灵芝子实体、菌丝体和孢子黑色素的结构表征与理化性质研究

doi:10.13560/j.cnki.biotech.bull.1985.2021-0659

摘要/Abstract

摘要：

为探讨灵芝子实体（GLM）、菌丝体（GLHM）和孢子（GLSM）黑色素结构和理化性质的差异,通过元素分析、紫外-可见光谱和傅里叶红外光谱等手段分析这3种黑色素的结构,并对这3种黑色素的理化性质如溶解性、色价、稳定性及体外抗氧化活性和对胰脂肪酶活性的抑制作用进行了比较研究。结果表明,这3种黑色素为真黑色素。与灵芝子实体和孢子黑色素相比,灵芝菌丝体黑色素具有更好的溶解性、色价和稳定性以及具有较强的体外抗氧化活性和体外对胰脂肪酶活性的抑制作用。研究结果为灵芝菌丝体黑色素的生物活性和实际应用提供理论基础。

关键词: 灵芝, 黑色素, 菌丝体, 体外抗氧化活性

Abstract:

In order to explore the difference in structure and physicochemical properties of melanin in fruiting body（GLM）,hyphae（GLHM）and spores（GLSM）of G. lucidum,three melanin’s structures were analyzed by elemental analysis,UV-Vis and FT-IR,and the physicochemical properties including solubility,color value and stability,antioxidant activity and inhibitory effect on pancreatic lipase activity in vitro were compared. The results showed that three melanin were eumelanin. Compared with GLM and GLSM,GLHM presented better solubility,color value and stability,as well as stronger antioxidant activity and stronger inhibitory effect on pancreatic lipase activity in vitro. This study will provide a theoretical basis for the bioactivity and practical application of melanin from G. lucidum hyphae.

Key words: Ganoderma lucidum, melanin, hyphae, antioxidant activity in vitro

王昭玉, 常明昌, 徐丽婧, 孟俊龙, 左宁柯, 潘旭. 灵芝子实体、菌丝体和孢子黑色素的结构表征与理化性质研究[J]. 生物技术通报, 2021, 37(11): 81-91.

WANG Zhao-yu, CHANG Ming-chang, XU Li-jing, MENG Jun-long, ZUO Ning-ke, PAN Xu. Structural Characterization,Physicochemical Properties of Melanin from Fruiting Body,Hyphae and Spores of Ganoderma lucidum[J]. Biotechnology Bulletin, 2021, 37(11): 81-91.

图/表 9

图1 同源灵芝子实体、菌丝体和孢子黑色素 A:灵芝子实体黑色素;B:灵芝菌丝体黑色素;C:灵芝孢子黑色素。下同

Fig. 1 Melanin of fruiting body,hyphae and spores of homologous G. lucidum A:Melanin in G. lucidum fruiting body（GLM）. B:Melanin in G. lucidum hyphae（GLHM）. C:Melanin in G. lucidum spores（GLSM）. The same below

表1 灵芝子实体、菌丝体和孢子黑色素的元素组成分析

Table 1 Elemental composition analysis of GLM,GLHM and GLSM

来源Source	元素组成Element composition/%
来源Source	C	H	N	O	S	C/N	C/H	O/N	S/N
灵芝子实体黑色素GLM	53.97	5.28	2.65	37.60	0.50	20.36	10.22	14.18	0.013
灵芝菌丝体黑色素GLHM	56.97	5.66	4.53	32.33	0.51	12.58	10.06	7.13	0.015
灵芝孢子黑色素GLSM	50.32	5.23	7.39	36.43	0.63	6.8	9.62	4.92	0.017
合成真黑色素Synthetic DOPA melanin	56.45	3.15	8.49	31.82	0.09	6.64	17.92	3.75	0.010
棕黑色素Phaeomelanin	46.24	4.46	9.36	30.16	9.78	4.94	10.37	3.22	1.040

