Article(id=1320888982410473585, tenantId=1146029695717560320, journalId=1260987677001138203, issueId=1320888908896907909, articleNumber=null, orderNo=null, doi=10.13386/j.issn1002-0306.2025050071, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1746979200000, receivedDateStr=2025-05-12, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1792854074337, onlineDateStr=2026-10-24, pubDate=1772294400000, pubDateStr=2026-03-01, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1792854074337, onlineIssueDateStr=2026-10-24, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1792854074337, creator=13701087609, updateTime=1792854074337, updator=13701087609, issue=Issue{id=1320888908896907909, tenantId=1146029695717560320, journalId=1260987677001138203, year='2026', volume='47', issue='5', pageStart='1', pageEnd='503', issueExtLink='null', onlineDate='null', pubDate='1772294400000', pubDateStr='2026-03-01', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1792854056809, creator='13701087609', updateTime=1792854056809, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext=null, issueFiles=null, downloadFileDto=null}, startPage=428, endPage=434, ext={EN=ArticleExt(id=1320888982620188786, articleId=1320888982410473585, tenantId=1146029695717560320, journalId=1260987677001138203, language=EN, title=Comparative Study on Antioxidant Activities of Liquid Fermented and Fruiting Body Polysaccharides from Pleurotus ostreatus, columnId=1261343846965830077, journalTitle=Science and Technology of Food Industry, columnName=Nutrition and Healthcare, runingTitle=null, highlight=null, articleAbstract=

In order to study the difference of antioxidant activities among the extracellular polysaccharide (POFP), mycelium polysaccharide (POMP) and fruiting body polysaccharide (POP) of Pleurotus ostreatus by liquid fermentation, this study prepared POFP and POMP through liquid-state fermentation of edible fungi, polysaccharide isolation and purification, as well as other techniques, followed by a comparative evaluation of their antioxidant activities with POP. The study assessed the in vitro antioxidant activity of the three polysaccharides using DPPH radical, ABTS+ radical, and OH radical scavenging activities as well as total reducing power. Additionally, yeast cell models subjected to ultraviolet (UV)-induced oxidative damage and H2O2-induced oxidative damage were employed to evaluate their protective effects against oxidative injury. The results showed that the three kinds of polysaccharides were composed of two kinds of components with different molecular weights, which ranged from 375.31~9788.14 Da, and the order of molecular weight was POP>POMP>POFP. All three polysaccharides demonstrated significant scavenging activities against DPPH, ABTS+, and OH radicals, along with notable reducing power, all of which exhibited a dose-dependent manner. Among them, POFP displayed the strongest total reducing power, achieving an OD700 value of 0.51±0.04 at a concentration of 5 mg/mL, which was higher than that of POMP and POP. Furthermore, all three polysaccharides significantly increased the survival rates of yeast cells subjected to UV- and H2O2-induced oxidative damage. Among them, POP demonstrated stronger protective effects, restoring the viability of UV-damaged yeast cells to 96.05%±4.02% at a concentration of 25 mg/mL, and that of H2O2-damaged yeast cells to 91.6%±3.26% at 15 mg/mL, which was comparable to the effects observed in the vitamin C (VC) treatment group. In summary, Pleurotus ostreatus polysaccharides from different sources exhibited significant antioxidant activities, and liquid fermentation technology provided a new way for the preparation of new antioxidant natural products.

, authors=Wei ZHOU1, Hongjia SHEN1, Quan GAN1, Cuiying SONG1, Lei ZHANG2, Liming ZHANG1, Peipei HAN1, *, Limin HAO2, *, Shiru JIA1, authorsList=Wei ZHOU, Hongjia SHEN, Quan GAN, Cuiying SONG, Lei ZHANG, Liming ZHANG, Peipei HAN, Limin HAO, Shiru JIA, authorCompany=null, correspAuthors=Peipei HAN, Limin HAO, authorNote=null, correspAuthorsNote=null, copyrightStatement=Copyright © 2026 Science and Technology of Food Industry. All rights reserved., copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1320888985375846527, articleId=1320888982410473585, tenantId=1146029695717560320, journalId=1260987677001138203, language=CN, title=平菇液态发酵多糖与其子实体多糖抗氧化活性对比, columnId=1261343850979779009, journalTitle=食品工业科技, columnName=营养与保健, runingTitle=null, highlight=null, articleAbstract=

