Adenosine Aminohydrolase from Halobacterium cutirubrum

1973 ◽  
Vol 51 (5) ◽  
pp. 621-626 ◽  
Author(s):  
Randy J. Bauer ◽  
David M. Carlberg
Keyword(s):  
Ammonium Sulfate ◽  
Gel Filtration ◽  
Maximal Activity ◽  
Halophilic Bacterium ◽  
Ammonium Sulfate Precipitation ◽  
Two Phase ◽  
Liquid Polymer ◽  
Optimum Ph ◽  
Pyrimidine Bases

The extreme halophilic bacterium Halobacterium cutirubrum was examined for base, nucleoside, and nucleotide aminohydrolase activity on pyrimidine bases and their nucleosides and nucleotides. Only adenosine aminohydrolase activity was demonstrated. The enzyme was extracted from the cells and purified about 70-fold by liquid polymer two-phase fractionation, ammonium sulfate precipitation, and gel filtration. The enzyme has an apparent Km for adenosine of 4 × 10−5 M. The optimum pH is 7.5. The enzyme requires 4.0 M NaCl for maximal activity, but is irreversibly inactivated regardless of the concentration of NaCl when stored for extended periods of time.

Two types of xylanases of alkalophilic Bacillus sp. No. C-125

1985 ◽  
Vol 31 (6) ◽  
pp. 538-542 ◽  
Author(s):  
H. Honda ◽  
T. Kudo ◽  
Y. Ikura ◽  
K. Horikoshi
Keyword(s):  
Ammonium Sulfate ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Bacillus Sp ◽  
Ammonium Sulfate Precipitation ◽  
Molecular Weights ◽  
Protein Molecules ◽  
Activity Curve ◽  
Alkalophilic Bacillus ◽  
Hydrolysis Of

One alkalophilic Bacillus sp. strain C-125 (FERM No. 7344) was isolated from soil. From this organism, two types of xylanases, designated xylanase A and xylanase N, were purified by an ammonium sulfate precipitation followed by Biogel P-30 gel filtration, DEAE-cellulose chromatography, and Sephadex G-75 gel filtration. The molecular weights of xylanase A and N were estimated as 43 000 and 16 000, respectively. Immunological experiments indicated that xylanase A and xylanase N were entirely different protein molecules. Xylanase N was most active at pH 6.0–7.0, but xylanase A had a very broad pH activity curve (pH 6–10) and was still active even at pH 12.0. The maximum hydrolysis of xylan by the enzymes was about 25%. Both enzymes split xylan and yielded xylobiose and higher oligosaccharides but could hydrolyze neither xylobiose nor xylotriose. Trans xylosidation activities were detected in both enzymes.

scholarly journals Vitamin A and carotenoids. The enzymic conversion of β-carotene into retinal in hog intestinal mucosa

1969 ◽  
Vol 114 (4) ◽  
pp. 689-694 ◽  
Author(s):  
Noel H. Fidge ◽  
Frank Rees Smith ◽  
DeWitt S. Goodman
Keyword(s):  
Intestinal Mucosa ◽  
Gel Filtration ◽  
Maximal Activity ◽  
Enzyme Preparations ◽  
Optimum Ph ◽  
Tween 40 ◽  
Ph Range ◽  
Optimum Ph Range ◽  
Β Carotene

The conversion of β-carotene into retinal was studied in vitro with enzyme preparations from homogenates of hog intestinal mucosa. The hog mucosal enzyme was purified about 27-fold by precipitation with ammonium sulphate, chromatography on DEAE-Sephadex and gel filtration on Sephadex G-200. The reaction displayed a narrow optimum pH range (approx. 7·8–8·2). The enzyme was stimulated strongly by the addition of thiols, and was inhibited by thiol inhibitors and by the chelating agents αα′-bipyridyl and o-phenanthroline. The reaction required the addition of an appropriate detergent (or bile salt); maximal activity was obtained by addition of an appropriate combination of detergents and lipid (specifically Tween 40, sodium glycocholate and sphingomyelin). The reaction displayed Michaelis kinetics with Km1·3×10−6m and Vmax.1·1nmole of retinal formed/hr. (for 0·7mg. of enzyme protein). The properties of the hog enzyme are similar to those previously reported for a less purified rat enzyme preparation.

scholarly journals PURIFICATION AND SOME PROPERTIES OF ALPHA-AMYLASE OF PEAR FRUIT

HortScience ◽  
1991 ◽  
Vol 26 (6) ◽  
pp. 723E-723
Author(s):  
Shiao J. Li ◽  
Tim Facteau ◽  
Paul M. Chen
Keyword(s):  
Ammonium Sulfate ◽  
Time Course ◽  
Specific Activity ◽  
Alpha Amylase ◽  
Acetate Buffer ◽  
Pear Fruit ◽  
Ammonium Sulfate Precipitation ◽  
Freeze Dried ◽  
Optimum Ph ◽  
The Difference

