Supporting Information for Antioxidative effects and mechanism study of bioactive peptides from defatted walnut (Juglans regia L.) meal hydrolysate Jianyong Sheng a, Xiaoyu Yang a, Jitang Chen a, Tianhao Peng a, Xiquan Yin b, Wei Liu a, Ming Liang b*, Jiangling Wan a,* and Xiangliang Yang a a National Engineering Research Center for Nanomedicine, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, Hubei 430074, People s Republic of China b Joint Laboratory for The research of Modern Preparation Technology- Huazhong University of Science and Technology and Infinitus, Guangzhou, Guangdong 510665, People s Republic of China *Corresponding author. Ming Liang, Tel: +86-20-80734744. E-mail: Fiona.Liang@infinitus-int.com * Corresponding author. Jiangling Wan, Tel: +86-27-67849580; fax: +86-27- 87792234. E-mail: wanjl@hust.edu.cn Jianyong Sheng and Xiaoyu Yang contributed equally to this work.
Table S1. Information of peptides with highest abundance ranking top15 in the de novo sequencing results of DWMH1 No. Sequences pi 1 VEGNLQVLRPR 9.45 2 QLQVLRPR 12.00 3 QLPR 9.12 4 VNLVNQHKLPL 8.63 5 LGLLPSFSNAPR 9.48 6 LLANEPR 5.98 7 NLPLL 5.48 8 VVDSEGKWEL 4.18 9 VLRGDA 5.84 10 AVLDLSNHANQLD 4.41 11 LVAVTVEDEL 3.65 12 LSLLPSYQPTSP 5.43 13 QNGAGFEWV 4.07 14 LSTVNSQNLPLL 5.60 15 TLMMSEEL 3.87 1
Table S2. Information of peptides with highest confidence ranking top15 in the de novo sequencing results of DWMH1 No. Sequences pi 16 LAGNPHQQQQN 6.77 17 EAQFGQQHVSGAGQ 5.16 18 HNLDTQTESDV 4.01 19 SLLDTNNNANQLDQNPR 4.19 20 NTNNNANQLDQNPR 5.69 21 AGNDGFEYVTLK 4.35 22 WSVWEQELEDR 3.98 23 NSALYVPHWNLNAH 7.13 24 QQRQQQGL 9.62 25 QNPDDEFRPQGQ 4.04 26 AELQVVDHLGQTV 4.39 27 EQEEEESTGRMK 4.26 28 QQERHHGQQQ 6.63 29 TMFQQQSQHPAQPPR 9.28 30 WSVWEQEL 3.98 2
Table S3. Information of peptides with highest abundance ranking top15 in the de novo sequencing results of DWMH2 No. Sequences pi 1 VEGNLQVLRPR 9.45 31 DVARVALAP 5.75 32 EPDVLALLQG 3.66 4 VNLVNQHKLPL 8.63 5 LGLLPSFSNAPR 9.48 6 LLANEPR 5.98 7 NLPLL 5.48 8 VVDSEGKWEL 4.18 9 VLRGDA 5.84 11 LVAVTVEDEL 3.65 33 RLDWVEA 4.42 34 PEEVLALA 3.88 14 LSTVNSQNLPLL 5.60 35 QPDVTQVFEFQ 3.68 36 VQLVDDNGDNVFDWV 3.38 3
Table S4. Information of peptides with highest confidence ranking top15 in the de novo sequencing results of DWMH2 No. Sequences pi 16 LAGNPHQQQQN 6.77 18 HNLDTQTESDV 4.01 21 AGNDGFEYVTLK 4.35 26 AELQVVDHLGQTV 4.39 37 MMRPDEDEQEGAGRQ 4.19 38 DTNNNANQLDQNPR 4.01 39 LAGNPDDEFTKQGPSQEYEQH 4.15 40 DDDLREGQL 3.78 41 LRHRADTQTESDV 5.44 27 EQEEEESTGRMK 4.26 24 QQRQQQGL 9.62 42 SDALYVPHWARNAH 6.97 22 WSVWEQELEDR 3.98 43 LSTVNSQNLPLL 9.48 44 LCVQQSGSNLFSGFDVCFL 3.90 4
Figure S1. MALDI-TOF mass spectrometry of (A) DWMH1 and (B) DWMH2. 5
(A) (B) (C) (D) (E) (F) (G) (H) Figure S2. HPLC chromatograms of peptides selected from DWMH. (A) P1: VEGNLQVLRPR, (B) P16: LAGNPHQQQQN, (C) P18: HNLDTQTESDV, (D) P21: AGNDGFEYVTLK, (E) P22: WSVWEQELEDR, (F) P24: QQRQQQGL, (G) P26: AELQVVDHLGQTV, (H) P27: EQEEEESTGRMK. 6
#810 RT: 8.04 AV: 1 NL: 2.90E7 F: FTMS + p ESI Full ms [100.00-2000.00] 100 95 90 85 80 75 70 65 60 55 50 45 40 35 30 25 20 15 629.7866 10 656.2432 1258.5679 438.1669 709.3336 939.4263 5 571.2473 810.3824 z=3 1026.4594 158.9630 1141.4875 233.1156 320.1484 1312.4846 1398.2382 1510.4823 1586.6887 1678.7559 1777.3451 1914.3173 z=3 0 100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600 1700 1800 1900 2000 m/z (A) (B) (C) 12 Relative Abundance (D) (E) 7
#323 RT: 3.09 AV: 1 NL: 2.71E7 F: FTMS + p ESI Full ms [100.00-2000.00] 100 95 90 85 80 75 70 65 60 55 50 45 40 35 30 25 20 15 10 429.2386 475.7500 493.2681 985.5291 5 347.1543 158.9634 519.7245 652.3243 223.0667 z=3 780.3845 837.4070 968.5008 1012.4887 1102.6094 1230.2778 1314.0425 1478.2914 1552.4701 1686.4683 1825.3682 z=3 0 100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600 1700 1800 1900 2000 m/z #1921 RT: 13.07 AV: 1 NL: 6.22E7 F: FTMS + p ESI Full ms [100.00-2000.00] 100 95 90 85 80 75 70 65 60 55 50 45 40 35 30 25 20 113.9654 488.2328 z=3 704.8849 15 687.3215 470.2597 z=3 10 154.9926 209.4309 366.4265 z=4 731.8455 5 287.8933 506.2063 631.8045 379.9064 z=3 868.4648 940.1802 994.4321 1083.1677 1190.6348 1275.4946 1408.7648 1545.6146 1748.9945 1881.0602 z=4 z=3 z=3 0 100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600 1700 1800 1900 2000 m/z (F) 13 Relative Abundance (G) 14_170926092356 Relative Abundance (H) Figure S3. Mass spectrums of peptides selected from DWMH. (A) P1: VEGNLQVLRPR, (B) P16: LAGNPHQQQQN, (C) P18: HNLDTQTESDV, (D) P21: AGNDGFEYVTLK, (E) P22: WSVWEQELEDR, (F) P24: QQRQQQGL, (G) P26: AELQVVDHLGQTV, (H) P27: EQEEEESTGRMK. 8
Figure S4. The different concentration of H 2 O 2 induced oxidative damage in SH-SY5Y cells. * p<0.05 vs the control group. 9
Figure S5. Protective effects of different concentration of peptides derived from DWMH or DWMH on H 2 O 2 induced oxidative damage in SH-SY5Y cells. * means had significant difference with the 150 μg/ml group, # means had significant difference with the 300 μg/ml group. 10