Biotechnology and Bioprocess Engineering 16: 1134-1143 (2011)
DOI 10.1007/s12257-011-0513-9
Isolation and Characterization of the New Klebsiella pneumoniae J2B
Strain Showing Improved Growth Characteristics with Reduced
Lipopolysaccharide Formation
Mariadhas Valan Arasu, Vinod Kumar, Somasundar Ashok, Hyohak Song, Chelladurai Rathnasingh, Hee Jong Lee,
Doyoung Seung, and Sunghoon Park
Received: 13 October 2011 / Accepted: 13 October 2011
© The Korean Society for Biotechnology and Bioengineering and Springer 2011
Abstract Klebsiella pneumoniae is a suitable biocatalyst
for the production of 1,3-propanediol (1,3-PDO) and 3-
hydroxypropionic acid (3-HP) from glycerol. However, its
commercial applications have been impeded due to its poor
growth characteristics and the excessive production of
lipopolysaccharide (LPS). To overcome these limitations, a
new K. pneumoniae J2B (KpJ2B) strain was isolated from
municipal waste anaerobic digester samples. The shake
flask cultivation of this new strain under aerobic conditions
showed a specific growth rate of 0.92/h, which is 1.13
times higher than that achieved using the well studied K.
pneumoniae DSMZ2026 (KpDSMZ). When the new strain
was grown in a bioreactor under aerobic conditions using
a fed-batch mode for 36 h, the biomass concentration (4.03
g/L CDW) and productivity (0.15 g/L/h) were almost 2.2
times higher than the corresponding values with KpDSMZ.
Growth was accompanied by the production of 1,3-PDO
(186 mM), lactic acid (235 mM), ethanol (170 mM), and
acetic acid (92.2 mM) at significant levels, indicating the
resistance of the strain to the inhibitory effects of these
metabolites. A comparison of the SEM images and 2-keto-
3-deoxyoctonate content (KpJ2B, 1.4 µg/g CDW; KpDSMZ,
1.9 µg/g CDW) confirmed the lower LPS content in the
KpJ2B strain. Furthermore, this new isolate exhibited
higher sensitivity towards a range of antibiotics and better
sedimentation properties than the KpDSMZ strain. This
suggests that KpJ2B is an attractive strain for industrial
applications.
Keywords: Klebsiella pneumoniae J2B, Klebsiella pneu-
moniae DSMZ2026, 1,3-propanediol, growth, lipopolysac-
charides
1. Introduction
Glycerol is an important chemical feedstock in the food
and cosmetic industries. In the past, its production from
plant oils and petroleum was limited and the price was
expensive as above US $1,000 per ton. Recently, due to the
rapid growth of biodiesel industry, its supply, especially
crude glycerol, increased drastically and, as a result, the
market prices have decreased substantially [1]. The crude
glycerol contains many impurities and cannot be used for
the food and cosmetic industries without extensive puri-
fication which is costly [2]. In comparison, its application
as a carbon substrate for microbial cultivation is not limited
by the presence of these impurities. Therefore, the
biological production of important chemicals, such as 1,3-
propanediol (1,3-PDO), 3-hydroxypropionic acid (3-HP)
and 1,3-dihydroxyacetone, from glycerol has attracted
considerable attention [3].
A number of microorganisms, such as Citrobacter fre-
undii , Klebsiella pnemoniae, Clostridium pasteurianum [4],
Clostridium butyricum, Enterobacter agglomerans [5], and
Enterobacter aerogenes [6], metabolize glycerol efficiently
to produce valuable chemical intermediates, such as 1,3-
PDO. Among these microbes, K. pneumoniae, has been
Mariadhas Valan Arasu, Vinod Kumar, Somasundar Ashok, Sunghoon Park
*
Department of Chemical and Biomolecular Engineering, Pusan National
University, Busan 609-735, Korea
Tel: +82-51-510-3386; Fax: +82-51-515-2716
E-mail: parksh@pusan.ac.kr
Hyohak Song, Chelladurai Rathnasingh, Hee Jong Lee, Doyoung Seung
Research and Development Center, GS Caltex Corporation, Daejeon 305-
380, Korea
RESEARCH PAPER