A new cold-adapted β-D-galactosidase from the Antarctic Arthrobacter sp. 32c – gene cloning, overexpression, purification and properties
Abstract Background The development of a new cold-active β-D-galactosidases and microorganisms that efficiently ferment lactose is of high biotechnological interest, particularly for lactose removal in milk and dairy products at low temperatures and for cheese whey bioremediation processes with simu...
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Online Access: | http://dx.doi.org/10.1186/1471-2180-9-151 https://link.springer.com/content/pdf/10.1186/1471-2180-9-151.pdf |
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crspringernat:10.1186/1471-2180-9-151 2023-05-15T14:13:10+02:00 A new cold-adapted β-D-galactosidase from the Antarctic Arthrobacter sp. 32c – gene cloning, overexpression, purification and properties Hildebrandt, Piotr Wanarska, Marta Kur, Józef 2009 http://dx.doi.org/10.1186/1471-2180-9-151 https://link.springer.com/content/pdf/10.1186/1471-2180-9-151.pdf en eng Springer Science and Business Media LLC BMC Microbiology volume 9, issue 1 ISSN 1471-2180 Microbiology (medical) Microbiology journal-article 2009 crspringernat https://doi.org/10.1186/1471-2180-9-151 2022-01-04T08:12:15Z Abstract Background The development of a new cold-active β-D-galactosidases and microorganisms that efficiently ferment lactose is of high biotechnological interest, particularly for lactose removal in milk and dairy products at low temperatures and for cheese whey bioremediation processes with simultaneous bio-ethanol production. Results In this article, we present a new β-D-galactosidase as a candidate to be applied in the above mentioned biotechnological processes. The gene encoding this β-D-galactosidase has been isolated from the genomic DNA library of Antarctic bacterium Arthrobacter sp. 32c, sequenced, cloned, expressed in Escherichia coli and Pichia pastoris , purified and characterized. 27 mg of β-D-galactosidase was purified from 1 L of culture with the use of an intracellular E. coli expression system. The protein was also produced extracellularly by P. pastoris in high amounts giving approximately 137 mg and 97 mg of purified enzyme from 1 L of P. pastoris culture for the AOX1 and a constitutive system, respectively. The enzyme was purified to electrophoretic homogeneity by using either one step- or a fast two step- procedure including protein precipitation and affinity chromatography. The enzyme was found to be active as a homotrimeric protein consisting of 695 amino acid residues in each monomer. Although, the maximum activity of the enzyme was determined at pH 6.5 and 50°C, 60% of the maximum activity of the enzyme was determined at 25°C and 15% of the maximum activity was detected at 0°C. Conclusion The properties of Arthrobacter sp. 32cβ-D-galactosidase suggest that this enzyme could be useful for low-cost, industrial conversion of lactose into galactose and glucose in milk products and could be an interesting alternative for the production of ethanol from lactose-based feedstock. Article in Journal/Newspaper Antarc* Antarctic Springer Nature (via Crossref) Antarctic The Antarctic BMC Microbiology 9 1 151 |
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Springer Nature (via Crossref) |
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English |
topic |
Microbiology (medical) Microbiology |
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Microbiology (medical) Microbiology Hildebrandt, Piotr Wanarska, Marta Kur, Józef A new cold-adapted β-D-galactosidase from the Antarctic Arthrobacter sp. 32c – gene cloning, overexpression, purification and properties |
topic_facet |
Microbiology (medical) Microbiology |
description |
Abstract Background The development of a new cold-active β-D-galactosidases and microorganisms that efficiently ferment lactose is of high biotechnological interest, particularly for lactose removal in milk and dairy products at low temperatures and for cheese whey bioremediation processes with simultaneous bio-ethanol production. Results In this article, we present a new β-D-galactosidase as a candidate to be applied in the above mentioned biotechnological processes. The gene encoding this β-D-galactosidase has been isolated from the genomic DNA library of Antarctic bacterium Arthrobacter sp. 32c, sequenced, cloned, expressed in Escherichia coli and Pichia pastoris , purified and characterized. 27 mg of β-D-galactosidase was purified from 1 L of culture with the use of an intracellular E. coli expression system. The protein was also produced extracellularly by P. pastoris in high amounts giving approximately 137 mg and 97 mg of purified enzyme from 1 L of P. pastoris culture for the AOX1 and a constitutive system, respectively. The enzyme was purified to electrophoretic homogeneity by using either one step- or a fast two step- procedure including protein precipitation and affinity chromatography. The enzyme was found to be active as a homotrimeric protein consisting of 695 amino acid residues in each monomer. Although, the maximum activity of the enzyme was determined at pH 6.5 and 50°C, 60% of the maximum activity of the enzyme was determined at 25°C and 15% of the maximum activity was detected at 0°C. Conclusion The properties of Arthrobacter sp. 32cβ-D-galactosidase suggest that this enzyme could be useful for low-cost, industrial conversion of lactose into galactose and glucose in milk products and could be an interesting alternative for the production of ethanol from lactose-based feedstock. |
format |
Article in Journal/Newspaper |
author |
Hildebrandt, Piotr Wanarska, Marta Kur, Józef |
author_facet |
Hildebrandt, Piotr Wanarska, Marta Kur, Józef |
author_sort |
Hildebrandt, Piotr |
title |
A new cold-adapted β-D-galactosidase from the Antarctic Arthrobacter sp. 32c – gene cloning, overexpression, purification and properties |
title_short |
A new cold-adapted β-D-galactosidase from the Antarctic Arthrobacter sp. 32c – gene cloning, overexpression, purification and properties |
title_full |
A new cold-adapted β-D-galactosidase from the Antarctic Arthrobacter sp. 32c – gene cloning, overexpression, purification and properties |
title_fullStr |
A new cold-adapted β-D-galactosidase from the Antarctic Arthrobacter sp. 32c – gene cloning, overexpression, purification and properties |
title_full_unstemmed |
A new cold-adapted β-D-galactosidase from the Antarctic Arthrobacter sp. 32c – gene cloning, overexpression, purification and properties |
title_sort |
new cold-adapted β-d-galactosidase from the antarctic arthrobacter sp. 32c – gene cloning, overexpression, purification and properties |
publisher |
Springer Science and Business Media LLC |
publishDate |
2009 |
url |
http://dx.doi.org/10.1186/1471-2180-9-151 https://link.springer.com/content/pdf/10.1186/1471-2180-9-151.pdf |
geographic |
Antarctic The Antarctic |
geographic_facet |
Antarctic The Antarctic |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_source |
BMC Microbiology volume 9, issue 1 ISSN 1471-2180 |
op_doi |
https://doi.org/10.1186/1471-2180-9-151 |
container_title |
BMC Microbiology |
container_volume |
9 |
container_issue |
1 |
container_start_page |
151 |
_version_ |
1766285583064236032 |