Bioconversion of D-galactose to D-tagatose: continuous packed bed reaction with an immobilized thermostable L-arabinose isomerase and efficient purification by selective microbial degradation.
The continuous enzymatic conversion of D-galactose to D-tagatose with an immobilized thermostable L-arabinose isomerase in packed-bed reactor and a novel method for D-tagatose purification were studied. L-arabinose isomerase from Thermoanaerobacter mathranii (TMAI) was recombinantly overexpressed and immobilized in calcium alginate. The effects of pH and temperature on D-tagatose production reaction catalyzed by free and immobilized TMAI were investigated. The optimal condition for free enzyme was pH 8.0, 60°C, 5 mM MnCl(2). However, that for immobilized enzyme was pH 7.5, 75°C, 5 mM MnCl(2). In addition, the catalytic activity of immobilized enzyme at high temperature and low pH was significantly improved compared with free enzyme. The optimum reaction yield with immobilized TMAI increased by four percentage points to 43.9% compared with that of free TMAI. The highest productivity of 10 g/L h was achieved with the yield of 23.3%. Continuous production was performed at 70°C; after 168 h, the reaction yield was still above 30%. The resultant syrup was then incubated with Saccharomyces cerevisiae L1 cells. The selective degradation of D-galactose was achieved, obtaining D-tagatose with the purity above 95%. The established production and separation methods further potentiate the industrial production of D-tagatose via bioconversion and biopurification processes.
State Key Laboratory of Microbial Technology, Shandong University, Jinan, Shandong, China.
Min
M
Liang
Min
M
Chen
Xinying
X
Liu
Yafei
Y
Zhai
Xian-wei
XW
Liu
Houcheng
H
Zhang
Min
M
Xiao
Peng
P
Wang
D019747:Aldose-Ketose Isomerases / Q000737:chemistry / Q000378:metabolism*; D001711:Biotransformation; D002712:Chlorides / Q000378:metabolism; D020536:Enzyme Activators / Q000378:metabolism; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005690:Galactose / Q000378:metabolism*; D006601:Hexoses / Q000302:isolation & purification* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D017895:Manganese Compounds / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D013696:Temperature; D046968:Thermoanaerobacter / Q000201:enzymology
D002712:Chlorides; D020536:Enzyme Activators; D004800:Enzymes, Immobilized; D006601:Hexoses; D017895:Manganese Compounds; D011994:Recombinant Proteins; D019747:Aldose-Ketose Isomerases; C019715:L-arabinose isomerase; C025340:manganese chloride; C030192:tagatose; D005690:Galactose