Beta-galactosidase from a cold-adapted bacterium: purification, characterization and application for lactose hydrolysis.
(2002) In Applied Microbiology and Biotechnology 58(3). p.313-321- Abstract
- The enzyme beta-galactosidase was purified from a cold-adapted organism isolated from Antarctica. The organism was identified as a psychrotrophic Pseudoalteromonas sp. The enzyme was purified with high yields by a rapid purification scheme involving extraction in an aqueous two-phase system followed by hydrophobic interaction chromatography and ultrafiltration. The beta-galactosidase was optimally active at pH 9 and at 26 degrees C when assayed with o-nitrophenyl-beta-D-galactopyranoside as substrate for 2 min. The enzyme activity was highly sensitive to temperature above 30 degrees C and was undetectable at 40 degrees C. The cations Na+, K+, Mg2+ and Mn2+ activated the enzyme while Ca2+, Hg2+, Cu2+ and Zn2+ inhibited activity. The shelf... (More)
- The enzyme beta-galactosidase was purified from a cold-adapted organism isolated from Antarctica. The organism was identified as a psychrotrophic Pseudoalteromonas sp. The enzyme was purified with high yields by a rapid purification scheme involving extraction in an aqueous two-phase system followed by hydrophobic interaction chromatography and ultrafiltration. The beta-galactosidase was optimally active at pH 9 and at 26 degrees C when assayed with o-nitrophenyl-beta-D-galactopyranoside as substrate for 2 min. The enzyme activity was highly sensitive to temperature above 30 degrees C and was undetectable at 40 degrees C. The cations Na+, K+, Mg2+ and Mn2+ activated the enzyme while Ca2+, Hg2+, Cu2+ and Zn2+ inhibited activity. The shelf life of the pure enzyme at 4 degrees C was significantly enhanced in the presence of 0.1% (w/v) polyethyleneimine. The pure beta-galactosidase was also evaluated for lactose hydrolysis. More than 50% lactose hydrolysis was achieved in 8 h in buffer at an enzyme concentration of 1 U/ml, and was increased to 70% in the presence of 0.1% (w/v) polyethyleneimine. The extent of lactose hydrolysis was 40-50% in milk. The enzyme could be immobilized to Sepharose via different chemistries with 60-70% retention of activity. The immobilized enzyme was more stable and its ability to hydrolyze lactose was similar to that of the soluble enzyme. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/107427
- author
- Fernandes, S ; Geueke, B ; Delgado, Osvaldo LU ; Coleman, J and Hatti-Kaul, Rajni LU
- organization
- publishing date
- 2002
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Applied Microbiology and Biotechnology
- volume
- 58
- issue
- 3
- pages
- 313 - 321
- publisher
- Springer
- external identifiers
-
- wos:000174611500008
- pmid:11935182
- scopus:0036191816
- pmid:11935182
- ISSN
- 1432-0614
- DOI
- 10.1007/s00253-001-0905-4
- language
- English
- LU publication?
- yes
- id
- 7fea5e8f-6df0-4027-9649-ad0149267b7e (old id 107427)
- date added to LUP
- 2016-04-01 16:07:33
- date last changed
- 2022-01-28 17:28:11
@article{7fea5e8f-6df0-4027-9649-ad0149267b7e,
abstract = {{The enzyme beta-galactosidase was purified from a cold-adapted organism isolated from Antarctica. The organism was identified as a psychrotrophic Pseudoalteromonas sp. The enzyme was purified with high yields by a rapid purification scheme involving extraction in an aqueous two-phase system followed by hydrophobic interaction chromatography and ultrafiltration. The beta-galactosidase was optimally active at pH 9 and at 26 degrees C when assayed with o-nitrophenyl-beta-D-galactopyranoside as substrate for 2 min. The enzyme activity was highly sensitive to temperature above 30 degrees C and was undetectable at 40 degrees C. The cations Na+, K+, Mg2+ and Mn2+ activated the enzyme while Ca2+, Hg2+, Cu2+ and Zn2+ inhibited activity. The shelf life of the pure enzyme at 4 degrees C was significantly enhanced in the presence of 0.1% (w/v) polyethyleneimine. The pure beta-galactosidase was also evaluated for lactose hydrolysis. More than 50% lactose hydrolysis was achieved in 8 h in buffer at an enzyme concentration of 1 U/ml, and was increased to 70% in the presence of 0.1% (w/v) polyethyleneimine. The extent of lactose hydrolysis was 40-50% in milk. The enzyme could be immobilized to Sepharose via different chemistries with 60-70% retention of activity. The immobilized enzyme was more stable and its ability to hydrolyze lactose was similar to that of the soluble enzyme.}},
author = {{Fernandes, S and Geueke, B and Delgado, Osvaldo and Coleman, J and Hatti-Kaul, Rajni}},
issn = {{1432-0614}},
language = {{eng}},
number = {{3}},
pages = {{313--321}},
publisher = {{Springer}},
series = {{Applied Microbiology and Biotechnology}},
title = {{Beta-galactosidase from a cold-adapted bacterium: purification, characterization and application for lactose hydrolysis.}},
url = {{http://dx.doi.org/10.1007/s00253-001-0905-4}},
doi = {{10.1007/s00253-001-0905-4}},
volume = {{58}},
year = {{2002}},
}