*4.1. Materials*

Sodium alginate (*Macrocystis pyrifera* origin) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Alginate-assimilating bacteria, *Flavobacterium* sp. strain UMI-01, was cultivated at 25 ◦C in a mineral salt (MS) medium including 1% (*w*/*v*) sodium alginate as described in our previous report [26]. Cell lysate (crude enzyme) of this strain was extracted from cell pellets by freeze and thaw followed by sonication as described previously [28]. DEH was prepared by the digestion of sodium alginate with the crude enzyme and purified by SuperQ-650S (Tosoh, Tokyo, Japan) anion-exchange chromatography [28]. Standard KDG, KDPG, pyruvate, and GAP were purchased from Sigma-Aldrich. pCold I expression vector was purchased from TaKaRa (Shiga, Japan) and modified to the form that can add 8 × Gly + 8 × His-tag to the C-terminus of the expressed proteins [26]. *E. coli* DH5α and BL21 (DE3) were purchased from TaKaRa. Ni-NTA resin was purchased from Qiagen (Hilden, Germany). A TLC silica gel 60 plate was purchased from Merk KGaA (Darmstadt, Germany). TSKgel DEAE-2SW (4.6 × 250 mm) and Superdex peptide 10/300 GL were purchased from Tosoh Bioscience LLC (King of Prussia, PA, USA) and GE Healthcare (Little Chalfont, Buckinghamshire, UK), respectively. Lactate dehydrogenase (LDH; porcine heart origin) and NADH were purchased from Oriental Yeast Co., LTD. (Tokyo, Japan). ATP and 9-aminoacridine were purchased from Sigma-Aldrich. Other chemicals were purchased from Wako Pure Chemical Industries Ltd. (Osaka, Japan).
