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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2022, Vol. 42 Issue (11): 88-98    DOI: 10.13523/j.cb.2209040
    
Isolation and Biological Activity of Melanin from Sunflower Seed Shell
LI Chao-feng1,2,WU Yu-ping1,LI Shuang-shuang1,XU Feng-mei1,ZHANG Xin-yi1,WANG Wei-zhong1,2,TANG Bo-ping1,2,**()
1 School of Wetlands, Yancheng Teachers University, Yancheng 224007, China
2 Jiangsu Key Laboratory of Coastal Wetland Bioresources and Environmental Protection, Yancheng Teachers University, Yancheng 224007, China
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Abstract  

Objective: Natural edible pigments with high safety and low toxicity usually possess various nutritional and pharmacological effects, and they have huge practical application value in the market. In this paper, the extraction condition, antioxidant activity, which included DPPH radical-scavenging activity, superoxide radical (O2-)-scavenging activity and reducing power, and metal chelating activity of melanin from sunflower seed shell were investigated in detail to provide theoretical basis for its future application. Methods: In this paper, the efficient preparation of melanin from sunflower seed shell was achieved using hot alkali extraction by single-factor experiments combined with response surface methodology (RSM), acid hydrolysis, washing with organic solvents, re-dissolution and re-precipitation. The antioxidant activity and adsorption activity were determined by nitrotetrazolium nlue chloride reduction method, spectrophotometry and static adsorption. Results: Melanin from sunflower seed shell was successfully prepared. The optimal conditions were determined as follows: NaOH concentration, 0.11 mol/L; liquid-to-solid ratio, 15.77∶1; extraction time, 181.16 min; and extraction temperature, 75.19℃. Under these conditions, the experimental yield of melanin was 2.95%, which was well matched with the value predicted by the developed model. The ability to scavenge DPPH radical, superoxide radical (O2-) and the reducing power of melanin from sunflower seed shell were stronger than those of synthetic melanin. Furthermore, the adsorption efficiency of melanin from sunflower seed shell to Pb2+, Cu2+ and Cr3+ were higher than that of synthetic melanin. Conclusion: Compared with synthetic melanin, natural melanin from sunflower seed shell could be used as colorant, antioxidant and adsorbent, and was more effective. Therefore, melanin from sunflower seed shell was a potential replacement of synthetic melanin.

Key words： Sunflower seed shell      Melanin      Extraction      Antioxidant activity      Absorption     
Received: 15 September 2022      Published: 07 December 2022
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LI Chao-feng
WU Yu-ping
LI Shuang-shuang
XU Feng-mei
ZHANG Xin-yi
WANG Wei-zhong
TANG Bo-ping
Cite this article:

LI Chao-feng, WU Yu-ping, LI Shuang-shuang, XU Feng-mei, ZHANG Xin-yi, WANG Wei-zhong, TANG Bo-ping. Isolation and Biological Activity of Melanin from Sunflower Seed Shell. China Biotechnology, 2022, 42(11): 88-98.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2209040     OR     https://manu60.magtech.com.cn/biotech/Y2022/V42/I11/88

