Cloning, Expression, and Purification of the His6-Tagged Thermostable β-Galactosidase from Pyrococcus woesei in Escherichia coli and Some Properties of the Isolated Enzyme

Daabrowski, S; Sobiewska, Gabriela; Maciuńska, Jadwiga; Synowiecki, J.; Kur, Józef

doi:10.1006/prep.2000.1231

Cited by 36 publications

(20 citation statements)

References 16 publications

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“…The reason for using this antibody was that the available anti-c-Myb antibodies were directed against the COOH terminus of the protein, which is deleted in our recombinant Myb-HIS, and because the histidine tag does not usually participate in the function of recombinant proteins (35)(36)(37)(38)(39)(40). Moreover, the Penta-HIS antibody easily detected the recombinant protein in Western blot analysis (data not shown).…”

Section: Mutagenesis Of Mbs-4 or Mutation Of C-myb Abrogates Mybdepenmentioning

confidence: 99%

Characterization of the c-Myb-responsive Region and Regulation of the Human Type I Collagen α2 Chain Gene by c-Myb

Luchetti

Paroncini

Majlingovà

et al. 2003

Journal of Biological Chemistry

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Section: Mutagenesis Of Mbs-4 or Mutation Of C-myb Abrogates Mybdepenmentioning

confidence: 99%

Characterization of the c-Myb-responsive Region and Regulation of the Human Type I Collagen α2 Chain Gene by c-Myb

Luchetti

Paroncini

Majlingovà

et al. 2003

Journal of Biological Chemistry

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“…[19À21] Although thermophilic bacteria can be good candidates for the development of thermostable b-galactosidase, most native thermostable enzymes give very low yield and are therefore difficult to purify. [22] A 1359 bp fragment was obtained from the genomic DNA of the bacterial consortium YTY-70 by using degenerate PCR primers, designed from the conserved regions of various thermostable b-galactosidase sequences. This fragment contained an open reading frame (ORF) encoding 452 amino acids (GenBank accession number JX531650).…”

Section: Cloning Of B-galactosidase Genes From Bacterial Consortium Ymentioning

confidence: 99%

Characterization of a thermostable recombinant β-galactosidase from a thermophilic anaerobic bacterial consortium YTY-70

Liu

Zhao

Deng

et al. 2015

Biotechnology & Biotechnological Equipment

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In the present study, a thermophilic anaerobic bacterial consortium YTY-70 that produced thermostable b-galactosidase was isolated from a hot spring in Yongtai (Fujian, China). Based on 16S rRNA sequences, we concluded that Caldicellulosiruptor was the predominant genus in the studied bacterial consortium. Subsequently, a thermostable b-galactosidase gene was cloned and expressed in Escherichia coli as a fusion protein with glutathione S-transferase. The recombinant enzyme exhibited a maximum activity at an optimum pH of 7.0 and an optimum temperature of 75 C. It had a high activity across a broad pH range (between pH 5.0 and pH 9.0) and high thermal stability at temperatures up to 90 C. The activity of the recombinant b-galactosidase was enhanced by TritonX-100, Tween-20 and 1 mmol/L 2-mercaptoethanol, while it was inhibited by ethylenediaminetetraacetic acid, sodium dodecyl sulphate, phenylmethylsulphonyl fluoride and 10 mmol/L 2-Me. The enzyme's catalytic function was enhanced by the following metal ions: Na C , K Fe 3C and Li C . The thermostable b-galactosidase might be a promising candidate for use in food industries, particularly for the dairy industry and biosynthesis of galacto-oligosaccharides, due to its high thermostability and broad pH range.

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“…Even though β-galactosidases from thermophilic (Dąabrowsky et al 2000;Ladero et al 2002) and psychrophilic organisms (Cavicchioli et al 2002;Nakagawa et al 2006) are attractive, the mesophilic enzymes from Aspergillus and Kluyveromyces, both having GRAS (generally recognized as safe) status, are the most adequate for industrial use (Illanes et al 1993). The former are thermostable and have a pH optimum between 3.5 and 5.5 , being well suited for the hydrolysis of acid whey and whey permeate; the latter, less stable and active at neutral pH, are preferred for the hydrolysis of sweet whey and milk (van GriethuysenDilber et al 1988).…”

Section: Upgrading By Enzymatic Hydrolysismentioning

confidence: 98%

Whey upgrading by enzyme biocatalysis

Illanes¹

2011

Electro Journal of Biotech

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Whey is a co-product of processes for the production of cheese and casein that retains most of the lactose content in milk. World production of whey is estimated around 200 million tons per year with an increase rate of about 2%/per year. Milk production is seasonal, so surplus whey is unavoidable. Traditionally, whey producers have considered it as a nuisance and strategies of whey handling have been mostly oriented to their more convenient disposal. This vision has been steadily evolving because of the upgrading potential of whey major components (lactose and whey proteins), but also because of more stringent regulations of waste disposal. Only the big cheese manufacturing companies are in the position of implementing technologies for their recovery and upgrading, so there is a major challenge in incorporating medium and small size producers to a platform of whey utilization, conciliating industrial interest with environmental protection within the framework of sustainable development. Within this context, among the many technological options for whey upgrading, transformation of whey components by enzyme biocatalysis appears as prominent. In fact, enzymes are green catalysts that can perform a myriad of transformation reactions under mild conditions and with strict specificity, so reducing production costs and environmental burden. This review pretends to highlight the impact of biocatalysis within a platform of whey upgrading. Technological options are shortly reviewed and then an in-depth and critical appraisal of enzyme technologies for whey upgrading is presented, with a special focus on newly developed enzymatic processes of organic synthesis, where the added value is high, being then a powerful driving force for industrial implementation.

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Cloning, Expression, and Purification of the His6-Tagged Thermostable β-Galactosidase from Pyrococcus woesei in Escherichia coli and Some Properties of the Isolated Enzyme

Cited by 36 publications

References 16 publications

Characterization of the c-Myb-responsive Region and Regulation of the Human Type I Collagen α2 Chain Gene by c-Myb

Characterization of the c-Myb-responsive Region and Regulation of the Human Type I Collagen α2 Chain Gene by c-Myb

Characterization of a thermostable recombinant β-galactosidase from a thermophilic anaerobic bacterial consortium YTY-70

Whey upgrading by enzyme biocatalysis

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