Purification and characterization of d-allulose 3-epimerase derived from Arthrobacter globiformis M30, a GRAS microorganism

详细信息    查看全文

• 作者：Akihide Yoshihara¹ ; Taro Kozakai² ; Tomoya Shintani³ ; Ryo Matsutani³ ; Kouhei Ohtani³ ; Tetsuo Iida³ ; Kazuya Akimitsu² ; Ken Izumori² ; Pushpa Kiran Gullapalli³ ; ^{pushpa-kiran-gullapalli@matsutani.co.jp}
• 关键词：d-Allulose 3-epimerase ; Arthrobacter globiformis M30 ; d-Allulose ; Magnesium ; Immobilized d-AE
• 刊名：Journal of Bioscience and Bioengineering
• 出版年：2017
• 出版时间：February 2017
• 年：2017
• 卷：123
• 期：2
• 页码：170-176
• 全文大小：1074 K
• 卷排序：123

文摘

An enzyme that catalyzes C-3 epimerization between d-fructose and d-allulose was found in Arthrobacter globiformis strain M30. Arthrobacter species have long been used in the food industry and are well-known for their high degree of safety. The enzyme was purified by ion exchange and hydrophobic interaction chromatographies and characterized as a d-allulose 3-epimerase (d-AE). The molecular weight of the purified enzyme was estimated to be 128 kDa with four identical subunits. The enzyme showed maximal activity and thermostability in the presence of Mg2+. The optimal pH and temperature for enzymatic activity were 7.0–8.0 and 70°C, respectively. The enzyme was immobilized to ion exchange resin whereupon it was stable for longer periods than the free enzyme when stored at below 10°C. In the column reaction, the enzyme activity also maintained stability for more than 4 months. Under these conditions, 215 kg of d-allulose produced per liter immobilized enzyme, and this was the highest production yield of d-allulose reported so far. These highly stable properties suggest that this enzyme represents an ideal candidate for the industrial production of d-allulose.