Directed evolution of a dextransucrase for increased constitutive activity and the synthesis of a highly branched dextran

被引:13
作者
Kang, HK
Seo, ES
Robyt, JF
Kim, D [1 ]
机构
[1] Chonnam Natl Univ, Inst Engn Res, Kwangju 500757, South Korea
[2] Chonnam Natl Univ, Fac Appl Chem Engn, Kwangju 500757, South Korea
[3] Chonnam Natl Univ, Inst Catalysis, Kwangju 500757, South Korea
[4] Iowa State Univ, Lab Carbohydrate Chem & Enzymol, Ames, IA 50011 USA
关键词
dextransucrase; directed bacterial evolution; constitutivity; Leuconostoc mesenteroides mutants; dextran; E. coli dextransucrase transformants;
D O I
10.1016/j.molcatb.2003.05.006
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
An Escherichia coli transformant (pDSRB742CK) was obtained from the DSRB742 clone by using ultrasoft X-rays for the expression of a dextransucrase. The enzyme differed in several aspects from DSRB742 dextransucrase: it (1) was constitutive; (2) was extracellular; (3) had 2.6 times greater activity (0.035 IU/ml and 0.23 IU/mg); and (4) synthesized a highly (15.6%) alpha-(1 --> 3) branched dextran. Seven nucleotides of the parent gene (dsrB742) were changed in the nucleotide sequence; four nucleotides were changed in the open reading frame (ORF) that resulted in a 30 amino acid deletion in the N-terminus. (C) 2003 Elsevier B.V. All rights reserved.
引用
收藏
页码:167 / 176
页数:10
相关论文
共 45 条
[1]   Sequence analysis of the gene encoding alternansucrase, a sucrose glucosyltransferase from Leuconostoc mesenteroides NRRL B-1355 [J].
Argüello-Morales, MA ;
Remaud-Simeon, M ;
Pizzut, S ;
Sarçabal, P ;
Willemot, RM ;
Monsan, P .
FEMS MICROBIOLOGY LETTERS, 2000, 182 (01) :81-85
[2]   SEQUENCE-ANALYSIS OF THE GENE FOR THE GLUCAN-BINDING PROTEIN OF STREPTOCOCCUS-MUTANS INGBRITT [J].
BANAS, JA ;
RUSSELL, RRB ;
FERRETTI, JJ .
INFECTION AND IMMUNITY, 1990, 58 (03) :667-673
[3]  
Blight Mark A., 1994, Current Opinion in Biotechnology, V5, P468, DOI 10.1016/0958-1669(94)90059-0
[4]   Induction and rejoining of DNA double-strand breaks in Chinese hamster V79-4 cells irradiated with characteristic aluminum K and copper L ultrasoft X rays [J].
Botchway, SW ;
Stevens, DL ;
Hill, MA ;
Jenner, TJ ;
ONeill, P .
RADIATION RESEARCH, 1997, 148 (04) :317-324
[5]   Molecular characterization of DSR-E, an α-1,2 linkage-synthesizing dextransucrase with two catalytic domains [J].
Bozonnet, S ;
Dols-Laffargue, M ;
Fabre, E ;
Pizzut, S ;
Remaud-Simeon, M ;
Monsan, P ;
Willemot, RM .
JOURNAL OF BACTERIOLOGY, 2002, 184 (20) :5753-5761
[6]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[7]   Expressing genes in different Escherichia coli compartments [J].
Cornelis, P .
CURRENT OPINION IN BIOTECHNOLOGY, 2000, 11 (05) :450-454
[8]   DEGRADATION AND FERMENTATION OF ALPHA-GLUCO-OLIGOSACCHARIDES BY BACTERIAL STRAINS FROM HUMAN COLON - IN-VITRO AND IN-VIVO STUDIES IN GNOTOBIOTIC-RATS [J].
DJOUZI, Z ;
ANDRIEUX, C ;
PELENC, V ;
SOMARRIBA, S ;
POPOT, F ;
PAUL, F ;
MONSAN, P ;
SZYLIT, O .
JOURNAL OF APPLIED BACTERIOLOGY, 1995, 79 (02) :117-127
[9]   NUCLEOTIDE-SEQUENCE OF A GLUCOSYLTRANSFERASE GENE FROM STREPTOCOCCUS-SOBRINUS MFE28 [J].
FERRETTI, JJ ;
GILPIN, ML ;
RUSSELL, RRB .
JOURNAL OF BACTERIOLOGY, 1987, 169 (09) :4271-4278
[10]   Molecular analyses of glucosyltransferase genes among strains of Streptococcus mutans [J].
Fujiwara, T ;
Terao, Y ;
Hoshino, T ;
Kawabata, S ;
Ooshima, T ;
Sobue, S ;
Kimura, S ;
Hamada, S .
FEMS MICROBIOLOGY LETTERS, 1998, 161 (02) :331-336