scholarly journals Molecular Identification of Indigenous Lactic Acid Bacteria, Partial Purification and Characterization of β-Galactosidase Produced

2017 ◽  
Vol 3 (4) ◽  
pp. 247
Author(s):  
Tatik Khusniati ◽  
Sulistiania . ◽  
Bellen Nastitie Pamela Fury ◽  
Syamsul Falah ◽  
Abdul Choliq
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Plantarum ◽  
Ammonium Sulphate ◽  
Molecular Identification ◽  
Specific Activity ◽  
Partial Purification ◽  
Optimum Temperature ◽  
Purification And Characterization

<p>β-Galactosidase is enzyme which hydrolyze lactose to glucose and galactose, as lactose hydrolyzer. To know indigenous lactic acid bacteria (LAB) characteristics, indigenous LAB identification, partial purification and characterization of β-galactosidase produced was researched. LAB was molecularly identified, β-galactosidase partial purification was conducted by precipitation followed dialysis. β-Galactosidase  characterization was based on optimum activities of pH and temperature. The results show that LAB was identified as <em>Lactobacillus plantarum</em> B123. Optimum activity of precipited β-galactosidase was reached at 50 % ammonium sulphate. Activity and specific activity of 50 % precipited β-galactosidase were 95.675 U · mg<sup>–1</sup> and 32.268 U · mg<sup>–1</sup>, respectively. Precipited β-galactosidase resulted a purification level of 3.99 fold, a yield of 38.73 %, and a specific activity of 32.27 U · mg<sup>–1</sup> protein, while dialyzed β-galactosidase resulted 7.61 fold, 10.67 %, and 61.53 U· mg<sup>–1</sup> protein. Optimum temperature and pH for crude β-galactosidase were found at 55 °C and 7.0, respectively, while that dialyzed β-galactosidase were optimized at 50 °C and 7.0. Based on partial purification and characterization, <em>Lactobacillus plantarum</em> B123 is indigenous LAB which good for production of β-galactosidase.</p><div><p class="Els-keywords"><em> </em></p><p class="Els-keywords"><strong>Keywords:</strong> characterization; dialysis; lactic acid bacteria; <em>Lactobacillus plantarum</em> B 123; β-galactosidase.</p></div>

scholarly journals PURIFIKASI PARSIAL DAN KARAKTERISASI ß-GALAKTOSIDASE Lactobacillus plantarum B123 INDIGENOS DAN HIDROLISIS LAKTOSA UNTUK PRODUKSI SUSU ULTRA HIGH TEMPERATURE RENDAH LAKTOSA

2015 ◽  
Vol 17 (2) ◽  
pp. 147-161
Author(s):  
Tatik Khusniati ◽  
Neny Mariyani ◽  
Hanifah Nuryani Lioe ◽  
Didah Nur Faridah ◽  
Abdul Choliq ◽  
...  
Keyword(s):  
High Temperature ◽  
Lactobacillus Plantarum ◽  
Lactose Intolerance ◽  
Specific Activity ◽  
Partial Purification ◽  
Lactose Hydrolysis ◽  
Galactosidase Activity ◽  
Purification And Characterization ◽  
Uht Milk

