TRANSAMINATION IN PLANTS: THE SPECIFICITY OF AN AMINOTRANSFERASE FROM MUNG BEAN

1965 ◽  
Vol 43 (6) ◽  
pp. 723-730 ◽  
Author(s):  
Oluf L. Gamborg
Keyword(s):  
Amino Acids ◽  
Ammonium Sulfate ◽  
Mung Bean ◽  
Deae Cellulose ◽  
Phaseolus Aureus ◽  
Ammonium Sulfate Precipitation ◽  
Competitive Inhibitors

A study has been made of the specificity of an aminotransferase from mung bean (Phaseolus aureus Roxb.). The enzyme was purified 40- to 60-fold by using Sephadex G-50, ammonium sulfate precipitation, DEAE-cellulose, and hydroxylapatite. In the presence of pyruvate the enzyme transaminated a number of cyclic and aliphatic amino acids. Some of the better substrates were lysine, arginine, ornithine, glutamine, methionine, leucine, 4-fiuorophenyl-alanine, phenylalanine, tyrosine, tryptophan, 3,4-dihydroxyphenylalanine, and γ-phenylbutyrine. Threonine, serine, and glycine were not transaminated. Lysine, methionine, and glutamate were competitive inhibitors of the transamination of phenylalanine.

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.

A GENERAL METHOD FOR THE ISOLATION AND PURIFICATION OF PHOSVITIN FROM VERTEBRATE EGGS

1966 ◽  
Vol 44 (12) ◽  
pp. 1647-1655 ◽  
Author(s):  
R. A. Wallace ◽  
D. W. Jared ◽  
A. Z. Eisen
Keyword(s):  
Molecular Weight ◽  
Ammonium Sulfate ◽  
Deae Cellulose ◽  
The Other ◽  
Chemical Analyses ◽  
Ammonium Sulfate Precipitation ◽  
Isolation And Purification ◽  
Complex Ii ◽  
Vertebrate Eggs ◽  
General Method

A general method has been developed for the isolation and purification of phosvitin from vertebrate eggs. The method is detailed in three steps consisting of (i) isolation of a phosvitin–lipovitellin complex; (ii) ammonium sulfate precipitation of the lipovitellin; and (iii) DEAE-cellulose chromatography of the remaining phosvitin. Phosvitin was isolated from the eggs of five representative vertebrates (lamprey, trout, frog, turtle, and chicken), and chemical analyses together with sedimentation studies were performed on the samples. Preparations were obtained with the lowest N/P ratios reported to date. The analytical results also suggested that trout phosvitin has approximately half the molecular weight (24,000) of the other proteins and that "purified" phosvitin may still be heterogeneous.

Purification of Oidiodendron kalrai proteases

1976 ◽  
Vol 22 (3) ◽  
pp. 327-333 ◽  
Author(s):  
P. M. Cino ◽  
R. P. Tewari
Keyword(s):  
Ammonium Sulfate ◽  
Proteolytic Activity ◽  
Column Chromatography ◽  
Crude Extract ◽  
Proteolytic Enzymes ◽  
Deae Cellulose ◽  
Ammonium Sulfate Precipitation ◽  
Proteolytic Activities ◽  
Yeast Phase ◽  
Cellulose Column

Oidiodendron kalrai yeast-phase cells demonstrate proteolytic activity. Some of the proteolytic enzymes of the crude extract were purified by a combination of ammonium sulfate precipitation, Sephadex G-200, and diethylaminoethyl (DEAE) cellulose column chromatography. At least six proteins exhibiting a range of proteolytic activities could be identified by these procedures. Purity of the enzyme fractions obtained from the DEAE-cellulose columns was tested by running polyacrylamide gels.

The Carbohydrate Moiety of Mung Bean Vicilin

1976 ◽  
Vol 3 (6) ◽  
pp. 763 ◽  
Author(s):  
MC Ericson ◽  
MJ Chrispeels
Keyword(s):  
Mung Bean ◽  
Deae Cellulose ◽  
Indirect Evidence ◽  
Carbohydrate Moiety ◽  
Side Chain ◽  
Phaseolus Aureus ◽  
Molecular Weights ◽  
Molar Ratios ◽  
Peptide Moiety ◽  
Carbohydrate Unit

Mung bean (Phaseolus aureus Roxb.) vicilin, purified by DEAE-cellulose chromatography, is a glycoprotein (0.2% glucosamine and 1% mannose) which contains four different polypeptides with molecular weights of 63 500, 50000, 29 500 and 24 000 in the molar ratios 1 : 5 : 1 : 1. Two of these polypeptides (50 000 and 29 500) stain positively for carbohydrate. When vicilin is digested with pronase, the carbohydrate can be recovered as a glycopeptide containing mannose and glucosamine in the carbohydrate moiety and alanine, threonine and aspartic acid (or asparagine) in the peptide moiety. The total amount of carbohydrate (1.2 %) and the size of the carbohydrate unit (12 residues) suggest that each vicilin molecule contains only one carbohydrate side chain. Data indicate that this chain is attached via a glucosamine-asparagine link. The results raise the possibility that the presence of the two small polypeptides in vicilin preparations is the result of the breakdown of the major larger one. Indirect evidence suggests that vicilin may be a tetramer of four subunits, each with a molecular weight of 50 000, and that only one subunit has a carbohydrate side chain.

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.

scholarly journals SOME PROTEINS FROM THE MUNG BEAN, PHASEOLUS AUREUS ROXBURGH

1920 ◽  
Vol 44 (2) ◽  
pp. 303-317
Author(s):  
Carl O. Johns ◽  
Henry C. Waterman
Keyword(s):  
Mung Bean ◽  
Phaseolus Aureus

scholarly journals INVESTIGATIONS ON THE MUNG BEAN (PHASEOLUS AUREUS ROXBURGH)

1948 ◽  
Vol 175 (1) ◽  
pp. 377-383
Author(s):  
W. Edward Belton ◽  
Cecile A. Hoover
Keyword(s):  
Mung Bean ◽  
Phaseolus Aureus
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