A secretion signal is present in the Collybia velutipes oxalate decarboxylase gene

被引:17
作者
Azam, M
Kesarwani, M
Natarajan, K
Datta, A
机构
[1] Natl Ctr Plant Genome Res, New Delhi 110067, India
[2] Jawaharlal Nehru Univ, Sch Life Sci, New Delhi 110067, India
关键词
oxalate decarboxylase; secretion signal; expression vector; Schizosaccharomyces pombe;
D O I
10.1006/bbrc.2001.6049
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
The oxalate decarboxylase (OXDC) gene from Collybia velutipes is overexpressed as an active form in Schizosaccharomyces pombe. The recombinant enzyme shows similar pH optima and stability, while substrate kinetic analysis shows a ninefold decrease in K-m value with respect to native OXDC. Most of the expressed protein was present in periplasm and remained firmly bound to cell-wall materials. However, 20% of enzyme expressed was secreted out into the medium suggesting the presence of a secretion signal (C. velutipes) in the oxalate decarboxylase gene. This secretion signal is associated with the N-terminal of OXDC as is evident by secretion of nonsecretory genes AmA1 and beta-galactosidase. An expression vector using this signal is constructed for expression and secretion of heterologous proteins in S. pombe. (C) 2001 Elsevier Science.
引用
收藏
页码:807 / 812
页数:6
相关论文
共 26 条
[1]  
ARNOLD WN, 1972, J BIOL CHEM, V247, P1161
[2]   PURIFICATION AND CHARACTERIZATION OF OXALYL-COENZYME-A DECARBOXYLASE FROM OXALOBACTER-FORMIGENES [J].
BAETZ, AL ;
ALLISON, MJ .
JOURNAL OF BACTERIOLOGY, 1989, 171 (05) :2605-2608
[3]   OXALATE, FORMATE, FORMAMIDE, AND METHANOL METABOLISM IN THIOBACILLUS-NOVELLUS [J].
CHANDRA, TS ;
SHETHNA, YI .
JOURNAL OF BACTERIOLOGY, 1977, 131 (02) :389-398
[4]   MULTIFUNCTIONAL YEAST HIGH-COPY-NUMBER SHUTTLE VECTORS [J].
CHRISTIANSON, TW ;
SIKORSKI, RS ;
DANTE, M ;
SHERO, JH ;
HIETER, P .
GENE, 1992, 110 (01) :119-122
[5]   TRANSFORMATION OF INTACT YEAST-CELLS TREATED WITH ALKALI CATIONS [J].
ITO, H ;
FUKUDA, Y ;
MURATA, K ;
KIMURA, A .
JOURNAL OF BACTERIOLOGY, 1983, 153 (01) :163-168
[6]  
JAMES LF, 1972, CLIN TOXICOL, V5, P243
[7]  
JONES EW, 1991, METHOD ENZYMOL, V194, P428
[8]   Oxalate decarboxylase from Collybia velutipes -: Molecular cloning and its overexpression to confer resistance to fungal infection in transgenic tobacco and tomato [J].
Kesarwani, M ;
Azam, M ;
Natarajan, K ;
Mehta, A ;
Datta, A .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2000, 275 (10) :7230-7238
[9]   EXTERNAL ENZYMES OF YEAST - THEIR NATURE AND FORMATION [J].
LAMPEN, JO .
ANTONIE VAN LEEUWENHOEK JOURNAL OF MICROBIOLOGY, 1968, 34 (01) :1-&
[10]  
Lamport D. T. A., 1980, The biochemistry of plants. A comprehensive treatise. Volume 3. Carbohydrates: structure and function., P501