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Alkaliphilic Bacillus species and an alkaline active lipolytic enzyme

Vargas Calle, Virginia LU (2005)
Abstract
The Great Rift Valley running through Eastern Africa contains a large number of soda lakes. These unique lakes constitute among the most stable alkaline environments with a dense and varied population of microorganisms. Such microorganisms are invariable alkaliphiles and are considered to be a promising source of enzymes with unique features. Some enzymes produced by alkaliphilic microorganisms have found their way to industrial applications.Phylogenetic analysis and morphological and physiological characteristics of the isolates from a screening for lipase-producing microorganisms from a Kenyan alkaline soda lake showed some of them to be closely related to the Pseudomonas and Halomonas cluster sharing high similarity profile with... (More)
The Great Rift Valley running through Eastern Africa contains a large number of soda lakes. These unique lakes constitute among the most stable alkaline environments with a dense and varied population of microorganisms. Such microorganisms are invariable alkaliphiles and are considered to be a promising source of enzymes with unique features. Some enzymes produced by alkaliphilic microorganisms have found their way to industrial applications.Phylogenetic analysis and morphological and physiological characteristics of the isolates from a screening for lipase-producing microorganisms from a Kenyan alkaline soda lake showed some of them to be closely related to the Pseudomonas and Halomonas cluster sharing high similarity profile with Halomonas desiderata. The remaining were Gram-positive, closely related to the Bacillus cluster, and were grouped with Bacillus halodurans, Bacillus alcalophilus and Bacillus licheniformis, respectively. A new alkaliphilic Bacillus sp. was identified during the screening and based on detailed studies we proposed a novel alkaliphilic, halotolerant species of the genus Bacillus with the name Bacillus bogoriensis.Among the alkaliphilic Bacillus species isolated, Bacillus halodurans was selected due to its remarkable enzymes studied so far. A lipolytic enzyme was cloned and overexpressed in Escherichia coli. Production of the enzyme was mainly in the form of inclusion bodies from which recovery of the active enzyme was achieved. Since E. coli proved not to be a suitable host for production of soluble enzyme, Pichia pastoris was tested as an alternative host and the production of the heterologous enzyme in a soluble form was also studied. The enzyme was characterized as a phospholipase, also displaying hydrolysis of soluble and insoluble fatty substrates, and esterification of fatty acids. (Less)
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author
supervisor
opponent
  • Dr Stougaard, Peter, Royal Veterinary and Agricultural University, Denmark
organization
publishing date
type
Thesis
publication status
published
subject
keywords
Biotechnology, soda lakes, alkaliphilic Bacillus sp., lipolytic enzyme, Bacillus bogoriensis, Bacillus halodurans, Bioteknik
pages
113 pages
publisher
Biotechnology, Lund University
defense location
Center for Chemistry and Chemical Engineering, Lecture hall C, Lund Institute of Technology
defense date
2005-10-10 10:30:00
external identifiers
  • other:ISRN: LUTKDH/TKBT-05/1087-SE
ISBN
91-89627-34-2
language
English
LU publication?
yes
id
a0620b40-7695-4696-99af-0d2ed0416de9 (old id 24875)
date added to LUP
2016-04-04 11:35:43
date last changed
2018-11-21 21:05:53
@phdthesis{a0620b40-7695-4696-99af-0d2ed0416de9,
  abstract     = {{The Great Rift Valley running through Eastern Africa contains a large number of soda lakes. These unique lakes constitute among the most stable alkaline environments with a dense and varied population of microorganisms. Such microorganisms are invariable alkaliphiles and are considered to be a promising source of enzymes with unique features. Some enzymes produced by alkaliphilic microorganisms have found their way to industrial applications.Phylogenetic analysis and morphological and physiological characteristics of the isolates from a screening for lipase-producing microorganisms from a Kenyan alkaline soda lake showed some of them to be closely related to the Pseudomonas and Halomonas cluster sharing high similarity profile with Halomonas desiderata. The remaining were Gram-positive, closely related to the Bacillus cluster, and were grouped with Bacillus halodurans, Bacillus alcalophilus and Bacillus licheniformis, respectively. A new alkaliphilic Bacillus sp. was identified during the screening and based on detailed studies we proposed a novel alkaliphilic, halotolerant species of the genus Bacillus with the name Bacillus bogoriensis.Among the alkaliphilic Bacillus species isolated, Bacillus halodurans was selected due to its remarkable enzymes studied so far. A lipolytic enzyme was cloned and overexpressed in Escherichia coli. Production of the enzyme was mainly in the form of inclusion bodies from which recovery of the active enzyme was achieved. Since E. coli proved not to be a suitable host for production of soluble enzyme, Pichia pastoris was tested as an alternative host and the production of the heterologous enzyme in a soluble form was also studied. The enzyme was characterized as a phospholipase, also displaying hydrolysis of soluble and insoluble fatty substrates, and esterification of fatty acids.}},
  author       = {{Vargas Calle, Virginia}},
  isbn         = {{91-89627-34-2}},
  keywords     = {{Biotechnology; soda lakes; alkaliphilic Bacillus sp.; lipolytic enzyme; Bacillus bogoriensis; Bacillus halodurans; Bioteknik}},
  language     = {{eng}},
  publisher    = {{Biotechnology, Lund University}},
  school       = {{Lund University}},
  title        = {{Alkaliphilic Bacillus species and an alkaline active lipolytic enzyme}},
  year         = {{2005}},
}