Purification and characterization of heat-stable exo-inulinase from Streptomyces sp.

Sharma, Arun Dev; Gill, Prabhjot Kaur

doi:10.1016/j.jfoodeng.2006.04.008

Cited by 44 publications

(29 citation statements)

References 25 publications

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“…(45 kDa) [16]. The enzyme had optimal activity at 55˚C and was stable at temperature up to 50 and still retained about 80% of its activity at 55˚C (Figure 5).…”

Section: Duction Of Inulinases From Various Microorganisms Including mentioning

confidence: 96%

“…The seed culture (10%, V/V) was inoculated into the basal medium (Czapek's Dox medium) contained (gl rpm at 30˚C for 3 days. After that the mycelium was removed by centrifugation at 8000 rpm for 10 min and the culture supernatant was assayed for inulinase activity as described by Sharma and Gill [16] in which the reaction was performed at pH 6.0 and 60˚C.…”

Section: Isolation Of Streptomyces and Screening For Inulinase-producmentioning

confidence: 99%

“…Inulinase activity was determined as described by Sharma and Gill [16] with minor modification. The reaction mixture containing 50 µl of enzyme sample, 55 µl of 50 mM sodium phosphate buffer pH 6.0 and 95 µl of 1% inulin suspended in 50 mM sodium phosphate buffer pH 6.0 was incubated at 55˚C for 20 min.…”

Section: Enzyme Assaymentioning

confidence: 99%

“…It is different from the inulinase from Streptomyces sp., the only purified and characterized inulinase from Streptomyces being reported so far, which is exoinulinase [16].…”

Section: Duction Of Inulinases From Various Microorganisms Including mentioning

confidence: 99%

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Production, Purification and Characterization of Inulinase from a Newly Isolated <i>Streptomyces</i> sp. CP01

Laowklom¹,

Chantanaphan²,

Pinphanichakarn³

2012

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Inulinase is an enzyme catalyzing the hydrolysis of inulin, a plant reserve polysaccharide, into fructoses and fructooligosaccharides which are widely used as food additives. Here we report inulinase from a newly isolated Streptomyces as in the past decade there have been very few reports on inulinases from Streptomyces, especially purification and characterization of these enzymes. Out of 371 Streptomyces isolates, Streptomyces sp. CP01 produced highest inulinase activity of 0.50 U/ml. The enzyme activity was increased to 1.60 U/ml when CP01 was cultivated under the optimal conditions which consisted of using basal medium (Czapek's Dox) containing 1% (w/v) inulin extract from Jerusalem artichoke's root tubers and 0.7% (w/v) tryptone at pH8, shaking at 200 rpm and 28˚C for 24 h. The enzyme was purified from culture filtrate to about 67-fold purity by (NH 4 ) 2 SO 4 precipitation followed by four consecutive column chromatography steps. The purified enzyme is a single peptide with approximate molecular mass of 73 kDa as analyzed by gel filtration and 70.8 kDa as assessed by SDS-PAGE. The enzyme is optimally active at 55˚C and pH 6.0, however it still possesses more than 80% of the maximal activity at pH ranging from 5.5 to 9.0. It is stable at temperature up to 50˚C and at broad range of pH from 5.0 to 9.0 for 30 min. Its K m and V max values for inulin were 2.34 mM and 440 μmol·min -1 ·mg -1 , respectively. This enzyme has potential for industrial application as it is active at moderately high temperature and wide range of pH.

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“…(45 kDa) [16]. The enzyme had optimal activity at 55˚C and was stable at temperature up to 50 and still retained about 80% of its activity at 55˚C (Figure 5).…”

Section: Duction Of Inulinases From Various Microorganisms Including mentioning

confidence: 96%

Section: Isolation Of Streptomyces and Screening For Inulinase-producmentioning

confidence: 99%

Section: Enzyme Assaymentioning

confidence: 99%

“…It is different from the inulinase from Streptomyces sp., the only purified and characterized inulinase from Streptomyces being reported so far, which is exoinulinase [16].…”

Section: Duction Of Inulinases From Various Microorganisms Including mentioning

confidence: 99%

See 2 more Smart Citations

Production, Purification and Characterization of Inulinase from a Newly Isolated <i>Streptomyces</i> sp. CP01

Laowklom¹,

Chantanaphan²,

Pinphanichakarn³

2012

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“…The fi nal enzyme preparation was homogeneous on SDS-PAGE, with a molecular mass of 55 kDa (Fig. 1) (13)(14)(15)(16)(17)(18). Five exoinulinases from Aspergillus fi cuum showed the same molecular mass of 74 kDa and three endoinulinases had a molecular mass of 64 kDa (19).…”

Section: Purifi Cation Of Inulinasementioning

confidence: 98%

Purification and Characterization of an Endoinulinase from Xanthomonas campestris pv. phaseoli KM 24 Mutant

Naidoo

Kumar

Sharma

et al. 2015

Food Technol. Biotechnol.

