Invertase from baker s yeast

NEB buffer 4 and Nt.BbvCI endonuclease were obtained from New England BioLabs (Ipswich, MA, USA), and the polymerase Klenow fragment exo- was purchased from Thermo Fisher Scientific Inc. (Waltham, MA, USA). Streptavidin-coated 96-well plates were purchased from BEAVER Nano-Technologies (Suzhou, China). 3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonic acid (CHAPS), tris(2-carboxyethyl)phosphine hydrochloride (TCEP), Tween 20, and invertase from baker’s yeast were purchased from Sigma-Aldrich (St. Louis, MO, USA). Ethylene glycol, bis(aminoethyl ether)-N, N, N’, N’ tetraacetic acid (EGTA), glycerol, sulfosuccinimidyl-4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (sulfo-SMCC), bovine serum albumin (BSA), deoxynucleotide triphosphate (dNTP) solution mixture, 40% acrylamide mix solution, ammonium persulfate (APS), and 1,2-bis(dimethylamino)-ethane (TEMED) were obtained from Sangon Biotechnology Co. Ltd. (Shanghai, China). All other chemicals were of analytical grade and were used as received without purification. All water used in this work was RNase-free.
The oligonucleotides used in this assay (Table 1) were synthesized and purified by Sangon Biotechnology Co. Ltd. (Shanghai, China). Buffer I (0.1 M NaCl, 0.1 M sodium phosphate buffer, pH 7.3, 0.05% Tween-20) was used throughout the experiment.

Samples prepared for amino acid analysis were also used to measure glucose and sucrose levels. The samples were diluted with water (100 μL) appropriately and added to two separate test tubes (13 × 100 mm). One test tube was filled with 20 μL of buffer 30 mM of acetic-Na [pH 5.5] containing invertase (Invertase from baker’s yeast; Sigma, Kawasaki, Japan, 14,504) and incubated at 37 °C for 15 min to allow sucrose hydrolysis. The other test tube was filled with 20 μL of buffer only. Thereafter, glucose was measured using a glucose measuring kit (Glucose C2; Wako Pure Chemical Industries, Ltd., Japan). A₅₀₅ was measured using a spectrophotometer (Smartspec plus, Bio-Rad Laboratories, Inc., Hercules, CA, USA). The difference in glucose levels between tubes with and without invertase was attributed to sucrose-derived glucose.