초록
<P><B>Abstract</B></P> <P>Trehalose 6-phosphate (Tre6<I>P</I>) is an important intermediate for trehalose biosynthesis. Recent researches have revealed that Tre6<I>P</I> is an endogenous signaling molecule that regulates plant development and stress responses. The necessity of Tre6<I>P</I> in physiological studies is expected to be increasing. To achieve the cost-effective production of Tre6<I>P,</I> a novel approach is required. In this study, we utilized trehalose 6-phosphate phosphorylase (TrePP) from <I>Lactococcus lactis</I> to produce Tre6<I>P</I>. In the reverse phosphorolysis by the TrePP, 91.9 mM Tre6<I>P</I> was produced from 100 mM β-glucose 1-phosphate (β-Glc1<I>P</I>) and 100 mM glucose 6-phosphate (Glc6<I>P</I>). The one-pot reaction of TrePP and maltose phosphorylase (MP) enabled production of 65 mM Tre6<I>P</I> from 100 mM maltose, 100 mM Glc6<I>P</I>, and 20 mM inorganic phosphate. Addition of β-phosphoglucomutase to this reaction produced Glc6<I>P</I> from β-Glc1<I>P</I> and thus reduced requirement of Glc6<I>P</I> as a starting material. Within the range of 20–469 mM inorganic phosphate tested, the 54 mM concentration yielded the highest amount of Tre6<I>P</I> (33 mM). Addition of yeast increased the yield because of its glucose consumption. Finally, from 100 mmol maltose and 60 mmol inorganic phosphate, we successfully achieved production of 37.5 mmol Tre6<I>P</I> in a one-pot reaction (100 mL), and 9.4 g Tre6<I>P</I> dipotassium salt was obtained.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Tre6<I>P</I> was formed from β-Glc1<I>P</I> and Glc6<I>P</I> by TrePP. </LI> <LI> With two other enzymes, Tre6<I>P</I> was produced from only maltose in one-pot reactions. </LI> <LI> Gram-scale synthesis of Tre6<I>P</I> from maltose and inorganic phosphate was established. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>