Biotransformation of rutin to isoquercitrin using recombinant α-L-rhamnosidase from Bifidobacterium breve.
OBJECTIVE
To biotransform rutin into isoquercitrin.


RESULTS
A α-L-rhamnosidase from Bifidobacterium breve was produced by using Escherichia coli BL21 for biotransformation of rutin to isoquercitrin. The enzyme was purified by Ni(2+)-NTA chromatography to yield a soluble protein with a specific activity of 56 U protein mg(-1). The maximum enzyme activities were at pH 6.5, 55 °C, 20 mM rutin, and 1.2 U enzyme ml(-1). Under optimal conditions, the half-life of the enzyme was 96 h. The K m and V max values were 2.2 mM, 56.4 μmol mg(-1) min(-1) and 2.1 mM, 57.5 μmol mg(-1) min(-1) using pNP-Rha and rutin as substrates, respectively. The kinetic behavior indicated that the recombinant α-L-rhamnosidase has good catalytic performance for producing isoquercitrin. 20 mM rutin was biotransformed into 18.25 and 19.87 mM isoquercitrin after 60 and 240 min.


CONCLUSIONS
The specific biotransformation of rutin to isoquercitrin using recombinant α-L-rhamnosidase from B. breve is a feasible method for use in industrial processes.
College of Chemistry and Chemical Engineering, Hunan Institute of Engineering, Xiangtan, 411104, China, zhangru2002@hotmail.com.
Ru
R
Zhang
Bian-Ling
BL
Zhang
Tao
T
Xie
Gu-Cai
GC
Li
Yi
Y
Tuo
Yu-Ting
YT
Xiang
D001644:Bifidobacterium / Q000201:enzymology* / Q000235:genetics; D001711:Biotransformation; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D011794:Quercetin / Q000031:analogs & derivatives* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012431:Rutin / Q000378:metabolism*; D013696:Temperature
D011994:Recombinant Proteins; C016527:isoquercitrin; D012431:Rutin; D011794:Quercetin; D006026:Glycoside Hydrolases; C022525:alpha-L-rhamnosidase