D glucose

The bacterial strain Pseudomonasputida MnB1 [American Type Culture Collection (ATCC) no. 23483] was cultured in the Lept medium (0.5 g/L yeast extract (Ruji, Shanghai, China), 0.5 g/L Casamino Acids (Coolaber, Beijing, China), 5 mM D(+)-glucose (Macklin, Shanghai, China), 10 mM HEPES (N-2-hydroxyethylpiperazine-N’-2-ethanesulfonic acid, pH 7.5, GBCBIO, Guangzhou, China), 0.48 mM CaC1₂ (Macklin), 0.83 mM MgSO₄ (Macklin), 3.7 μM FeCl₃ (Macklin), and 1 mL of trace element solution (10 mg/L CuSO₄•5H₂O, 44 mg/L ZnSO₄•7H₂O, 20 mg CoCl₂•6H₂O and 13 mg/L Na₂MoO₄•2H₂O, (Macklin)) containing 1mM MnCl₂ (Aladdin, Shanghai, China) at 30 °C and shaken at 150 rpm for 5 d [48 (link)]. The suspensions were centrifuged at 8000× g for 20 m and the supernatant was discarded, sediments of bacteria and BMO were diluted three times with deionized water by means of centrifugation (20 m at 8000× g), then the mixture of precipitates were collected for freeze-drying to obtain the dried BMO sample.

Mouse was orally administrated with 6 mg/kg body weight d-glucose (Macklin, Shanghai, China) dissolved in saline. The blood glucose levels at indicated time points (0, 15, 30, 60, 90 and 120 min) were measured using a handheld blood glucometer (Sinocare, Changsha, China) through tail tips.

The precursors d(+)-glucose (C₆H₁₂O₆, 99%) and sulfur (S, 99.9%) for catalyst preparation were purchased from Macklin. NaOH (AR, 96%), tetracycline (TC, CP), l-histidine (C₆H₉N₃O₂, 99%) and potassium persulfate (K₂S₂O₈, 99.5%) were obtained from Aladdin. Hydrochloric acid (HCl, AR) and methanol (MeOH, 99.7%) were obtained from Xilong Scientific Co., Ltd 20 nm nanoMgO was obtained from Xianfeng Nanomaterials Technology Co., Ltd 50 nm nanoMgO was obtained from Macklin. All chemicals were used as received. All solutions were prepared with ultrapure water.

All of the chemicals were of analytical grade, unless otherwise stated. The L-lysine, D-glucose, NaCl, xanthan gum, and gum arabic were purchased from Macklin Biochemical Co., Ltd. (Shanghai, China). The acetonitrile and formic acid of HPLC grade were acquired from Merck (Darmstadt, Germany). A solid-phase extraction cartridge, Cleanert PEP-2 (200 mg/6 mL, Bonna-Agela Technologies Inc., Tianjin, China), was used for the purification of the MR products. The pyrraline (purity >99.99%) and 3-deoxyglucosone (purity >99.99%) were supplied by Toronto Research Chemicals (North York, Ontario, Canada). The starches were obtained from Ingredion Incorporated (Shanghai, China).

D-glucose (Glu, 99.5%), L-phenylalanine (Phe, 99%), β-cyclodextrin (β-CD, 98%), sodium bisulfite (NaHSO₃, AR), and formic acid (chromatographic grade) were purchased from Macklin (Shanghai, China). Ethanol, glycerol, and acetic acid were purchased from Damao Chemical Reagent Co., Ltd. (Tianjin, China). Methyl alcohol (chromatographic grade) was purchased from Merck (99.9%, Darmstadt, Germany). Ultrapure water (UPW) was used for all the experiments.

The growth profiling was performed in triplicate in 9 cm Petri dishes on MM with 1.5% Ultra-pure agarose (Invitrogen, Carlsbad, CA, USA) with one of the following carbon sources: 25 mM D-glucose (Cat. no. G6172, Macklin, Shanghai, China), D-fructose (Cat. no. D809612, Macklin), D-xylose (Cat. no. XBO998, Sangon Biotech, Shanghai, China), L-arabinose (Cat. no. L824031, Macklin), cellobiose (Cat. no. C6182, Macklin), 1% w/v microcrystalline cellulose from cotton linters (Cat. no. 435236, Sigma, Shanghai, China), or 1% w/v xylan (from corn cob) (Cat. no. X823251, Macklin), or without an added carbon source. For growth profiling plate cultures, the plates were inoculated with a ~4 mm² agar plug from the growing edge of the T. harzianum colony from a PDA plate, and then incubated for at least four days at 28 °C with 12 h light and 12 h dark. After 48 h, the colony diameters were measured, and after both 48 h and 96 h, photos of the colonies were taken. Table S3 contains the colony diameter measurements from the two repeat experiments. Two independent deletion strains were tested for growth on the carbon sources to confirm the reliability of attributing the observed phenotypes to the deleted gene. We selected one of the deletion strains to use in further studies.