Alpha-amylase was purified from freeze-dried pear fruit by extraction at pH 7.40 with Tris, Acetate and Imidazole buffer followed by differential ammonium sulfate precipitation and desalting column. The specific activity of the enzyme was increased 5.68 fold during purification. The optimum pH was 5.64 in Acetate buffer. The difference in the time course of alpha-amylase was observed between freeze-dried and fresh samples.

scholarly journals Production and Purification of the Endoglucanase Enzyme from Local Isolate Aspergillus fumigatus HBF356

2021 ◽  
Vol 12 (4) ◽  
pp. 4337-4347
Keyword(s):  
Aspergillus Fumigatus ◽  
Gel Filtration ◽  
Exchange Chromatography ◽  
Feed Additive ◽  
Optimum Temperature ◽  
Ammonium Sulfate Precipitation ◽  
Gentamicin Sulfate ◽  
Local Isolate ◽  
Optimum Ph ◽  
Very High

In this study, endoglucanase (EG) from local isolate Aspergillus fumigatus HBF356 was produced and purified using ammonium sulfate precipitation, gel filtration chromatography, and ion-exchange chromatography. The molecular weight of the pure EG was determined as 95 kDa. The optimum pH of the purified EG was determined as 4.0 and the optimum temperature as 60°C. It has been observed that the enzyme had a very high thermostability and preserved 75.8% of its activity after 240 hours of incubation at 50 °C. At the same time, the effect of veterinary drugs (gentamicin sulfate and enrofloxacin) on the activity of the EG was investigated. The activity of EG was inhibited by gentamicin sulfate while that was activated with enrofloxacin. The results of this study can give information about the potential of EG from Aspergillus fumigatus HBF356 using as a feed additive and its interaction with animal drugs.

A Bacteriocin Produced by Lactobacillus brevis KLDS1.0373 Isolated from “Jiaoke”, a Traditional Fermented Cream from China

2011 ◽  
Vol 183-185 ◽  
pp. 1132-1136
Author(s):  
Hui Wang ◽  
Han Sheng Gong ◽  
Xiang Chen Meng ◽  
Li Li Man
Keyword(s):  
Molecular Weight ◽  
Ammonium Sulfate ◽  
Inner Mongolia ◽  
Gel Filtration ◽  
Apparent Molecular Weight ◽  
Lactobacillus Brevis ◽  
Ammonium Sulfate Precipitation ◽  
Gel Filtration Chromatography

A bacteriocin produced by Lactobacillus brevis KLDS1.0373 which was isolated from “Jiaoke”, a traditional, naturally fermented cream from Inner Mongolia in China was reported in this article. The bacteriocin was partially purified by ammonium sulfate precipitation followed by sequential gel filtration chromatography, and the apparent molecular weight of the partially purified bacteriocin was estimated at approximately 3.8 kDa.

Production of fibrinolytic protease from a halobacterium Bacillus licheniformis VITLMS isolated from marine sponges of Rameshwaram coast, India

2020 ◽  
Vol 16 ◽  
Author(s):  
Lalith Kumar .CK ◽  
Merlyn Keziah.S ◽  
Hemalatha. M ◽  
Mohanapriya.A ◽  
Mohanasrinivasan.V ◽  
...  
Keyword(s):  
Ammonium Sulfate ◽  
Bacillus Licheniformis ◽  
Fibrinolytic Activity ◽  
Gel Filtration ◽  
Marine Sponges ◽  
Clot Lysis ◽  
Malt Extract ◽  
Ammonium Sulfate Precipitation ◽  
Fibrinolytic Enzymes ◽  
Lysis Activity

Background: Marine bacteria serve as excellent sources of therapeutic enzymes, metabolites and natural products, which possess novel therapeutic properties. Increasing death rates due to cardiovascular diseases urges for cost effective production of fibrinolytic enzyme. Methods: Marine sponge samples were screened for potent fibrinolytic producing bacteria. Primary screening was done for protease production, clot lysis activity. Secondary screening was done for casein plasminogen activity and fibrinolytic activity. The strain which had potent fibrinolytic activity among them was further subjected to morphological, biochemical and molecular characterization. Media optimization was carried to enhance enzyme production. The enzyme produced was subjected to purification using ammonium sulfate precipitation, gel filtration and characterized using HPLC and FTIR analysis Results: Sponge was identified to be Desmapsamma anchorata. Thirteen bacterial isolates were isolated from the sponge sample. . The 16S rRNA sequencing revealed that the potential strain had 99% similarity with Bacillus licheniformis. Amongst the isolates most were found to be morphologically identical to Bacillus genus. Gram’s staining and SEM analysis of the potent isolate was performed to identify the spore formation and rod shaped morphology of the bacteria. The optimal temperature and pH for production of the enzyme was 37 °C and 8 respectively. The carbon source maltose and nitrogen sources were malt extract and yeast extract were found to be optimal. The optimum incubation time was found to be both 4 to 5 days. The crude supernatant was purified with ammonium sulfate precipitation and gel filtration chromatography. The retention time of 11.3 min and presence of functional groups show the purity of the enzyme. The partially purified enzyme showed 96.4 % clot lysis in artificial clot lysis activity. Conclusion: Although the secretion of fibrinolytic enzymes from Bacillus species is not new, based on our investigation there are no reports regarding Bacillus licheniformis being isolated from marine sponges. However, there are reports of Bacillus licheniformis secreting fibrinolytic enzymes isolated from fermented food samples This study identifies marine environment as a potential source of new exploration for drug discovery.

