One-Pot Biosynthesis of High-Concentration α-Glucose 1-Phosphate from Starch by Sequential Addition of Three Hyperthermophilic Enzymes

Journal of Agricultural and Food Chemistry
Wei ZhouY-H Percival Zhang

Abstract

α-Glucose 1-phosphate (G1P) is synthesized from 5% (w/v) corn starch and 1 M phosphate mediated by α-glucan phosphorylase (αGP) from the thermophilic bacterium Thermotoga maritima at pH 7.2 and 70 °C. To increase G1P yield from corn starch containing branched amylopectin, a hyper-thermostable isoamylase from Sulfolobus tokodaii was added for simultaneous starch gelatinization and starch-debranching hydrolysis at 85 °C and pH 5.5 before αGP use. The pretreatment of isoamylase increased G1P titer from 120 mM to 170 mM. To increase maltose and maltotriose utilization, the third thermostable enzyme, 4-glucanotransferase (4GT) from Thermococcus litoralis, was added during the late stage of G1P biotransformation, further increasing G1P titer to 200 mM. This titer is the highest G1P level obtained on starch or its derived products (maltodextrin and soluble starch). This study suggests that in vitro multienzyme biotransformation has an advantage of great engineering flexibility in terms of space and time compared with microbial fermentation.

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Citations

Nov 12, 2016·Journal of Industrial Microbiology & Biotechnology·Yi-Heng Percival ZhangYanhe Ma
Apr 15, 2017·Biotechnology and Bioengineering·Chun YouYi-Heng Percival Zhang
Nov 5, 2019·Biotechnology Journal·Chao Zhong, Bernd Nidetzky
Jun 11, 2017·Applied and Environmental Microbiology·Tomoko FujisawaHaruyuki Atomi
Jul 26, 2018·Applied Microbiology and Biotechnology·Zorica UbiparipTom Desmet
Mar 3, 2017·Applied Microbiology and Biotechnology·Hui ChenY-H Percival Zhang

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