TY - JOUR
T1 - Stereospecific production of 9R-hydroxy-10E,12Z-octadecadienoic acid from linoleic acid by recombinant Escherichia coli cells expressing 9R-lipoxygenase from Nostoc sp. SAG 25.82
AU - Kim, Kyoung Rok
AU - Seo, Min Ho
AU - Park, Jin Byung
AU - Oh, Deok Kun
N1 - Funding Information:
This study was supported by grants from the Bio-industry Technology Development Program, Ministry for Food, Agriculture, Forestry and Fisheries ( No. 112002-3 ) and the Korea Healthcare Technology R&D Project, Ministry for Health & Welfare, Republic of Korea ( No. 2012-009 ).
PY - 2014/6
Y1 - 2014/6
N2 - One of the most significant properties of lipoxygenase (LOX) as a biocatalyst is its stereo-selective oxygenation. In this study, the stereo-specific production of 9R-hydroxy-10E,12Z-octadecadienoic acid (9R-HODE) from linoleic acid was achieved using whole recombinant Escherichia coli cells expressing LOX from Nostoc sp. SAG 25.82. The optimal conditions for the production of 9R-HODE were pH 7.5, 25 °C, 40 g l-1 cells, 15 g l-1 linoleic acid, 2% (v/v) methanol, 1 working volume/oxygen volume/min (vvm) oxygenation rate, and 250 rpm agitation speed in 500 ml-baffled flask containing a working volume of 50 ml. Under these optimized conditions, whole recombinant cells expressing 9R-LOX protein produced 14.3 g l-1 9R-HODE for 1 h, with a conversion yield of 95% (w/w) and a productivity of 14.3 g l-1 h-1. The oxygen supply method significantly influenced stereo- and regio-selectivity of the oxygenation of linoleic acid. Among the oxygen supply methods tested, oxygenation (1 vvm) with agitation (250 rpm) resulted in the highest 9R/13S-HODE ratio of the products at 96:4. This is the first application using whole recombinant cells harboring R-specific LOX for the stereo-selective production of an R-specific hydroxy fatty acid.
AB - One of the most significant properties of lipoxygenase (LOX) as a biocatalyst is its stereo-selective oxygenation. In this study, the stereo-specific production of 9R-hydroxy-10E,12Z-octadecadienoic acid (9R-HODE) from linoleic acid was achieved using whole recombinant Escherichia coli cells expressing LOX from Nostoc sp. SAG 25.82. The optimal conditions for the production of 9R-HODE were pH 7.5, 25 °C, 40 g l-1 cells, 15 g l-1 linoleic acid, 2% (v/v) methanol, 1 working volume/oxygen volume/min (vvm) oxygenation rate, and 250 rpm agitation speed in 500 ml-baffled flask containing a working volume of 50 ml. Under these optimized conditions, whole recombinant cells expressing 9R-LOX protein produced 14.3 g l-1 9R-HODE for 1 h, with a conversion yield of 95% (w/w) and a productivity of 14.3 g l-1 h-1. The oxygen supply method significantly influenced stereo- and regio-selectivity of the oxygenation of linoleic acid. Among the oxygen supply methods tested, oxygenation (1 vvm) with agitation (250 rpm) resulted in the highest 9R/13S-HODE ratio of the products at 96:4. This is the first application using whole recombinant cells harboring R-specific LOX for the stereo-selective production of an R-specific hydroxy fatty acid.
KW - 9R-Hydroxy-10E,12Z-octadecadienoic acid
KW - Lipoxygenase
KW - Nostoc sp. SAG 25.82
KW - Oxygen supply
KW - Stereo-selective oxygenation
UR - http://www.scopus.com/inward/record.url?scp=84898076553&partnerID=8YFLogxK
U2 - 10.1016/j.molcatb.2014.03.009
DO - 10.1016/j.molcatb.2014.03.009
M3 - Article
AN - SCOPUS:84898076553
SN - 1381-1177
VL - 104
SP - 56
EP - 63
JO - Journal of Molecular Catalysis B: Enzymatic
JF - Journal of Molecular Catalysis B: Enzymatic
ER -