Optimal conditions for levan formation by an overexpressed recombinant levansucrase

被引:36
作者
Belghith, H [1 ]
Song, KB [1 ]
Kim, CH [1 ]
Rhee, SK [1 ]
机构
[1] KIST,KOREA RES INST BIOSCI & BIOTECHNOL,APPL MICROBIOL RES DIV,TAEJON 305600,SOUTH KOREA
关键词
D O I
10.1007/BF00143472
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
A genetically modified levansucrase, which contained His-affinity tag in its C-terminal, was constructed by PCR reaction using two synthetic primers. This modified protein was produced up to 30 % in total cell protein of E. coli, and was purified by a one-step affinity chromatography. The optimum pH for levan production was pH 5 and the optimum temperature was 0 degrees C. The higher velocity of levan formation within shorter enzyme reaction times was achieved by increasing the levels of enzyme concentration. The optimal sucrose concentration for levan production was around 20 %. Under these conditions, more than 50 g levan/l was produced.
引用
收藏
页码:467 / 472
页数:6
相关论文
共 15 条
[1]   LEVANSUCRASE OF BACILLUS-SUBTILIS - CHARACTERIZATION OF A STABILIZED FRUCTOSYL-ENZYME COMPLEX AND IDENTIFICATION OF AN ASPARTYL RESIDUE AS BINDING-SITE OF FRUCTOSYL GROUP [J].
CHAMBERT, R ;
GONZYTREBOUL, G .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1976, 71 (02) :493-508
[2]   PRODUCTION AND CHARACTERIZATION OF MICROBIAL LEVAN [J].
HAN, YW ;
CLARKE, MA .
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 1990, 38 (02) :393-396
[3]   PURIFICATION AND CHARACTERIZATION OF AN EXTRACELLULAR LEVANSUCRASE FROM PSEUDOMONAS-SYRINGAE PV PHASEOLICOLA [J].
HETTWER, U ;
GROSS, M ;
RUDOLPH, K .
JOURNAL OF BACTERIOLOGY, 1995, 177 (10) :2834-2839
[4]   CONTINUOUS CULTURE SYSTEM FOR PRODUCTION OF BIOPOLYMER LEVAN USING ERWINIA-HERBICOLA [J].
KEITH, J ;
WILEY, B ;
BALL, D ;
ARCIDIACONO, S ;
ZORFASS, D ;
MAYER, J ;
KAPLAN, D .
BIOTECHNOLOGY AND BIOENGINEERING, 1991, 38 (05) :557-560
[5]  
Maniatis T., 1982, MOL CLONING
[6]   IDENTIFICATION OF SACCHAROLYTIC ENZYMES OF ZYMOMONAS-MOBILIS CP4 [J].
OMULLAN, P ;
SZAKACSDOBOZI, M ;
EVELEIGH, DE .
BIOTECHNOLOGY LETTERS, 1991, 13 (02) :137-142
[7]  
REISS M, 1989, Chemie Mikrobiologie Technologie der Lebensmittel, V12, P1
[8]  
SONG KB, 1994, BIOTECHNOL LETT, V16, P1305
[9]  
SONG KB, 1993, BIOCHIM BIOPHYS ACTA, V1171, P320
[10]  
STUDIER FW, 1990, METHOD ENZYMOL, V185, P60