图2 灵芝子实体、菌丝体和孢子黑色素的紫外可见光谱图

Fig. 2 UV-visible absorption spectra of GLM,GLHM and GLSM

图3 灵芝子实体、菌丝体和孢子黑色素的傅里叶红外光谱图

Fig. 3 FTIR spectra of GLM,GLHM and GLSM

表2 灵芝子实体、菌丝体和孢子黑色素的溶解性

Table 2 Solubility of GLM,GLHM and GLSM

溶液Solution	灵芝子实体黑色素GLM	灵芝菌丝体黑色素GLHM	灵芝孢子黑色素GLSM
1 mol·L^-1 HCl	-	-	-
1 mol·L^-1 NaOH	0.897±0.002	0.884±0.004	0.861±0.005
1 mol·L^-1 NaCl	-	-	-
蒸馏水Distilled water	-	-	-
正丁醇N-butanol	-	-	-
无水乙醇Absolute alcohol	0.009±0.001	-	-
75%无水乙醇75% absolute alcohol	0.015±0.002	0.016±0.001	-
异丙醇Isopropanol	-	-	-
氯仿Chloroform	-	-	-
乙酸乙酯Ethyl acetate	-	-	-

图4 灵芝子实体、菌丝体和孢子黑色素的温度稳定性和pH稳定性 A:灵芝子实体黑色素的温度稳定性;B:灵芝菌丝体黑色素的温度稳定性;C:灵芝孢子黑色素的温度稳定性;D:灵芝子实体、菌丝体和孢子黑色素的pH稳定性。图中误差线表示标准偏差。下同

Fig. 4 Thermal and pH stability of GLM,GLHM and GLSM A:Thermal stability of GLM. B:Thermal stability of GLHM. C:Thermal stability of GLSM. D:pH stability of GLM,GLHM and GLSM. The error line in the figure refers to the standard deviation. The same below

图5 灵芝子实体、菌丝体和孢子黑色素的光照稳定性和氧化还原性 A:灵芝子实体黑色素的光照稳定性;B:灵芝菌丝体黑色素的光照稳定性;C:灵芝孢子黑色素的光照稳定性;D:灵芝子实体、菌丝体和孢子黑色素的H2O2稳定性;E:灵芝子实体、菌丝体和孢子黑色素的Na2SO3稳定性

Fig. 5 Light stability and oxidation reduction of GLM,GLHM and GLSM A:Light stability of GLM. B:Light stability of GLHM. C:Light stability of GLSM. D:Stability of H2O2 in GLM,GLHM and GLSM. E:Stability of Na2SO3 in GLM,GLHM and GLSM

图6 灵芝子实体、菌丝体和孢子黑色素的体外抗氧化活性 A:灵芝子实体、菌丝体和孢子黑色素对DPPH的清除能力;B:灵芝子实体、菌丝体和孢子黑色素的还原能力;C:灵芝子实体、菌丝体和孢子黑色素超氧根阴离子清除能力;D:灵芝子实体、菌丝体和孢子黑色素羟基自由基的清除能力。小写字母表示的是同一浓度下Vc、灵芝子实体黑色素、灵芝菌丝体黑色素和孢子黑色素间差异达到了（P<0.05）显著水平,下同

Fig. 6 Antioxidant activities of GLM,GLHM and GLSM in vitro A:DPPH scavenging capacity of GLM,GLHM and GLSM. B:Reducing ability of GLM,GLHM and GLSM. C:Superoxide anion radical-scavenging ability of GLM,GLHM and GLSM. D:Hydroxyl radical scavenging ability of GLM,GLHM and GLSM. The lowercase letters indicate that the difference among VC,GLM,GLHM and GLSM at the same concentration reached a significant level（P< 0.05）. The same below

图7 灵芝子实体、菌丝体和孢子黑色素的对体外胰脂肪酶活性的抑制率

Fig. 7 Inhibitory rate of GLM,GLHM and GLSM on pancreatic lipase activity in vitro

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