β-Galactosidase is enzyme which hidrolyze lactose to glucose and galactose. This enzyme is used in production low lactose milk for consumption human which have lactose intolerance. Partial purification of β-galactosidase is important to be conducted to increase  β-galactosidase activity in order to its hydrolysis potency on UHT milk lactose increased.This research was aimed to production by partially purification and characterization indigenous β-galactosidase from Lactobacillus plantarum B123, and lactose hydrolysis for production low lactose UHT milk. Partially purification were precipitation following dialysis. Characterization included optimazion and stabilization of enzyme, while lactose hydrolisis for production low lactose UHT milk was detected by enzymatic GOD-POD kit. The results showed that production of β-galactosidase by using partial purification increased from 21.51 ± 0.23 U/mL (crude) to 106.34 ± 0.56 U/mL (dialysis).  The optimum crude β-galactosidase activity was reached in precipitation by using 60 % ammonium sulphate.  The purity of crude β-galactosidase increased 3.71 times after precipitation, and 14.28  times  after dialysis. Characterization of β-galactosidase showed that  optimum activities of crude and dialyzed β-galactosidase were at pH 6.5 and 50 oC, respectively. Stability of crude β-galactosidase incubated for 1 h were at pH: 5.0-8.5 and 25-50 °C. Specific activity of crude β-galactosidase was 15.05 U/mg protein, while that dialyzed β-galactosidase was 109.58 U/mg protein. Lactose hidrolysis to produce low lactose UHT milk showed that glucose concentration increased with the increase of hidrolysis time. Time needed to hidrolyze lactose 50 % with 4.8 U/mL β-galactosidase at 50°C was 6.08 h. In conclusion that indigenous β-galactosidase from Lactobacillus plantarum B123 purified partially can be used as lactose hidrolyzer in production of low lactose UHT milk.Key words : b-galactosidase, indigenous Lactobacillus plantarum B123, purification, lactose hidrolysis, UHT Milk

scholarly journals Partial purification and characterization of human erythrocyte phosphoglycollate phosphatase

1980 ◽  
Vol 191 (1) ◽  
pp. 117-124 ◽  
Author(s):  
R Zecher ◽  
H U Wolf
Keyword(s):  
Specific Activity ◽  
Total Activity ◽  
Partial Purification ◽  
Half Maximum ◽  
Purification And Characterization ◽  
Dimeric Molecule ◽  
Maximum Inhibition ◽  
Optimum Ph ◽  
Triton X

Human erythrocytes contain a phosphatase that is highly specific for phosphoglycollate. It shows optimum pH of 6.7 and has Km 1 mM for phosphoglycollate. The molecular weight appears to be about 72000. The enzyme is a dimeric molecule having subunits of mol. wt. about 35000. It could be purified approx. 4000-fold up to a specific activity of 5.98 units/mg of protein. The activity of the enzyme is Mg2+-dependent. Co2+, and to a smaller extent Mn2+, may substitute for Mg2+. Half-maximum inhibition of the phosphatase by 5,5′-dithiobis-(2-nitrobenzoate), EDTA and NaF is obtained at 0.5 microM, 1 mM and 4 mM respectively. Moreover, it needs a univalent cation for optimum activity. Phosphoglycollate phosphatase is a cytoplasmic enzyme. Approx. 5% of its total activity is membrane-associated. This part of activity can be approx. 70% solubilized by freezing, thawing and treatment with 0.25% Triton X-100.

scholarly journals Purification and characterization of alkaline phosphatase from lactic acid bacteria

RSC Advances ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 354-360 ◽  
Author(s):  
Yu-Hao Chu ◽  
Xin-Xin Yu ◽  
Xing Jin ◽  
Yu-Tang Wang ◽  
Duo-Jia Zhao ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Organophosphorus Pesticides ◽  
Purification And Characterization

Alkaline phosphatase (ALP) excreted from lactic acid bacteria (LAB) showed the ability to degrade organophosphorus pesticides.

Catechol 1,2-dioxygenase from α-naphthol degrading thermophilic Geobacillus sp. strain: purification and properties

2009 ◽  
Vol 4 (1) ◽  
pp. 68-73 ◽  
Author(s):  
Gražina Giedraityte ◽  
Lilija Kalėdienė
Keyword(s):  
Specific Activity ◽  
Relative Molecular Mass ◽  
Gel Chromatography ◽  
Optimum Temperature ◽  
Ammonium Sulfate Precipitation ◽  
Purification And Characterization ◽  
Purification And Properties ◽  
Catechol 1,2 Dioxygenase ◽  
Temperature Optima

