Purification and characterization of 5′-phosphodiesterase from germinated barley

Dhule, Santosh S.; Shetty, Premalata R.; Iyer, Jyoti L.

doi:10.1016/j.procbio.2006.03.041

Cited by 5 publications

(15 citation statements)

References 20 publications

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“…Fraction profiles of barley malt sprouts 5'-PDE and PME residual activities after acetone partial purification of crude enzyme preparation purified 5'-PDE was close to the K m (0.28 mM) of highly purified 5'-PDE from barley rootlets[8]. Dhule et al[13] found that K m values of purified 5'-PDE and PME from germinated barley were 0.33 mM and 0.23 mM, respectively. Compared with the K m values of the 5'-PDE from other plant sources, this K m is relatively high.…”

mentioning

confidence: 93%

Comparative Biochemical Characterization of 5′-Phosphodiesterase and Phosphomonoesterase from Barley Malt Sprouts

Beluhan

Marić

2011

Natural Product Communications

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A comparative biochemical characterization is described of two competing enzymes in the production of flavoring 5'-ribonucleotides, barley malt sprouts 5'-phosphodiesterase (5'-PDE) and phosphomonoesterase (PME). Fractionation of these two enzymes and partial purification of 5'-PDE were achieved by a combination of thermal treatments and precipitation with acetone. With synthetic substrates, under standard assay conditions, 5'-PDE and PME had maximum activities at pH 8.9, 70ºC and 55ºC, and K m of 0.26 mM and 0.19 mM, respectively. In the presence of 10 mM Mg 2+ ions, barley malt sprouts 5'-PDE was activated by up to 160% of the original activity, while PME was inhibited. Zn 2+ activated PME by up to 125 % of the original activity. Both enzymes were moderately inhibited after addition of Cu 2+ , Co 2+ , Ca 2+ , and Mn 2+ ions (10 mM), but, significantly, by addition of the chelating agent EDTA. In the absence of substrate and up to 80ºC, barley malt sprouts 5'-PDE showed excellent stability and retained 70% of its original activity at 70ºC after 120 min.

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mentioning

confidence: 93%

Comparative Biochemical Characterization of 5′-Phosphodiesterase and Phosphomonoesterase from Barley Malt Sprouts

Beluhan

Marić

2011

Natural Product Communications

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“…However, in general, the temperature optimum for 5'-PDE was found to be 70°C and below. This can be seen in 5'-PDE from germinated barley (Dhule et al, 2006), Penicillium citrinum (Ying et al, 2006), Aspergillus fumigatus (Luo et al, 2017) and malt rootlet (Zou et al, 2008) which had temperature optimums of 60°C, 65°C, 60°C and 70°C, respectively.…”

Section: Temperature Optimum and Stabilitymentioning

confidence: 99%

“…The homogenate was stirred for 30 minutes before filtering through a piece of muslin cloth. The filtrate was subjected to centrifugation at 10°C for 15 min at 12,5000 × g (Sartorius Model 3-18 k centrifuge, Sartorius AG, Weender Land Strasse, Gottingen, Germany; Dhule et al, 2006).…”

Section: Extraction Of Enzymes From Germinated Adzuki Beansmentioning

confidence: 99%

“…Previous studies have been done on germinating beans as sources of 5'-PDE such as barley, mung bean (Khutle et al, 2011), yellow soybean and black soybean (Utami et al, 2011). Also, 5'-PDE with different pH and temperature optima, as well as other characteristics, were reported for microorganisms such as Xanthomonas axonopodis (Lassila and Herschlag, 2008), Rhizobium legumniosarum (Jones et al, 2008), Aspergillus niger (Chohnan et al, 1994), Aspergillus fumigatus (Luo et al, 2017) and Escherichia coli (Miller et al, 2007); animals such as the cow (Rosemeyer et al, 2002) and plants as well, such as thale cress (Speath et al, 2007) and rose periwinkle flower (Aoyagi et al, 2006).…”

Section: Introductionmentioning

confidence: 99%

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Characterization of crude 5’-phosphodiesterase from germinated adzuki (Vigna angularis L.) beans

Pui

Mohammed²,

Ghazali

2020

Acta Sci Pol Technol Aliment

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Background. 5'-Phosphodiesterase (5'-PDE) is an enzyme that hydrolyzes RNA to form 5'-inosine monophosphate (5'-IMP) and 5'-guanosine monophosphate (5'-GMP). These 5'-nucleotides can function as flavor enhancers. Adzuki beans (Vigna angularis L.) are found to be high in 5'-PDE. Materials and methods. 5'-phosphodiesterase (5'-PDE) enzyme was characterized from adzuki beans, in which the optimum pH and temperature were determined. In addition, the stability of 5'-PDE was assessed at different pH and temperature. The effects of cations and EDTA were evaluated to characterize the 5'-PDE enzymes further. Results. The alkaline 5'-phosphodiesterase has an optimum pH of 8.5. This enzyme is also thermostable, with an optimum temperature of 80°C. The stability in terms of temperature and pH was also determined, and was found to be stable in the pH range of 7.0-8.5. This enzyme was found to retain more than 80% of its activity for 4 days at 60 and 65°C. In addition, the effects of 14 different metal ions, 4 types of detergents and ethylenediaminetetraacetic acid (EDTA) on 5'-PDE were studied.

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“…The large solid particles and starch were separated by centrifugation at 5000 rpm at 8-10 C for 15 min. To quantify the enzyme extraction, enzymatic assay was made to determine Alpha amylase activities 12,13 .…”

Section: Germination Of Mat Bean and Extraction Of Enzymementioning

confidence: 99%

Extraction and Characterization of Alpha Amylase from Phaseolus Aconitifolius.

Nerkar¹,

Chaudhari²,

Khutle.³

2011

JCPR

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The Alpha amylase breaks down long chain carbohydrates and that's why it is very useful in food and pharmaceutical industries. The aim of the present work is to extract and characterize the Alpha amylase from germinated Mat beans, for optimum pH and temperature for activity and stability, effect of metal ions, optimum substrate concentration, Km and V max. Mat beans were germinated and then extracted with Acetate buffer (pH 5.0). The amylase assay was based on the reduction in blue color intensity resulting from enzyme hydrolysis of starch. For optimization of pH for stability, buffer solutions from pH range of 3.5 to 10.0 were prepared. For determination of optimum temperature, the range used was 35 o C to 65 o C. Substrate solutions in range of 0.1% to 4.0% were prepared for optimum substrate concentration. Mat beans germinated for 3 days show maximum enzymatic activity (65.79U/ml). 95% of the original activity was retained in the pH range 7.0 to 8.0. The maximum activity of an enzyme was found to be at 55 C. 10mM Ca 2+ stimulated the enzyme activity up to 123%. Optimum substrate concentration was found to be 1.5%. Km was found to be 2.0 mM, and Vmax 100.0 nm/min. Alpha amylase extracted from germinated Mat beans follows Michaelis-Menten equation. It can be concluded that Mat bean is a good source of Alpha amylase and this enzyme could find promising application of hydrolysis of starch.

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Purification and characterization of 5′-phosphodiesterase from germinated barley

Cited by 5 publications

References 20 publications

Comparative Biochemical Characterization of 5′-Phosphodiesterase and Phosphomonoesterase from Barley Malt Sprouts

Comparative Biochemical Characterization of 5′-Phosphodiesterase and Phosphomonoesterase from Barley Malt Sprouts

Characterization of crude 5’-phosphodiesterase from germinated adzuki (Vigna angularis L.) beans

Extraction and Characterization of Alpha Amylase from Phaseolus Aconitifolius.

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