Permeabilization of Escherichia coli cells in the biotransformation of trimethylammonium compounds into L-carnitine

被引:48
作者
Cánovas, M [1 ]
Torroglosa, T [1 ]
Iborra, JL [1 ]
机构
[1] Univ Murcia, Dept Biochem & Mol Biol B & Immunol, Fac Chem, E-30100 Murcia, Spain
关键词
cell permeabilization; Escherichia coli; biotransformation; L-carnitine; D-carnitine; crotonobetaine;
D O I
10.1016/j.enzmictec.2004.07.023
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The activity of permeabilized cells of Escherichia coli strains during the biotransformation of trimethylamonium compounds, such as crotonobetaine and D-carnitine, into L-carnitine was determined. Permeabilization of cells did not affect the biotransformation of D-carnitine but improved the yield of L-carnitine obtained when crotonobetaine was used as biotransformation substrate (by nearly two-fold). A higher degree of cell activity with respect to control (45% yield) was observed with organic solvents such as acetone, ethanol, isopropyl alcohol and toluene (65-70% yield), detergents such as Triton X-100 and Tween 20 (70-75% yield) and polyethylenimine (80-89% yield). However, maximum cell activity was attained with 2% of the detergents, Triton X-100 and Tween 20, and 0.1% of polyethylenimine, using a temperature of 37 degrees C and 10-30min of treatment time, in which case a yield of 75-89% (100% increase) was obtained with E. coli 044 K74 and 85-94% (80% increase) with E. coli K38 T7-5KE32. Both detergents and polyethylenimine produced alterations in the outer membrane of treated cells since thin-sectioned bacteria were observed by transmision electron microscopy. NO L-carnitine dehydratase or carnitine racemase enzyme activity was detected within the extracellular medium when detergents and polyethyleneimine were used. Furthermore, transport studies showed that trimethyl ammonium compound carriers were not affected by permeabilization, while the rate of L-carnitime transport increased by 20%. (c) 2005 Elsevier Inc. All rights reserved.
引用
收藏
页码:300 / 308
页数:9
相关论文
共 27 条
[1]   Lactic acid permeabilizes gram-negative bacteria by disrupting the outer membrane [J].
Alakomi, HL ;
Skyttä, E ;
Saarela, M ;
Mattila-Sandholm, T ;
Latva-Kala, K ;
Helander, IM .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2000, 66 (05) :2001-2005
[2]   Effect of permeabilizers on antibiotic sensitivity of Pseudomonas aeruginosa [J].
Ayres, HM ;
Furr, JR ;
Russell, AD .
LETTERS IN APPLIED MICROBIOLOGY, 1999, 28 (01) :13-16
[3]   Biotransformation of D(+)-carnitine into L(-)-carnitine by resting cells of Escherichia coli O44 K74 [J].
Castellar, MR ;
Cánovas, M ;
Kleber, HP ;
Iborra, JL .
JOURNAL OF APPLIED MICROBIOLOGY, 1998, 85 (05) :883-890
[4]  
EICHLER K, 1996, J BACTERIOL, V13, P775
[5]   PERMEABILIZED CELLS [J].
FELIX, H .
ANALYTICAL BIOCHEMISTRY, 1982, 120 (02) :211-234
[6]   PERMEABILIZATION OF YEAST-CELLS (KLUYVEROMYCES) WITH ORGANIC-SOLVENTS [J].
FLORES, MV ;
VOGET, CE ;
ERTOLA, RJJ .
ENZYME AND MICROBIAL TECHNOLOGY, 1994, 16 (04) :340-346
[7]   Permeabilization of Yarrowia lipolytica cells by Triton X-100 [J].
Galabova, D ;
Tuleva, B ;
Spasova, D .
ENZYME AND MICROBIAL TECHNOLOGY, 1996, 18 (01) :18-22
[8]   Detergents improve xanthan yield and polymer quality in cultures of Xanthomonas campestris [J].
Galindo, E ;
Salcedo, G .
ENZYME AND MICROBIAL TECHNOLOGY, 1996, 19 (02) :145-149
[9]   Fluorometric assessment of Gram-negative bacterial permeabilization [J].
Helander, IM ;
Mattila-Sandholm, T .
JOURNAL OF APPLIED MICROBIOLOGY, 2000, 88 (02) :213-219
[10]   Permeabilizing action of polyethyleneimine on Salmonella typhimurium involves disruption of the outer membrane and interactions with lipopolysaccharide [J].
Helander, IM ;
Latva-Kala, K ;
Lounatmaa, K .
MICROBIOLOGY-UK, 1998, 144 :385-390