Fig.1 Effect of NaOH concentration on melanin extraction Different lowercase letters on the column indicate significant differences between different NaOH concentration treatments in the same period (P<0.05)
Fig.2 Effect of liquid-to-solid ratio on melanin extraction Different lowercase letters on the column indicate significant differences between different solid to liquid ratio treatments in the same period (P<0.05)
Fig.3 Effect of temperature on melanin extraction Different lowercase letters on the column indicate significant differences between different temperature treatments in the same period (P<0.05)
Fig.4 Effect of time on melanin extraction Different lowercase letters on the column indicate significant differences between different time treatments in the same period (P<0.05)
试验号 X1NaOH浓度/(mol/L) X2温度/℃ X3时间/h X4料液比 得率/%
1 0.1 60 2 15∶1 2.18±0.02
2 0.1 75 2 10∶1 2.18±0.03
3 0.1 75 4 20∶1 2.37±0.05
4 0.1 75 3 15∶1 2.98±0.02
5 0.2 60 3 15∶1 2.42±0.03
6 0.1 75 3 15∶1 2.96±0.04
7 0.2 75 3 10∶1 2.32±0.02
8 0.1 90 2 15∶1 2.30±0.03
9 0.0 75 3 10∶1 2.18±0.03
10 0.0 75 2 15∶1 2.26±0.01
11 0.2 75 2 15∶1 2.25±0.02
12 0.2 90 3 15∶1 2.32±0.02
13 0.1 75 2 20∶1 2.35±0.02
14 0.1 60 3 20∶1 2.41±0.03
15 0.1 75 4 10∶1 2.21±0.02
16 0.0 60 3 15∶1 2.09±0.01
17 0.2 75 4 15∶1 2.47±0.01
18 0.1 75 3 15∶1 2.95±0.02
19 0.1 90 3 20∶1 2.41±0.01
20 0.1 75 3 15∶1 2.94±0.02
21 0.2 75 3 20∶1 2.55±0.02
22 0.0 90 3 15∶1 2.32±0.02
23 0.0 75 3 20∶1 2.33±0.01
24 0.1 90 4 15∶1 2.26±0.01
25 0.1 75 3 15∶1 2.97±0.00
26 0.0 75 4 15∶1 2.10±0.03
27 0.1 90 3 10∶1 2.23±0.01
28 0.1 60 4 15∶1 2.26±0.02
29 0.1 60 3 10∶1 2.22±0.02
Table 1 Box-Behnken experimental design and results
变异来源 平方和 自由度 均方 F值 P值 显著性
模型 2.14 14 0.15 451.00 <0.000 1 **
X1 0.092 1 0.092 271.27 <0.000 1 **
X2 5.633×10-3 1 5.633×10-3 16.63 0.001 1 **
X3 1.875×10-3 1 1.875×10-3 5.54 0.033 8 *
X4 0.097 1 0.097 286.99 <0.000 1 **
X1X2 0.027 1 0.027 80.38 <0.000 1 **
X1X3 0.036 1 0.036 106.59 <0.000 1 **
X1X4 1.600×10-3 1 1.600×10-3 4.72 0.047 4 *
X2X3 3.600×10-3 1 3.600×10-3 10.63 0.005 7 **
X2X4 2.500×10-5 1 2.500×10-3 0.074 0.789 8
X3X4 2.500×10-5 1 2.500×10-3 0.074 0.789 8
X 1 2 0.66 1 0.66 1 961.13 <0.000 1 **
X 2 2 0.77 1 0.77 2 263.04 <0.000 1 **
X 3 2 0.90 1 0.90 2 657.41 <0.000 1 **
X 4 2 0.59 1 0.59 1 738.04 <0.000 1 **
残差 4.742×10-3 14 3.387×10-4
失拟项 3.742×10-3 10 3.742×10-4 1.50 0.371 4
纯误差 1.000×10-3 4 2.500×10-4
总误差 2.14 28
Table 2 Analysis of variance for the established regression model
Fig.5 Response surface showing the effects of different extraction condition on the yield of melanin from sunflower seed shell (a)NaOH solution and temperature (b)NaOH solution and time (c)NaOH solution and solid to liquid ratio (d)Temperature and time (e)Temperature and solid to liquid ratio (f)Time and solid to liquid ratio
序号 分析 结果
1 颜色 黑色
2 水 不溶
3 盐酸溶液 不溶
4
 甲醇、乙醇、氯仿、乙酸乙酯、石油醚、
正丁醇、丙酮等有机溶剂		 不溶
5
 氨水、NaOH、KOH、NaHCO3、
Na2CO3等碱性溶液		 溶解
6 H2O2、NaOCl、KMnO4、K2Cr2O7溶液 脱色
7 FeCl3溶液 棕色絮状物
Table 3 Physicochemical properties of melanin from sunflower seed shell
Fig.6 UV-vis spectra of melanin from sunflower seed shell (a)UV-vis absorption spectrum of the melanin from sunflower seed shell (b)A plot of log of optical density against wavelength
Fig.7 DPPH scavenging activity of melanin from sunflower seed shell
Fig.8 Superoxide anion-scavenging activity of melanin from sunflower seed shell
Fig.9 Reducing power of melanin from sunflower seed shell
Fig.10 Removal efficiency of melanin from sunflower seed shell
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