为研究平菇液态发酵胞外多糖(POFP)、平菇菌丝体多糖(POMP)和平菇子实体多糖(POP)的抗氧化活性差异,本研究采用食用菌液态发酵、多糖分离纯化等技术制备了POFP和POMP,并与POP进行抗氧化活性对比评价。研究采用DPPH自由基、ABTS+自由基、OH自由基清除能力和总还原力评价三种多糖的体外抗氧化活性,并采用紫外线氧化损伤和H2O2氧化损伤酵母细胞模型来评估其氧化损伤防护作用。结果表明:三种多糖均由两种不同分子量的组分构成,分子量在375.31~9788.14 Da之间,大小排序为POP>POMP>POFP;三种多糖对DPPH、ABTS+、OH自由基均有较好的清除能力,并且具有良好的还原能力,且呈现剂量依赖性。其中POFP的总还原力最强,在5 mg/mL时OD700值可达0.51±0.04,高于POMP和POP。此外,三种多糖均可显著提高紫外线和H2O2氧化损伤酵母细胞存活率;POP防护效果较强,在25 mg/mL时可使紫外线损伤酵母细胞活力恢复至96.05%±4.02%,在15 mg/mL可使H2O2氧化损伤酵母细胞活力恢复至91.60%±3.26%,与VC处理组效果相当。综上所述,不同来源的平菇多糖均具有良好的抗氧化活性,液态发酵技术为新型抗氧化天然产物的制备提供了新路径。

, authors=周威1, 申红佳1, 甘泉1, 宋翠颖1, 张蕾2, 张黎明1, 韩培培1, *, 郝利民2, *, 贾士儒1, authorsList=周威, 申红佳, 甘泉, 宋翠颖, 张蕾, 张黎明, 韩培培, 郝利民, 贾士儒, authorCompany=null, correspAuthors=韩培培, 郝利民, authorNote=

周威(1999−),男,硕士研究生,研究方向:生物工程,E-mail:

, correspAuthorsNote=
韩培培(1983−),女,博士,教授,研究方向:生物工程,E-mail:。
郝利民(1969−),男,博士,教授,研究方向:特殊功能食品与食品生物技术,E-mail:。
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注:不同大写字母表示同一浓度下不同来源平菇多糖的清除率存在显著差异(P<0.05),不同小写字母表示同一来源平菇多糖不同浓度的清除率存在显著差异(P<0.05),图2~图6同。

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Basic composition of POFP, POMP and POP

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指标POPPOFPPOMP
总糖含量(%)84.20±0.28a55.53±0.09b44.72±0.17c
蛋白质含量(%)10.84±0.29c14.50±0.90b27.00±0.12a
水分含量(%)3.21±0.46b5.18±0.08a1.34±0.22c
), ArticleFig(id=1320888991482753226, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888982410473585, language=CN, label=表1, caption=

POFP、POMP和POP的基本组成

, figureFileSmall=null, figureFileBig=null, tableContent=
指标POPPOFPPOMP
总糖含量(%)84.20±0.28a55.53±0.09b44.72±0.17c
蛋白质含量(%)10.84±0.29c14.50±0.90b27.00±0.12a
水分含量(%)3.21±0.46b5.18±0.08a1.34±0.22c
), ArticleFig(id=1320888991554056395, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888982410473585, language=EN, label=Table 2, caption=

Relative molecular weights of POFP, POMP and POP

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粗多糖保留时间(min)重均分子量Mw(Da)
POFP12.19375.31
12.01421.83
POMP9.901675.60
8.514157.11
POP7.756832.11
7.209788.14
), ArticleFig(id=1320888991612776652, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320888982410473585, language=CN, label=表2, caption=

POFP、POMP和POP的相对分子量

, figureFileSmall=null, figureFileBig=null, tableContent=
粗多糖保留时间(min)重均分子量Mw(Da)
POFP12.19375.31
12.01421.83
POMP9.901675.60
8.514157.11
POP7.756832.11
7.209788.14
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平菇液态发酵多糖与其子实体多糖抗氧化活性对比
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周威 1 , 申红佳 1 , 甘泉 1 , 宋翠颖 1 , 张蕾 2 , 张黎明 1 , 韩培培 1, * , 郝利民 2, * , 贾士儒 1
食品工业科技 | 营养与保健 2026,47(5): 428-434
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食品工业科技 |营养与保健 2026 , 47 (5) : 428 -434
平菇液态发酵多糖与其子实体多糖抗氧化活性对比
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周威1 , 申红佳1, 甘泉1, 宋翠颖1, 张蕾2, 张黎明1, 韩培培1, * , 郝利民2, * , 贾士儒1
作者信息
  • 1天津科技大学,工业发酵微生物教育部重点实验室,天津 300457
  • 2军事科学院系统工程研究院,北京 100010
通讯作者:
韩培培(1983−),女,博士,教授,研究方向:生物工程,E-mail:。
郝利民(1969−),男,博士,教授,研究方向:特殊功能食品与食品生物技术,E-mail:。
作者简介:

周威(1999−),男,硕士研究生,研究方向:生物工程,E-mail:

