Mannose m1

The P. pastoris X33 transformant bearing the MrMnP manganese peroxidase gene from Moniliophthora roreri (GenBank accession number: ESK95360.1) was constructed by our lab [29 (link)], and this construct was used for MrMnP production. The substrates carboxymethyl cellulose (CMC), locust bean gum, 2,2’-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,6-dimethylphenol (2,6-DMP), and the standard substances glucose (CE1), cellobiose (CE2) and mannose (M1) were purchased from Sigma-Aldrich (St. Louis, MO). The substrates lichenan, mannohexaose, and the standard substances cellotriose (CE3), cellotetraose (CE4), cellopentaose (CE5), cellohexaose (CE6), mannobiose (M2), mannotriose (M3), mannotetraose (M4), mannopentaose (M5) and mannohexaose (M6) were purchased from Megazyme (Bray, Wicklow, Ireland). The other chemicals used in this research are of analytical grade and commercially available.

The actinomycetes were obtained from Biotechnology Culture Collection (BTCC), Indonesian Institute of Sciences (LIPI). Streptomyces lividans 1326 (NBRC 15675) was purchased from National Institute of Technology and Evaluation (NITE, Chiba, Japan). Escherichia coli JM109 (Takara, Shiga, Japan) was used as the host strain for DNA manipulation. Locust bean gum (LBG) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Ivory nut and konjac glucomannan were purchased from Megazyme (Wicklow, Ireland). Porang potato was purchased from PT Ambico (Surabaya, East Java, Indonesia). Suweg potato was purchased from a traditional market in East Java, Indonesia. Copra cake and palm kernel cake were purchased from a traditional market in Lampung, Sumatra, Indonesia. Mannose (M1) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Mannobiose (M2), mannotriose (M3), mannotetraose (M4), mannopentaose (M5), and mannohexaose (M6) were purchased from Megazyme (Wicklow, Ireland).

Mannose (M₁) and glucose (G₁) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Xylopentaose (X₅), cellobiose (G₂), cellotetraose (G₄), mannobiose (M₂), mannotriose (M₃), mannotetraose (M₄), mannopentaose (M₅), mannohexaose (M₆), 6¹-α-d-galactosyl-mannotriose (GalM₃), and 6³,6⁴-α-d-galactosyl-mannopentaose (Gal₂M₅) were purchased from Megazyme. Konjac glucomannan digest and carob galactomannan digest were produced in-house using RiGH26 (7 (link)) in 10 mM sodium phosphate (pH 5.8). Reaction mixtures were incubated for 16 h at 37°C following removal of RiGH26 using a Vivaspin 20 filtration unit (10,000-molecular-weight-cutoff [MWCO] polyethersulfone [PES]; Sartorius) and carbohydrate lyophilization on an ALPHA 2-4 LD Plus freeze dryer (Christ, Germany). Acetylated galactoglucomannan (AcGGM) was produced in-house as described by La Rosa et al. (24 (link)).