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SummaryAn extracellular endoinulinase from Xanthomonas campestris pv. phaseoli KM 24 mutant was purifi ed to homogeneity by gel fi ltration chromatography and showed a specifi c activity of 119 U/mg. The optimum pH and temperature of the purifi ed enzyme were found to be 6.0 and 50 °C, respectively. The enzyme was stable up to 60 °C, retaining 60 % of residual activity for 30 min, but inactivated rapidly above 60 °C. The enzyme was found to be stable at pH=6-9 when it retained 100 % of its residual activity. The Lineweaver-Burk plot showed that the apparent K m and v max values of the inulinase when using inulin as a substrate were 1.15 mg/mL and 0.15 μM/min, respectively, whereas the k cat value was found to be 0.145 min -1 . The calculated catalytic effi ciency of the enzyme was found to be 0.126 (mg·min)/mL. The purifi ed inulinase can be used in the production of high fructose syrups.

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Expression of exo‐inulinase gene from Aspergillus niger 12 in E. coli strain Rosetta‐gami B (DE3) and its characterization

Yedahalli

Rehmann

Bassi

2016

Biotechnology Progress

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Inulin is a linear carbohydrate polymer of fructose subunits (2-60) with terminal glucose units, produced as carbon storage in selected plants. It cannot directly be taken up by most microorganisms due to its large size, unless prior hydrolysis through inulinase enzymes occurs. The hydrolyzed inulin can be taken up by microbes and/or recovered and used industrially for the production of high fructose syrup, inulo-oligosaccharides, biofuel, and nutraceuticals. Cell-free enzymatic hydrolysis would be desirable for industrial applications, hence the recombinant expression, purification and characterization of an Aspergillus niger derived exo-inulinase was investigated in this study. The eukaroyototic exo-inulinase of Aspergillus niger 12 has been expressed, for the first time, in an E. coli strain [Rosetta-gami B (DE3)]. The molecular weight of recombinant exo-inulinase was estimated to be ∼81 kDa. The values of Km and Vmax of the recombinant exo-inulinase toward inulin were 5.3 ± 1.1 mM and 402.1 ± 53.1 µmol min(-1) mg(-1) protein, respectively. Towards sucrose the corresponding values were 12.20 ± 1.6 mM and 902.8 ± 40.2 µmol min(-1) mg(-1) protein towards sucrose. The S/I ratio was 2.24 ± 0.7, which is in the range of native inulinase. The optimum temperature and pH of the recombinant exo-inulinase towards inulin was 55°C and 5.0, while they were 50°C and 5.5 towards sucrose. The recombinant exo-inulinase activity towards inulin was enhanced by Cu(2+) and reduced by Fe(2+) , while its activity towards sucrose was enhanced by Co(2+) and reduced by Zn(2+) . © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:629-637, 2016.

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Purification and characterization of heat-stable exo-inulinase from Streptomyces sp.

Cited by 44 publications

References 25 publications

Production, Purification and Characterization of Inulinase from a Newly Isolated <i>Streptomyces</i> sp. CP01

Production, Purification and Characterization of Inulinase from a Newly Isolated <i>Streptomyces</i> sp. CP01

Purification and Characterization of an Endoinulinase from Xanthomonas campestris pv. phaseoli KM 24 Mutant

Expression of exo‐inulinase gene from Aspergillus niger 12 in E. coli strain Rosetta‐gami B (DE3) and its characterization

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Purification and characterization of heat-stable exo-inulinase from Streptomyces sp.

Cited by 44 publications

References 25 publications

Production, Purification and Characterization of Inulinase from a Newly Isolated &lt;i&gt;Streptomyces&lt;/i&gt; sp. CP01

Production, Purification and Characterization of Inulinase from a Newly Isolated &lt;i&gt;Streptomyces&lt;/i&gt; sp. CP01

Purification and Characterization of an Endoinulinase from Xanthomonas campestris pv. phaseoli KM 24 Mutant

Expression of exo‐inulinase gene from Aspergillus niger 12 in E. coli strain Rosetta‐gami B (DE3) and its characterization

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Product

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Production, Purification and Characterization of Inulinase from a Newly Isolated <i>Streptomyces</i> sp. CP01

Production, Purification and Characterization of Inulinase from a Newly Isolated <i>Streptomyces</i> sp. CP01