Advanced

A thermostable phytase from Bacillus sp. MD2: cloning, expression and high-level production in Escherichia coli.

Tran, Thi Thuy; Mamo, Gashaw LU ; Mattiasson, Bo LU and Hatti-Kaul, Rajni LU (2010) In Journal of Industrial Microbiology & Biotechnology1996-01-01+01:00 37(3). p.279-287
Abstract
Phytase is used as a feed additive for degradation of antinutritional phytate, and the enzyme is desired to be highly thermostable for it to withstand feed formulation conditions. A Bacillus sp. MD2 showing phytase activity was isolated, and the phytase encoding gene was cloned and expressed in Escherichia coli. The recombinant phytase exhibited high stability at temperatures up to 100 degrees C. A higher enzyme activity was obtained when the gene expression was done in the presence of calcium chloride. Production of the enzyme by batch- and fed-batch cultivation in a bioreactor was studied. In batch cultivation, maintaining dissolved oxygen at 20-30% saturation and depleting inorganic phosphate below 1 mM prior to induction by IPTG... (More)
Phytase is used as a feed additive for degradation of antinutritional phytate, and the enzyme is desired to be highly thermostable for it to withstand feed formulation conditions. A Bacillus sp. MD2 showing phytase activity was isolated, and the phytase encoding gene was cloned and expressed in Escherichia coli. The recombinant phytase exhibited high stability at temperatures up to 100 degrees C. A higher enzyme activity was obtained when the gene expression was done in the presence of calcium chloride. Production of the enzyme by batch- and fed-batch cultivation in a bioreactor was studied. In batch cultivation, maintaining dissolved oxygen at 20-30% saturation and depleting inorganic phosphate below 1 mM prior to induction by IPTG resulted in over 10 U/ml phytase activity. For fed-batch cultivation, glucose concentration was maintained at 2-3 g/l, and the phytase expression was increased to 327 U/ml. Induction using lactose during fed-batch cultivation showed a lag phase of 4 h prior to an increase in the phytase activity to 71 U/ml during the same period as IPTG-induced production. Up to 90% of the total amount of expressed phytase leaked out from the E. coli cells in both IPTG- and lactose-induced fed-batch cultivations. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Industrial Microbiology & Biotechnology1996-01-01+01:00
volume
37
issue
3
pages
279 - 287
publisher
Springer
external identifiers
  • wos:000274436100008
  • pmid:19997958
  • scopus:76849101050
ISSN
1367-5435
DOI
10.1007/s10295-009-0671-3
language
English
LU publication?
yes
id
9b2e38d2-583b-483d-987d-22306583f96d (old id 1523729)
date added to LUP
2010-01-15 10:22:27
date last changed
2018-05-29 09:31:12
@article{9b2e38d2-583b-483d-987d-22306583f96d,
  abstract     = {Phytase is used as a feed additive for degradation of antinutritional phytate, and the enzyme is desired to be highly thermostable for it to withstand feed formulation conditions. A Bacillus sp. MD2 showing phytase activity was isolated, and the phytase encoding gene was cloned and expressed in Escherichia coli. The recombinant phytase exhibited high stability at temperatures up to 100 degrees C. A higher enzyme activity was obtained when the gene expression was done in the presence of calcium chloride. Production of the enzyme by batch- and fed-batch cultivation in a bioreactor was studied. In batch cultivation, maintaining dissolved oxygen at 20-30% saturation and depleting inorganic phosphate below 1 mM prior to induction by IPTG resulted in over 10 U/ml phytase activity. For fed-batch cultivation, glucose concentration was maintained at 2-3 g/l, and the phytase expression was increased to 327 U/ml. Induction using lactose during fed-batch cultivation showed a lag phase of 4 h prior to an increase in the phytase activity to 71 U/ml during the same period as IPTG-induced production. Up to 90% of the total amount of expressed phytase leaked out from the E. coli cells in both IPTG- and lactose-induced fed-batch cultivations.},
  author       = {Tran, Thi Thuy and Mamo, Gashaw and Mattiasson, Bo and Hatti-Kaul, Rajni},
  issn         = {1367-5435},
  language     = {eng},
  number       = {3},
  pages        = {279--287},
  publisher    = {Springer},
  series       = {Journal of Industrial Microbiology & Biotechnology1996-01-01+01:00},
  title        = {A thermostable phytase from Bacillus sp. MD2: cloning, expression and high-level production in Escherichia coli.},
  url          = {http://dx.doi.org/10.1007/s10295-009-0671-3},
  volume       = {37},
  year         = {2010},
}