Transformation of Total Notoginsenosides by Recombinant Endocellulase Fpendo5A†

doi:10.7503/cjcu20170314

Abstract

Abstract:

A novel endocellulase(Fpendo5A) was cloned from Fervidobacterium pennivorans DSM9078. After being overexpressed and purified from Escherichia coli, the enzymatic properties of Fpendo5A were investigated. Also, rapid resolution liquid chromatography coupled with quadruple-time-of-flight mass spectrometry(RRLC/Q-TOF-MS) was performed to investigate the biotransformation process of the total notoginsenosides by Fpendo5A. Fpendo5A exhibited an optimal activity at 80 ℃ and pH=5.5 and showed the highest activity for carboxymethyl cellulose(CMC), which indicated that Fpendo5A was a thermophilic and acidophi-lic endocellulase. Also, Fpendo5A showed high biotransformation ability for ginsenoside Ra₁, Rb₁, Rc, Rd and Rg₃, which are the main components in the total notoginsenosides. Ra₁, Rb₁, Rc, Rd and Rg₃ were converted to Rb₂, Gyp ⅩⅦ, CMC₁, F₂ and Rh₂, respectively by Fpendo5A.

Key words: Notoginsenoside, Endocellulase, Biotransformation, High performance liquid chromatography-mass spectrometry(HPLC-MS)

CLC Number:

O629
Q556

TrendMD:

LI Nan, ZHAO Huanxi, LI Jing, WANG Nan, YU Bohao, YUE Hao, YU Shanshan. Transformation of Total Notoginsenosides by Recombinant Endocellulase Fpendo5A^†[J]. Chem. J. Chinese Universities, 2017, 38(12): 2185.

Figures/Tables 5

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Peak No.	Compd.	t_R/min	Molecular formula	MS(calcd. mass), m/z	10⁶ Error	MS/MS fragment ions of [M-H]^-(mass accuracy < 5×10^-6)
1	Ra₁	21.74	C₅₈H₉₈O₂₆	1209.6242 (1209.6274)	2.7	1077.5839[M-H-Xyl ]^-, 945.5418[M-H-Xyl-Ara(p)]^-, 783.4874[M-H-Xyl-Ara-Glu]^-, 621.4336[M-H-Xyl-Ara-GlcGlc]^-, 459.5418[M-H-Xyl-Ara-GlcGlcGlc]^-, 323.0862[GlcGlc-H]^-
2	Rb₁	22.67	C₅₄H₉₂O₂₃	1107.5957 (1107.5976)	1.7	945.5294[M-H-Glc]^-, 783.4787[M-H-GlcGlc]^-,621.4295[M-H-GlcGlcGlc]^-, 459.33790[M-H-GlcGlcGlcGlc]^-, 323.0832[GlcGlc-H]^-
3	Rc	24.01	C₅₃H₉₀O₂₂	1077.5851 (1077.5875)	2.2	945.5193[M-H-Ara(f)]^-, 783.4760[M-H-Ara-Glc]^-, 621.4169[M-H-Ara-GlcGlc]^-, 459.3660[M-H-Ara-GlcGlcGlc]^-
4	Rb₂	24.93	C₅₃H₉₀O₂₂	1077.5851 (1077.5812)	3.6	945.5348[M-H-Ara(p)]^-, 783.4898[M-H-Ara-Glc]^-, 621.4365[M-H-Ara-GlcGlc]^-, 459.3852[M-H-Ara-GlcGlcGlc]^-
5	Rd	25.76	C₄₈H₈₂O₁₈	945.5428 (945.5462)	3.6	783.7832[M-H-Glc]^-, 621.5318[M-H-GlcGlc]^-, 459.5767[M-H-GlcGlcGlc]^-, 323.3125[GlcGlc-H]^-
6	Gyp ⅩⅦ	26.57	C₄₈H₈₂O₁₈	945.5428 (945.5437)	0.95	783.4615[M-H-Glc]^-, 621.4111[M-H-GlcGlc]^-, 459.9876[M-H-GlcGlcGlc]^-
7	CMC₁	28.28	C₄₇H₈₀O₁₇	915.5323 (915.5348)	2.7	783.1984[M-H-Ara(f)]^-, 621.6952[M-H-Ara-Glc]^-, 459.4832[M-H-Ara-GlcGlc]^-
8	F₂	29.11	C₄₂H₇₂O₁₃	783.4900 (783.4870)	3.8	621.3217[M-H-Glc]^-, 459.1521[M-H-GlcGlc]^-
9	Rg₃	30.83	C₄₂H₇₂O₁₃	783.4900 (783.4933)	4.2	621.4223[M-H-Glc]^-, 459.3791[M-H-GlcGlc]^-
10	Rh₂	33.66	C₃₆H₆₂O₈	621.4372 (621.4347)	4.0	459.3563[M-H-Glc]^-

