI was evaluated for chemical composition and selected functional properties. CKPI contained over 72.0% dry weight (DW) of protein. Nitrogen solubility curves for CKPI were pH-dependent in the pH range of 2.0-12.0 with minimum solubility observed at pH 4.0 (9.8%) and maximum solubility at pH 12.0 (89.9%). Minimum emulsifying capacity (EC) and emulsifying stability (ES) of CKPI were observed at pH 4.0. CKPI had a least gelation concentration of (LGC) of 6% (w/v) at pH 4.0. Results indicated that Chinese hickory kernel may be a new protein source with huge exploitation potential after oil extraction process in China.

scholarly journals Purification, Characterization and Degradation Performance of a Novel Dextranase from Penicillium cyclopium CICC-4022

2019 ◽  
Vol 20 (6) ◽  
pp. 1360 ◽  
Author(s):  
Ruijie Huang ◽  
Lei Zhong ◽  
Fengwei Xie ◽  
Liming Wei ◽  
Lanfang Gan ◽  
...  
Keyword(s):  
Molecular Mass ◽  
Metal Ion ◽  
Specific Activity ◽  
Gel Filtration ◽  
Gel Permeation Chromatography ◽  
Fractional Precipitation ◽  
Penicillium Species ◽  
Average Molecular Mass ◽  
Penicillium Cyclopium ◽  
Effects Of Temperature

A novel dextranase was purified from Penicillium cyclopium CICC-4022 by ammonium sulfate fractional precipitation and gel filtration chromatography. The effects of temperature, pH and some metal ions and chemicals on dextranase activity were investigated. Subsequently, the dextranase was used to produce dextran with specific molecular mass. Weight-average molecular mass (Mw) and the ratio of weight-average molecular mass/number-average molecular mass, or polydispersity index (Mw/Mn), of dextran were measured by multiple-angle laser light scattering (MALS) combined with gel permeation chromatography (GPC). The dextranase was purified to 16.09-fold concentration; the recovery rate was 29.17%; and the specific activity reached 350.29 U/mg. Mw of the dextranase was 66 kDa, which is similar to dextranase obtained from other Penicillium species reported previously. The highest activity was observed at 55 °C and a pH of 5.0. This dextranase was identified as an endodextranase, which specifically degraded the α-1,6 glucosidic bonds of dextran. According to metal ion dependency tests, Li+, Na+ and Fe2+ were observed to effectively improve the enzymatic activity. In particular, Li+ could improve the activity to 116.28%. Furthermore, the dextranase was efficient at degrading dextran and the degradation rate can be well controlled by the dextranase activity, substrate concentration and reaction time. Thus, our results demonstrate the high potential of this dextranase from Penicillium cyclopium CICC-4022 as an efficient enzyme to produce specific clinical dextrans.

scholarly journals Characterization of an Alkaline Alginate Lyase with pH-Stable and Thermo-Tolerance Property

Marine Drugs ◽  
2019 ◽  
Vol 17 (5) ◽  
pp. 308 ◽  
Author(s):  
Yanan Wang ◽  
Xuehong Chen ◽  
Xiaolin Bi ◽  
Yining Ren ◽  
Qi Han ◽  
...  
Keyword(s):  
Marine Bacterium ◽  
Specific Activity ◽  
Alginate Lyase ◽  
Initial Activity ◽  
Alginate Oligosaccharides ◽  
Wide Ph Range ◽  
Encoding Gene ◽  
Ph Range ◽  
Alginate Lyases

Alginate oligosaccharides (AOS) show versatile bioactivities. Although various alginate lyases have been characterized, enzymes with special characteristics are still rare. In this study, a polysaccharide lyase family 7 (PL7) alginate lyase-encoding gene, aly08, was cloned from the marine bacterium Vibrio sp. SY01 and expressed in Escherichia coli. The purified alginate lyase Aly08, with a molecular weight of 35 kDa, showed a specific activity of 841 U/mg at its optimal pH (pH 8.35) and temperature (45 °C). Aly08 showed good pH-stability, as it remained more than 80% of its initial activity in a wide pH range (4.0–10.0). Aly08 was also a thermo-tolerant enzyme that recovered 70.8% of its initial activity following heat shock treatment for 5 min. This study also demonstrated that Aly08 is a polyG-preferred enzyme. Furthermore, Aly08 degraded alginates into disaccharides and trisaccharides in an endo-manner. Its thermo-tolerance and pH-stable properties make Aly08 a good candidate for further applications.

scholarly journals Purification and Characterization of a Novel Thermostable α-l-Arabinofuranosidase from a Color-Variant Strain of Aureobasidium pullulans

1998 ◽  
Vol 64 (1) ◽  
pp. 216-220 ◽  
Author(s):  
Badal C. Saha ◽  
Rodney J. Bothast
Keyword(s):  
Column Chromatography ◽  
Metal Ion ◽  
Aureobasidium Pullulans ◽  
Specific Activity ◽  
Gel Filtration ◽  
Variant Strain ◽  
Color Variant ◽  
C 50 ◽  
Hydrolysis Of

