Purification and characterization of cyclodextrin glucanotransferase from alkalophilic Bacillus sp. G1

Sian, Ho Kek; Said, Mamot; Hassan, Osman; Kamaruddin, Kamarulzaman; Ismail, Akram; Rahman, Roshanida A.; Mahmood, Nik Hasnaa Nik; Illias, Rosli Md.

doi:10.1016/j.procbio.2004.03.018

Cited by 74 publications

(57 citation statements)

References 32 publications

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“…CGTase activity was determined using the phenolphthalein assay 13 . One milliliter of substrate buffer containing 40 mg/ml soluble starch in 0.1 M phosphate buffer, pH 6.0 was added to 0.1 ml protein sample and incubated at 60 °C for 10 min.…”

Section: Methodsmentioning

confidence: 99%

Effect of the cross linkers’ concentration on the immobilization of recombinant Escherichia coli cells on hollow fiber membrane for excretion of cyclodextrin glucanotransferase (CGTase)

CheMan

Illias

Sulaiman

et al. 2017

Chem. Eng. Res. Bull.

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Abstract:The excretion of recombinant proteins into the culture medium is a preferred approach over cytoplasmic expression because of the high specific activity and ease of purification. However, cell lysis is one of a major problem during the excretion of recombinant protein due to the pressure build up in the periplasmic space through overproduction of the expressed recombinant protein. Cell immobilization is a promising solution for the improvement of protein excretion with reduction in cell lysis. In this study, the effect of cross linkers concentrations on the process of cell immobilization for the cyclodextringlucanotransferase (CGTase) excretion and cell viability were investigated. The hollow fiber membrane was treated using different concentrations (0.3, 0.67 and 1%) of cross linkers namely glutaraldehyde (GA), poly-L-lysine (PLL) and polyethylenimine (PEI). The optimum concentration of cross linkers was found to be 0.3% with the high excretion of CGTase. Interestingly, the immobilized cell on the non-treated hollow fiber membrane showed up to 15% increment of CGTase excretion with 55% reduction of cell lysis, as compared to the immobilized cells on the treated hollow fiber membrane. It could be concluded that, the low concentration of cross linkers exhibited the highest CGTase excretion. Moreover, the non-treated hollow fiber membrane is a promising approach for attachment of cells to the membrane without requiring treatment with any chemicals for the production of recombinant enzyme.

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Section: Methodsmentioning

confidence: 99%

Effect of the cross linkers’ concentration on the immobilization of recombinant Escherichia coli cells on hollow fiber membrane for excretion of cyclodextrin glucanotransferase (CGTase)

CheMan

Illias

Sulaiman

et al. 2017

Chem. Eng. Res. Bull.

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show abstract

“…G1 (Cao et al 2005) and B. stearothermophilus ET1 (Sian et al 2005). However, the mutants showed lower activity at pH 4.0 and 8.0, suggested that the mutants and wild type of b-CGTase required a near-neutral pH range to perform its reaction.…”

Section: Activity-temperature Profilementioning

confidence: 99%

“…Extreme pH values were not suitable for the enzyme to carry out cyclization activity. Most of the reported CGTases exhibited optimum pH range from 5.0 to 8.0 (Bovetto et al 1992;Chung et al 1998;Tachibana et al 1999;Sian et al 2005), but the enzyme from Brevi-bacterium sp. no.…”

Section: Activity-temperature Profilementioning

confidence: 99%

Improved activity of β-cyclodextrin glycosyltransferase from Bacillus sp. N-227 via mutagenesis of the conserved residues

Wang

Zhou

et al. 2017

3 Biotech

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b-Cyclodextrin glycosyltransferase (b-CGTase) belongs to the a-amylase family of enzymes and converts starch to cyclic oligosaccharides called b-cyclodextrins (b-CD). The b-CGTase from alkalophilic Bacillus sp. N-227 was separately mutagenized to give three site-directed bCGTase mutants, Y127F, R254F and D355R, that showed enhanced cyclization activity towards a starch substrate from 1.64 to 2.1-folds. Kinetic studies indicate that the mutants had higher affinity towards the substrate than the wild type b-CGTase. The Y127F mutant had the highest affinity which was indicated by the lowest K m of 15.30 mM and the highest catalytic activity. Increasing hydrophobicity around the catalytic center appeared to favor the cyclization activity of the mutants. The bCGTase and the three mutants showed optimal enzyme activity at 60°C and pH 6.0. All the enzymes were stable for at least 60 min across a wide pH range (5.0-7.0).