AbstractThe purpose of this study was purification and characterization of catechol 1,2-dioxygenase from Geobacillus sp. G27 strain, which degrades α-naphthol by the β-ketoadipate pathway. The catechol 1,2-dioxygenase (C1,2O) was purified using four steps of ammonium sulfate precipitation, DEAE-celullose, Sephadex G-150 and hydroxylapatite chromatographies. The enzyme was purified about 18-fold with a specific activity of 7.42 U mg of protein−1. The relative molecular mass of the native enzyme estimated on gel chromatography of Sephadex G-150 was 96 kDa. The pH and temperature optima for enzyme activity were 7 and 60°C, respectively. A half-life of the catechol 1,2-dioxygenase at the optimum temperature was 40 min. The kinetic parameters of the Geobacillus sp. G27 strain catechol 1,2-dioxygenase were determined. The enzyme had apparent Km of 29 µM for catechol and the cleavage activities for methylcatechols were much less than for catechol and no activity with gentisate or protocatechuate was detected.

scholarly journals Characterization of non-starter lactic acid bacteria in traditionally produced home-made Radan cheese during ripening

2011 ◽  
Vol 63 (1) ◽  
pp. 1-10 ◽  
Author(s):  
Natasa Jokovic ◽  
Maja Vukasinovic ◽  
Katarina Veljovic ◽  
Maja Tolinacki ◽  
L. Topisirovic
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Plantarum ◽  
Enterococcus Faecium ◽  
Leuconostoc Mesenteroides ◽  
Streptococcus Thermophilus ◽  
Predominant Species ◽  
Lactobacillus Paraplantarum ◽  
Good Agreement

Two hundred thirteen non-starter lactic acid bacteria isolated from Radan cheese during ripening were identified with both a classical biochemical test and rep-PCR with (GTG)5 primer. For most isolates, which belong to the Lactococcus lactis subsp. lactis, Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus paraplantarum and Enterococcus faecium, a phenotypic identification was in good agreement with rep-PCR identification. Lactococeus lactis subsp. lactis, Enterococcus faecium and subspecies from the Lenconostoc mesenteroides group were the dominant population of lactic acid bacteria in cheese until 10 days of ripening and only one Streptococcus thermophilus strain was isolated from the 5-day-old cheese sample. As ripening progressed, Lactobacillus plantarum became the predominant species together with the group of heterofermentative species of lactobacilli that could not be precisely identified with rep-PCR.

Post-proline endopeptidase. Partial purification and characterization of the enzyme from pig kidneys

1982 ◽  
Vol 47 (4) ◽  
pp. 1139-1148 ◽  
Author(s):  
Karel Hauzer ◽  
Linda Servítová ◽  
Tomislav Barth ◽  
Karel Jošt
Keyword(s):  
Specific Activity ◽  
Gel Filtration ◽  
Peptide Bond ◽  
Exchange Chromatography ◽  
Partial Purification ◽  
Purification And Characterization ◽  
Optimum Ph ◽  
Hydrolysis Of ◽  
Prolyl Leucyl Glycinamide

Post-proline endopeptidase was isolated from pig kidneys and partially purified. The procedure consisted of fractionation with ammonium sulphate, ion exchange chromatography on DEAE-Sephadex A-50, gel filtration on Sephadex G-200 and rechromatography on DEAE-Sephadex A-50. The preparation had 55 times higher specific activity than the crude extract and did not contain any contaminating enzymic activities. The enzyme cleaved a number of proline-containing peptides and was strictly specific in catalyzing the hydrolysis of the peptide bond on the carboxyl side of the proline residue. The optimum pH for the hydrolysis of the synthetic peptides benzyl-oxycarbonylglycyl-prolyl-leucyl-glycinamide and benzyloxycarbonyl-glycyl-proline β-naphtylamide was 7.8-8.0 and, in the case of benzyloxycarbonylglycyl-proline p-nitroanilide, 7.2 to 7.5. For the hydrolysis of the tetrapeptide benzyloxycarbonylglycyl-prolyl-leucyl-glycinamide, the Km value of 75 μ mol l-1 was obtained.

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