ABSTRACT A color-variant strain of Aureobasidium pullulans (NRRL Y-12974) produced α-l-arabinofuranosidase (α-l-AFase) when grown in liquid culture on oat spelt xylan. An extracellular α-l-AFase was purified 215-fold to homogeneity from the culture supernatant by ammonium sulfate treatment, DEAE Bio-Gel A agarose column chromatography, gel filtration on a Bio-Gel A-0.5m column, arabinan-Sepharose 6B affinity chromatography, and SP-Sephadex C-50 column chromatography. The purified enzyme had a native molecular weight of 210,000 and was composed of two equal subunits. It had a half-life of 8 h at 75°C, displayed optimal activity at 75°C and pH 4.0 to 4.5, and had a specific activity of 21.48 μmol · min−1· mg−1 of protein againstp-nitrophenyl-α-l-arabinofuranoside (pNPαAF). The purified α-l-AFase readily hydrolyzed arabinan and debranched arabinan and released arabinose from arabinoxylans but was inactive against arabinogalactan. TheKm values of the enzyme for the hydrolysis of pNPαAF, arabinan, and debranched arabinan at 75°C and pH 4.5 were 0.26 mM, 2.14 mg/ml, and 3.25 mg/ml, respectively. The α-l-AFase activity was not inhibited at all byl-arabinose (1.2 M). The enzyme did not require a metal ion for activity, and its activity was not affected byp-chloromercuribenzoate (0.2 mM), EDTA (10 mM), or dithiothreitol (10 mM).

scholarly journals Characterization of a Novel Zinc-Containing, Lysine-Specific Aminopeptidase from the Hyperthermophilic Archaeon Pyrococcus furiosus

2005 ◽  
Vol 187 (6) ◽  
pp. 2077-2083 ◽  
Author(s):  
Sherry V. Story ◽  
Claudia Shah ◽  
Francis E. Jenney ◽  
Michael W. W. Adams
Keyword(s):  
Metal Ion ◽  
Specific Activity ◽  
Pyrococcus Furiosus ◽  
High Specific Activity ◽  
Native Form ◽  
Genome Database ◽  
Hyperthermophilic Archaeon ◽  
Cell Extracts ◽  
Hydrolysis Of

ABSTRACT Cell extracts of the proteolytic, hyperthermophilic archaeon Pyrococcus furiosus contain high specific activity (11 U/mg) of lysine aminopeptidase (KAP), as measured by the hydrolysis of l-lysyl-p-nitroanilide (Lys-pNA). The enzyme was purified by multistep chromatography. KAP is a homotetramer (38.2 kDa per subunit) and, as purified, contains 2.0 ± 0.48 zinc atoms per subunit. Surprisingly, its activity was stimulated fourfold by the addition of Co2+ ions (0.2 mM). Optimal KAP activity with Lys-pNA as the substrate occurred at pH 8.0 and a temperature of 100°C. The enzyme had a narrow substrate specificity with di-, tri-, and tetrapeptides, and it hydrolyzed only basic N-terminal residues at high rates. Mass spectroscopy analysis of the purified enzyme was used to identify, in the P. furiosus genome database, a gene (PF1861) that encodes a product corresponding to 346 amino acids. The recombinant protein containing a polyhistidine tag at the N terminus was produced in Escherichia coli and purified using affinity chromatography. Its properties, including molecular mass, metal ion dependence, and pH and temperature optima for catalysis, were indistinguishable from those of the native form, although the thermostability of the recombinant form was dramatically lower than that of the native enzyme (half-life of approximately 6 h at 100°C). Based on its amino acid sequence, KAP is part of the M18 family of peptidases and represents the first prokaryotic member of this family. KAP is also the first lysine-specific aminopeptidase to be purified from an archaeon.

Poly[1,3(2-ethoxy-4-hydroxy-5-propylphenylene)propylene] copolymer as an ion exchanger

1994 ◽  
Vol 6 (3) ◽  
pp. 201-208 ◽  
Author(s):  
Jayesh R Patel ◽  
Dipan H Sutaria ◽  
Magan N Patel
Keyword(s):  
Metal Ion ◽  
Metal Uptake ◽  
Polyphosphoric Acid ◽  
Average Molecular Weight ◽  
Acid Catalyst ◽  
Wide Ph Range ◽  
Metal Ion Uptake ◽  
Viscosity Study ◽  
Ph Range

The present paper reports the synthesis and characterization of a copolymer, The copolymer sample was synthesized by Friedel-Crafts polycondensation of 2-hydroxy-4 ethoxypropiophenone with 1,3-propane diol in the presence of polyphosphoric acid catalyst. The copolymer was characterized by elemental analysis, IR spectroscopy and viscosity study, and its number-average molecular weight (M) was determined by non-aqueous titration and found to be 2855 g mol- 1. Chelation ion-exchange properties have also been studied, employing a batch-equilibration method. It was employed to study selectivity of metal ion uptake over a wide pH range and in media of various ionic strengths. The overall rate of metal uptake follows the order UOl + > Fe3 + > Cu2 + > Ni2 +.

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