Comparative Study on Antioxidant Activities of Liquid Fermented and Fruiting Body Polysaccharides from Pleurotus ostreatus
Wei ZHOU1 , Hongjia SHEN1, Quan GAN1, Cuiying SONG1, Lei ZHANG2, Liming ZHANG1, Peipei HAN1, * , Limin HAO2, * , Shiru JIA1
Affiliations
  • 1Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin University of Science and Technology, Tianjin 300457, China
  • 2Systems Engineering Institute, Academy of Military Sciences (AMS), Beijing 100010, China
出版时间: 2026-03-01 doi: 10.13386/j.issn1002-0306.2025050071
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为研究平菇液态发酵胞外多糖(POFP)、平菇菌丝体多糖(POMP)和平菇子实体多糖(POP)的抗氧化活性差异,本研究采用食用菌液态发酵、多糖分离纯化等技术制备了POFP和POMP,并与POP进行抗氧化活性对比评价。研究采用DPPH自由基、ABTS+自由基、OH自由基清除能力和总还原力评价三种多糖的体外抗氧化活性,并采用紫外线氧化损伤和H2O2氧化损伤酵母细胞模型来评估其氧化损伤防护作用。结果表明:三种多糖均由两种不同分子量的组分构成,分子量在375.31~9788.14 Da之间,大小排序为POP>POMP>POFP;三种多糖对DPPH、ABTS+、OH自由基均有较好的清除能力,并且具有良好的还原能力,且呈现剂量依赖性。其中POFP的总还原力最强,在5 mg/mL时OD700值可达0.51±0.04,高于POMP和POP。此外,三种多糖均可显著提高紫外线和H2O2氧化损伤酵母细胞存活率;POP防护效果较强,在25 mg/mL时可使紫外线损伤酵母细胞活力恢复至96.05%±4.02%,在15 mg/mL可使H2O2氧化损伤酵母细胞活力恢复至91.60%±3.26%,与VC处理组效果相当。综上所述,不同来源的平菇多糖均具有良好的抗氧化活性,液态发酵技术为新型抗氧化天然产物的制备提供了新路径。

平菇  /  液态发酵  /  平菇多糖  /  抗氧化活性  /  氧化损伤

In order to study the difference of antioxidant activities among the extracellular polysaccharide (POFP), mycelium polysaccharide (POMP) and fruiting body polysaccharide (POP) of Pleurotus ostreatus by liquid fermentation, this study prepared POFP and POMP through liquid-state fermentation of edible fungi, polysaccharide isolation and purification, as well as other techniques, followed by a comparative evaluation of their antioxidant activities with POP. The study assessed the in vitro antioxidant activity of the three polysaccharides using DPPH radical, ABTS+ radical, and OH radical scavenging activities as well as total reducing power. Additionally, yeast cell models subjected to ultraviolet (UV)-induced oxidative damage and H2O2-induced oxidative damage were employed to evaluate their protective effects against oxidative injury. The results showed that the three kinds of polysaccharides were composed of two kinds of components with different molecular weights, which ranged from 375.31~9788.14 Da, and the order of molecular weight was POP>POMP>POFP. All three polysaccharides demonstrated significant scavenging activities against DPPH, ABTS+, and OH radicals, along with notable reducing power, all of which exhibited a dose-dependent manner. Among them, POFP displayed the strongest total reducing power, achieving an OD700 value of 0.51±0.04 at a concentration of 5 mg/mL, which was higher than that of POMP and POP. Furthermore, all three polysaccharides significantly increased the survival rates of yeast cells subjected to UV- and H2O2-induced oxidative damage. Among them, POP demonstrated stronger protective effects, restoring the viability of UV-damaged yeast cells to 96.05%±4.02% at a concentration of 25 mg/mL, and that of H2O2-damaged yeast cells to 91.6%±3.26% at 15 mg/mL, which was comparable to the effects observed in the vitamin C (VC) treatment group. In summary, Pleurotus ostreatus polysaccharides from different sources exhibited significant antioxidant activities, and liquid fermentation technology provided a new way for the preparation of new antioxidant natural products.