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“…As CGTase produces preferentially α-, β-and/or γ-CD they are called as α-, β-and/or γ-CGTase. A CGTase that produces only one type of CD is not known, but a CGTase which produces a high proportion of one type of cyclodextrin is industrially favorable because it reduces the cost of the purifying process [2] [6] [7].…”

Section: Introductionmentioning

confidence: 99%

Bovine Bone Charcoal as Support Material for Immobilization of <em>Bacillus firmus</em> Strain 37 and Production of Cyclomaltodextrin Glucanotransferase by Batch Fermentation in a Fluidized Bed

Silva¹,

Bieli²,

Valarini³

et al. 2018

ACES

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The process described in the present work uses air supplementation in a fluidized bed reactor containing Bacillus firmus strain 37 immobilized on active bovine bone charcoal, to produce by batch fermentation the enzyme CGTase (cyclomaltodextrin-glucanotransferase). Three different aeration rates were evaluated. The maximum CGTase activity was achieved after 120 hours of fermentation with aeration rate of 2 vvm and was equal to 2.48 U/mL. When 0.5 and 1 vvm were used the enzymatic activities achieved 1.1 and 0.57 U/mL, respectively. Bovine bone charcoal was characterized in terms of surface area, pore size and volume. To the best of our knowledge, the immobilization of microorganism cells in bovine bone charcoal for CGTase production has not been reported in the literature. Our results showed that fluidized bed reactor allows retaining high concentration of biomass, improving biomass-substrate contact and operation at low residence times, which resulted in improved enzyme production. Therefore, the process as proposed has great potential for industrial development.

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Purification and characterization of cyclodextrin glucanotransferase from alkalophilic Bacillus sp. G1

Cited by 74 publications

References 32 publications

Effect of the cross linkers’ concentration on the immobilization of recombinant Escherichia coli cells on hollow fiber membrane for excretion of cyclodextrin glucanotransferase (CGTase)

Effect of the cross linkers’ concentration on the immobilization of recombinant Escherichia coli cells on hollow fiber membrane for excretion of cyclodextrin glucanotransferase (CGTase)

Improved activity of β-cyclodextrin glycosyltransferase from Bacillus sp. N-227 via mutagenesis of the conserved residues

Bovine Bone Charcoal as Support Material for Immobilization of <em>Bacillus firmus</em> Strain 37 and Production of Cyclomaltodextrin Glucanotransferase by Batch Fermentation in a Fluidized Bed

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Purification and characterization of cyclodextrin glucanotransferase from alkalophilic Bacillus sp. G1

Cited by 74 publications

References 32 publications

Effect of the cross linkers’ concentration on the immobilization of recombinant Escherichia coli cells on hollow fiber membrane for excretion of cyclodextrin glucanotransferase (CGTase)

Effect of the cross linkers’ concentration on the immobilization of recombinant Escherichia coli cells on hollow fiber membrane for excretion of cyclodextrin glucanotransferase (CGTase)

Improved activity of β-cyclodextrin glycosyltransferase from Bacillus sp. N-227 via mutagenesis of the conserved residues

Bovine Bone Charcoal as Support Material for Immobilization of &lt;em&gt;Bacillus firmus&lt;/em&gt; Strain 37 and Production of Cyclomaltodextrin Glucanotransferase by Batch Fermentation in a Fluidized Bed

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Bovine Bone Charcoal as Support Material for Immobilization of <em>Bacillus firmus</em> Strain 37 and Production of Cyclomaltodextrin Glucanotransferase by Batch Fermentation in a Fluidized Bed