Cloning, Purification and Characterization of an NAD-Dependent d-Arabitol Dehydrogenase from Acetic Acid Bacterium, Acetobacter suboxydans

被引：17
作者：
Cheng, Hairong [1 ,2 ]
Li, Zilong [3 ]
Jiang, Ning [3 ]
Deng, Zixin [1 ,2 ]
机构：
[1] Shanghai Jiao Tong Univ, Lab Microbial Metab, Shanghai 200194, Peoples R China
[2] Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai 200194, Peoples R China
[3] Chinese Acad Sci, Ctr Microbial Biotechnol, Inst Microbiol, Beijing 100080, Peoples R China
来源：
PROTEIN JOURNAL | 2009年 / 28卷 / 06期
关键词：
Acetobacter suboxydans; D-Arabitol dehydrogenase; NAD(P); Oxidation/reduction; Polyol; GLUCONOBACTER-OXYDANS; ALCOHOL-DEHYDROGENASE; FUNCTIONAL EXPRESSION; POLYOL DEHYDROGENASE; ESCHERICHIA-COLI; CANDIDA-ALBICANS; GENE; XYLITOL; CRYSTALLIZATION; CONSTRUCTION;
D O I：
10.1007/s10930-009-9191-2
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：
d-Xylulose-forming d-arabitol dehydrogenase (aArDH) is a key enzyme in the bio-conversion of d-arabitol to xylitol. In this study, we cloned the NAD-dependent d-xylulose-forming d-arabitol dehydrogenase gene from an acetic acid bacterium, Acetobacter suboxydans sp. The enzyme was purified from A. suboxydans sp. and was heterogeneously expressed in Escherichia coli. The native or recombinant enzyme was preferred NAD(H) to NADP(H) as coenzyme. The active recombinant aArDH expressed in E. coli is a homodimer, whereas the native aArDH in A. suboxydans is a homotetramer. On SDS-PAGE, the recombinant and native aArDH give one protein band at the position corresponding to 28 kDa. The optimum pH of polyol oxidation and ketone reduction is found to be pH 8.5 and 5.5 respectively. The highest reaction rate is observed when d-arabitol is used as the substrate (K (m) = 4.5 mM) and the product is determined to be d-xylulose by HPLC analysis.
引用
收藏
页码：263 / 272
页数：10
相关论文
共 33 条
[1]   Membrane-bound sugar alcohol dehydrogenase in acetic acid bacteria catalyzes L-ribulose formation and NAD-dependent ribitol dehydrogenase is independent of the oxidative fermentation [J].
Adachi, O ;
Fujii, Y ;
Ano, Y ;
Moonmangmee, D ;
Toyama, H ;
Shinagawa, E ;
Theeragool, G ;
Lotong, N ;
Matsushita, K .
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 2001, 65 (01) :115-125
[2]   Crystallization and properties of NADPH-dependent L-sorbose reductase from Gluconobacter melanogenus IFO 3294 [J].
Adachi, O ;
Ano, Y ;
Moonmangmee, D ;
Shinagawa, E ;
Toyama, H ;
Theeragool, G ;
Lotong, N ;
Matsushita, K .
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 1999, 63 (12) :2137-2143
[3]   PURIFICATION AND PROPERTIES OF A POLYOL DEHYDROGENASE FROM CEPHALOSPORIUM-CHRYSOGENUS [J].
BIRKEN, S ;
PISANO, MA .
JOURNAL OF BACTERIOLOGY, 1976, 125 (01) :225-232
[4]   Identification of the operon for the sorbitol (glucitol) phosphoenolpyruvate:sugar phosphotransferase system in Streptococcus mutans [J].
Boyd, DA ;
Thevenot, T ;
Gumbmann, M ;
Honeyman, AL ;
Hamilton, IR .
INFECTION AND IMMUNITY, 2000, 68 (02) :925-930
[5]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[6]   His-tag impact on structure [J].
Carson, Mike ;
Johnson, David H. ;
McDonald, Heather ;
Brouillette, Christie ;
DeLucas, Lawrence J. .
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 2007, 63 :295-301
[7]  
Chang M, 2005, LASER FOCUS WORLD, V41, P67
[8]   Extremely rapid extraction of DNA from bacteria and yeasts [J].
Cheng, HR ;
Jiang, N .
BIOTECHNOLOGY LETTERS, 2006, 28 (01) :55-59
[9]   Molecular cloning and functional expression of D-arabitol dehydrogenase gene from Gluconobacter oxydans in Escherichia coli [J].
Cheng, HR ;
Jiang, N ;
Shen, A ;
Feng, YJ .
FEMS MICROBIOLOGY LETTERS, 2005, 252 (01) :35-42
[10]   Purification and characterization of two NAD-dependent alcohol dehydrogenases (ADHs) induced in the quinoprotein ADH-deficient mutant of Acetobacter pasteurianus SKU1108 [J].
Chinnawirotpisan, P ;
Matsushita, K ;
Toyama, H ;
Adachi, O ;
Limtong, S ;
Theeragool, G .
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 2003, 67 (05) :958-965
1234