Peak No.	Compd.	t_R/min	Molecular formula	MS(calcd. mass), m/z	10⁶ Error	MS/MS fragment ions of [M-H]^-(mass accuracy < 5×10^-6)
1	Ra₁	21.74	C₅₈H₉₈O₂₆	1209.6242 (1209.6274)	2.7	1077.5839[M-H-Xyl ]^-, 945.5418[M-H-Xyl-Ara(p)]^-, 783.4874[M-H-Xyl-Ara-Glu]^-, 621.4336[M-H-Xyl-Ara-GlcGlc]^-, 459.5418[M-H-Xyl-Ara-GlcGlcGlc]^-, 323.0862[GlcGlc-H]^-
2	Rb₁	22.67	C₅₄H₉₂O₂₃	1107.5957 (1107.5976)	1.7	945.5294[M-H-Glc]^-, 783.4787[M-H-GlcGlc]^-,621.4295[M-H-GlcGlcGlc]^-, 459.33790[M-H-GlcGlcGlcGlc]^-, 323.0832[GlcGlc-H]^-
3	Rc	24.01	C₅₃H₉₀O₂₂	1077.5851 (1077.5875)	2.2	945.5193[M-H-Ara(f)]^-, 783.4760[M-H-Ara-Glc]^-, 621.4169[M-H-Ara-GlcGlc]^-, 459.3660[M-H-Ara-GlcGlcGlc]^-
4	Rb₂	24.93	C₅₃H₉₀O₂₂	1077.5851 (1077.5812)	3.6	945.5348[M-H-Ara(p)]^-, 783.4898[M-H-Ara-Glc]^-, 621.4365[M-H-Ara-GlcGlc]^-, 459.3852[M-H-Ara-GlcGlcGlc]^-
5	Rd	25.76	C₄₈H₈₂O₁₈	945.5428 (945.5462)	3.6	783.7832[M-H-Glc]^-, 621.5318[M-H-GlcGlc]^-, 459.5767[M-H-GlcGlcGlc]^-, 323.3125[GlcGlc-H]^-
6	Gyp ⅩⅦ	26.57	C₄₈H₈₂O₁₈	945.5428 (945.5437)	0.95	783.4615[M-H-Glc]^-, 621.4111[M-H-GlcGlc]^-, 459.9876[M-H-GlcGlcGlc]^-
7	CMC₁	28.28	C₄₇H₈₀O₁₇	915.5323 (915.5348)	2.7	783.1984[M-H-Ara(f)]^-, 621.6952[M-H-Ara-Glc]^-, 459.4832[M-H-Ara-GlcGlc]^-
8	F₂	29.11	C₄₂H₇₂O₁₃	783.4900 (783.4870)	3.8	621.3217[M-H-Glc]^-, 459.1521[M-H-GlcGlc]^-
9	Rg₃	30.83	C₄₂H₇₂O₁₃	783.4900 (783.4933)	4.2	621.4223[M-H-Glc]^-, 459.3791[M-H-GlcGlc]^-
10	Rh₂	33.66	C₃₆H₆₂O₈	621.4372 (621.4347)	4.0	459.3563[M-H-Glc]^-