One-pot bioconversion of sucrose to trehalose using enzymatic sequential reactions in combined cross-linked enzyme aggregates

• Published in: Bioresource technology Vol. 130; pp. 801 - 804
• Main Authors: Jung, Dong-Hyun, Jung, Jong-Hyun, Seo, Dong-Ho, Ha, Suk-Jin, Kweon, Dong-Keon, Park, Cheon-Seok
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier Ltd 01-02-2013; Elsevier
• Subjects: Acetone - chemistry; Amylosucrase; Animals; Bioconversions. Hemisynthesis; Biological and medical sciences; Biotechnology; Cattle; Chemical Precipitation; Combined cross-linked enzyme aggregates (combi-CLEAs); Cross-Linking Reagents - chemistry; Fundamental and applied biological sciences. Psychology; Glucosidases - chemistry; Glucosidases - metabolism; Glucosyltransferases - chemistry; Glucosyltransferases - metabolism; Glutaral - chemistry; Maltooligosyltrehalose synthase; Maltooligosyltrehalose trehalohydrolase; Methods. Procedures. Technologies; Serum Albumin, Bovine; Sucrose - metabolism; Trehalose; Trehalose - metabolism; Maltooligosyltrehalose synthase; Trehalose; Amylosucrase; Combined cross-linked enzyme aggregates (combi-CLEAs); Maltooligosyltrehalose trehalohydrolase; Sucrose; Enzyme; Transferases; Glycosyltransferases; Synthase; Crosslinked state; Sequential; Biotransformation; Hexosyltransferases; Aggregate; Enzymatic reaction
• ISSN: 0960-8524; 1873-2976
• DOI: 10.1016/j.biortech.2012.12.162

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► Enzymatic one-pot bioconversion of sucrose to trehalose by combi-CLEAs. ► Development of combi-CLEAs using three enzymes, DGAS, BvMTS, and BvMTH. ► Optimization of various factors for the generation of combi-CLEAs. ► Reusability of DGAS–BvMTS–BvMTH combi-CLEAs was determined.
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.