Ide Rb1, Rb2, Rc, and Rd. Rb1, Rb2, Rc, Rd, FIde Rb1, Rb2, Rc, and

December 27, 2023

Ide Rb1, Rb2, Rc, and Rd. Rb1, Rb2, Rc, Rd, F
Ide Rb1, Rb2, Rc, and Rd. Rb1, Rb2, Rc, Rd, F2, C-K, C-Y, and C-O, C-Mc, and C-Mc1, common ginsenosides. 1e3, enzyme reaction product from 25mM Rb1, 25mM Rb2, 25mM Rc reacted at 45 C for three h; four, enzyme reaction product from 2.5mM Rd. Solvent, chloroform:methanol:water sirtuininhibitor7.five:two.five:0.5; ten H2SO4 as a chromogenic agent.shown). Within the GRO-beta/CXCL2 Protein Storage & Stability purification, the yield in the ginsenosidase was about three.1 , as well as the precise activity on the enzyme improved 13 times (information not shown). three.two. Pure enzyme hydrolysis of the monomer ginsenosides Rb1, Rb2, Rc, and Rd The enzyme from the A. niger g.848 strain reacted with 25mM monomer ginsenoside Rb1, Rb2 and Rc at 45 C for three h, respectively; and reacted with two.5mM Rd at 45 C for 0.5 h. The enzyme reaction goods were examined by TLC (Fig. 2). As shown in Fig. two, the ginsenosidase produced by A. niger g.848 strain firstly hydrolyzed the 20-O-b-D-(1/6)-glucopyranoside of Rb1 into Rd, then hydrolyzed the 3-O-b-D-(1/2)-glucopyranoside of Rd to F2, further to C-K. Nevertheless, the enzyme firstly hydrolyzed the 3-O-b-D-(1/2)-glucopyranoside of Rb2 into C-O, hydrolyzed 3-O-b-D-glucopyranoside of C-O into C-Y, further hydrolyzed theAB0.40 0.30 AU 0.20 0.ten 0.00 two.00 4.6.six.8.00 Minutes10.12.14.Fig. 1. Purified ginsenosidase Type-I in SDS-PAGE and HPLC. (A) Ginsenosidase Type-I SDS-PAGE; marker, marker protein: phosphorylase b (97.2 kDa), serum albumin (66.four kDa), ovalbumin (44.3 kDa), carbonic anhydrase (29.0 kDa), trypsin inhibitor (20.1 kDa), and lysozyme (14.3 kDa). Protein quantity, 3 mg. (B) Ginsenosidase Type-I HPLC. HPLC, high performance liquid chromatography; Page, polyacrylamide gel electrophoresis; SDS, sodium dodecyl sulfate.C.-Y. Liu et al / Minor ginsenoside Neurofilament light polypeptide/NEFL Protein Species preparation20-O-a-L-(1/6)-arabinopyranoside (arap) of C-Y into C-K; the enzyme also firstly hydrolyzed the 3-O-b-D-(1/2)-glucopyranoside of Rc to C-Mc1, hydrolyzed the 3-O-b-D-glucopyranoside of CMc1 into C-Mc, and additional hydrolyzed 20-O-a-L-(1/6)-arabinofuranoside (araf) of C-Mc into C-K. The biotransformation pathway with the ginsenosidase developed by A. niger g.848 strain is shown in Fig. three. Thus, the enzyme from A. niger g.848 strain can hydrolyze the 3-C position (3-O-) and 20-C-position (20-O-) multiglycoside of PPD-type ginsenosides for instance Rb1, Rb2, Rc, and Rd, and should be classified to ginsenosidase type-I produced by Aspergillus sp.48 strain [23] along with a. niger g.48 strain [24]. However, the hydrolysis pathway on the specific ginsenosidase type-I from A. niger g.848 strain in present study is unique with that of ginsenosidase type-I from A. niger g.48 strain; the ginsenosidase type-I (molecular weight, 75 kDa) hydrolysis pathway from A. niger g.848 strain on ginsenoside Rb1 (in this study) is Rb1/Rd/F2/C-K; but the ginsenosidase type-I (molecular weight, 74 kDa) from A. niger g.48 strain hydrolyzes both 3-O- and 20-O-glucosides of Rb1 with two pathways: i.e., 1 pathway was Rb1/Rd/F2/C-K; another, Rb1/Gyp17/Gyp75/C-K [24]. Thus, the enzyme A. niger g.848 strain is actually a unique ginsenosidase type-I differentiating together with the ginsenosidase type-I from A. niger g.48 strain. The ginsenosidase type-I from A. niger g.848 strain is are suitable than that of A. niger g.48 strain, mainly because the ginsenoside Rb1 is hydrolyzed by ginsenosidase type-I from A. niger g.848 strain with a single pathway, and also the enzyme from A. niger g.48 strain hydrolyzes Rb1 with two pathways [24].3.three. Enzyme reaction kinetics To figure out the enzyme kinetic parameter.