One-pot bioconversion of sucrose to trehalose using enzymatic sequential reactions in combined cross-linked enzyme aggregates.
Amylosucrase (AS), maltooligosyltrehalose synthase (MTS), and maltooligosyltrehalose trehalohydrolase (MTH) were used in combined cross-linked enzyme aggregates (combi-CLEAs) to achieve one-step bioconversion of sucrose to trehalose. Combi-CLEAs of three enzymes were successfully established with acetone and glutaraldehyde (GA) as a precipitant and a cross-linker, respectively. The optimum enzyme ratio was 8:0.5:0.5 (AS, 4mg:MTS, 0.25mg:MTH, 0.25mg). To improve trehalose production, bovine serum albumin was co-aggregated with enzymes as a proteic feeder. The trehalose production yield of combi-CLEAs was about 8% in each cycle on the basis of substrate added up to 400mM. Finally, the combi-CLEAs used in this experiment showed reusability of about five cycles without any activity loss.
Graduate School of Biotechnology, and Institute of Life Science and Resources, Kyung Hee University, Yongin 446-701, Republic of Korea.
Dong-Hyun
DH
Jung
Jong-Hyun
JH
Jung
Dong-Ho
DH
Seo
Suk-Jin
SJ
Ha
Dong-Keon
DK
Kweon
Cheon-Seok
CS
Park
D000096:Acetone / Q000737:chemistry; D000818:Animals; D002417:Cattle; D011232:Chemical Precipitation; D003432:Cross-Linking Reagents / Q000737:chemistry; D005959:Glucosidases / Q000737:chemistry / Q000378:metabolism*; D005964:Glucosyltransferases / Q000737:chemistry / Q000378:metabolism*; D005976:Glutaral / Q000737:chemistry; D012710:Serum Albumin, Bovine; D013395:Sucrose / Q000378:metabolism*; D014199:Trehalose / Q000378:metabolism*
D003432:Cross-Linking Reagents; D000096:Acetone; D012710:Serum Albumin, Bovine; D013395:Sucrose; D014199:Trehalose; D005964:Glucosyltransferases; C099084:maltooligosyl trehalose synthase; C019700:amylosucrase; D005959:Glucosidases; C099156:maltooligosyl trehalose trehalohydrolase; D005976:Glutaral