scholarly journals Extraction and Partial Purification of Mouse Uterine Alkaline Phosphatase

1979 ◽  
Vol 32 (2) ◽  
pp. 153 ◽  
Author(s):  
RN Murdoch ◽  
DJ Kay ◽  
WJ Capper
Keyword(s):  
Alkaline Phosphatase ◽  
Ammonium Sulfate ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Partial Purification ◽  
Ammonium Sulfate Precipitation ◽  
Pregnant Mice ◽  
Triton X

Alkaline phosphatase in uterine homogenates from day 7 pregnant mice was solubilized using 0�2 % (v/v) Triton X-100 and extracted with 20% (v/v) n-butanol. The procedure, which resulted in 182- fold purification, included ammonium sulfate precipitation, DEAE-cellulose anion exchange chromatography and Sephadex 0200 gel filtration.

Extraction and Partial Purification of Protease from Bilimbi (Averrhoa bilimbi L.)

2013 ◽  
Vol 10 (2) ◽  
pp. 29
Author(s):  
Normah Ismail ◽  
Nur' Ain Mohamad Kharoe
Keyword(s):  
Molecular Weight ◽  
Protein Content ◽  
Ammonium Sulfate ◽  
Molecular Weight Distribution ◽  
Weight Distribution ◽  
Protease Activity ◽  
Protein Concentration ◽  
Temperature Stability ◽  
Partial Purification ◽  
Ammonium Sulfate Precipitation

Unripe and ripe bilimbi (Averrhoa bilimbi L.) were ground and the extracted juices were partially purified by ammonium sulfate precipitation at the concentrations of 40 and 60% (w/v). The collected proteases were analysed for pH, temperature stability, storage stability, molecular weight distribution, protein concentration and protein content. Protein content of bilimbi fruit was 0.89 g. Protease activity of both the unripe and ripe fruit were optimum at pH 4 and 40°C when the juice were purified at 40 and 60% ammonium sulfate precipitation. A decreased in protease activity was observed during the seven days of storage at 4°C. Molecular weight distribution indicated that the proteases protein bands fall between IO to 220 kDa. Protein bands were observed at 25, 50 and 160 kDa in both the unripe and ripe bilimbi proteases purified with 40% ammonium sulfate, however, the bands were more intense in those from unripe bilimbi. No protein bands were seen in proteases purified with 60% ammonium sulfate. Protein concentration was higher for proteases extracted with 40% ammonium sulfate at both ripening stages. Thus, purification using 40% ammonium sulfate precipitation could be a successful method to partially purify proteases from bilimbi especially from the unripe stage. 

scholarly journals A set of simple methods for detection and extraction of laminarinase

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ananthamurthy Koteshwara ◽  
Nancy V. Philip ◽  
Jesil Mathew Aranjani ◽  
Raghu Chandrashekhar Hariharapura ◽  
Subrahmanyam Volety Mallikarjuna
Keyword(s):  
Ammonium Sulfate ◽  
Ammonium Sulphate ◽  
Gold Standard ◽  
Direct Detection ◽  
Low Cost ◽  
Direct Analysis ◽  
Reducing Sugar ◽  
Ammonium Sulfate Precipitation ◽  
Simple Method ◽  
Functional Step

AbstractA carefully designed ammonium sulfate precipitation will simplify extraction of proteins and is considered to be a gold standard among various precipitation methods. Therefore, optimization of ammonium sulfate precipitation can be an important functional step in protein purification. The presence of high amounts of ammonium sulphate precludes direct detection of many enzymatically active proteins including reducing sugar assays (e.g. Nelson-Somogyi, Reissig and 3,5-dinitrosalicylic acid methods) for assessing carbohydrases (e.g. laminarinase (β (1–3)-glucanohydrolase), cellulases and chitinases). In this study, a simple method was developed using laminarin infused agarose plate for the direct analysis of the ammonium sulphate precipitates from Streptomyces rimosus AFM-1. The developed method is simple and convenient that can give accurate results even in presence of ammonium sulfate in the crude precipitates. Laminarin is a translucent substrate requiring the use of a stain to visualize the zones of hydrolysis in a plate assay. A very low-cost and locally available fluorescent optical fabric brightener Tinopal CBS-X has been used as a stain to detect the zones of hydrolysis. We also report simple methods to prepare colloidal chitin and cell free supernatant in this manuscript.

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