Pleurotus ostreatus  /  liquid fermentation  /  Pleurotus ostreatus polysaccharide  /  antioxidant activity  /  oxidative damage
周威, 申红佳, 甘泉, 宋翠颖, 张蕾, 张黎明, 韩培培, 郝利民, 贾士儒. 平菇液态发酵多糖与其子实体多糖抗氧化活性对比. 食品工业科技, 2026 , 47 (5) : 428 -434 . DOI: 10.13386/j.issn1002-0306.2025050071
Wei ZHOU, Hongjia SHEN, Quan GAN, Cuiying SONG, Lei ZHANG, Liming ZHANG, Peipei HAN, Limin HAO, Shiru JIA. Comparative Study on Antioxidant Activities of Liquid Fermented and Fruiting Body Polysaccharides from Pleurotus ostreatus[J]. Science and Technology of Food Industry, 2026 , 47 (5) : 428 -434 . DOI: 10.13386/j.issn1002-0306.2025050071
平菇(Pleurotus ostreatus)属担子菌门(Basidiomycotina)侧耳属(Pleurotus),又被称为北风菌、蚝菌,在我国食用菌产量排名中位居前列,是全球继双孢蘑菇之后的第二大栽培食用菌,具有重要的经济、生态及食药用价值[1−2]。平菇多糖是平菇中最主要的活性物质之一,具有抗疲劳、抗氧化、调节免疫、抗肿瘤、降血糖、降血脂、抗凝血和保肝护肝等作用,对机体健康有诸多益处[3]。平菇多糖通常分为子实体多糖和液态发酵多糖,其中子实体多糖从平菇子实体中提取获得,不同的提取方法会影响多糖的提取得率、结构特征以及生物活性。传统食用菌多糖提取法主要有热水浸提法和酸碱溶液提取法,近年超声辅助法、酶提取辅助法、双水相提取法等也逐渐兴起[4]。然而平菇子实体培养周期较长,一般为30~45 d,受天气和季节等自然因素影响较大,生产成本高、效率低;而且生产中存在原料提取率低、废渣大量浪费的现象[5−8]。平菇液态发酵培养可以缩短培养周期,减少人力投入,且不受季节和地域限制,规模化和自动化程度较高,发酵过程中的关键参数如温度和pH能够精确调控[9],从而保证发酵始终在最优状态下进行;此外,液态发酵技术简化了平菇活性物质提取步骤,提高了生产效率,是平菇多糖产业化制备的重要技术路径。
人体内氧化和抗氧化失衡会引发机体氧化应激,导致自由基和炎症因子的增加,进而对机体产生多方面的影响,例如加速机体衰老,提高慢病发生率等,对健康造成危害[10−12]。研究表明,平菇多糖具有良好的抗氧化活性,其作用机制主要是提高自由基清除能力或增强抗氧化酶活性[13]。程超等[14]研究发现平菇子实体多糖对超氧阴离子和OH自由基具有良好的清除能力;Li等[15]发现平菇多糖能够显著提高环磷酰胺(CTX)诱导的小鼠睾丸组织中核转录因子Nrf2及其下游ARE基因的表达,同时改变Keap1蛋白域结构,降低其表达,促进抗氧化酶的产生。董博斐等[16]研究表明,红平菇液体发酵胞外多糖对机体的氧化应激损伤具有良好的修复作用;曹向宇等[17]使用酶解的方法提取纯化了平菇胞内多糖,其对超氧阴离子和OH自由基均具有清除作用。虽然目前对平菇多糖的研究较多,但大部分集中在单一来源多糖的抗氧化活性研究,而对不同来源多糖例如子实体多糖和液态发酵多糖之间的抗氧化活性差异比较研究较少。
本文系统比较了平菇发酵液多糖(POFP)、平菇菌丝体多糖(POMP)与平菇子实体多糖(POP)的体外抗氧化活性差异,探究了三类多糖对DPPH、ABTS+、OH自由基的清除能力及对紫外线、H2O2氧化损伤酵母细胞的保护作用,为不同来源平菇多糖在功能性食品和美容护肤品领域的应用提供了参考。
平菇2180菌种 天津农学院农学与资源环境学院食用菌研发中心;平菇子实体 河南菇太郎食品有限公司;酿酒酵母A.S.2399 天津科技大学食品营养与安全国家重点实验室;1,1-二苯基-2-三硝基苯肼(DPPH)、2,2-联氮基双(3-乙基苯并噻唑啉-6-磺酸)二铵盐(ABTS)、牛血清蛋白、单糖标准样 美国Sigma公司;二喹啉甲酸(bicinchoninic acid,BCA)蛋白浓度测定试剂盒 上海阿拉丁试剂有限公司;无水乙醇、苯酚、浓硫酸、氯仿、正丁醇、氯化钠、三氟乙酸 均为国产分析纯。
LDZX-75L型立式高压蒸汽灭菌器 上海申安医疗器械厂;HYG型恒温摇床培养箱 上海欣蕊自动化设备有限公司;TGL16G型高速离心机 上海医用分析仪器厂;TDA-8002型恒温水浴锅 天津市中环试验电炉有限公司;ALPHA 1-2/LD型真空冷冻干燥机 德国CHRIS公司;JA12002型电子天平 上海精科天平仪器厂;RE-3000型旋转蒸发仪 上海亚荣生化仪器厂;Infinite M200 Pro型多功能酶标仪 瑞士Tecan公司;Aglient 1200液相色谱仪 德国安捷伦科技有限公司。
采用王菲菲等[18]的菌种发酵方法并稍作修改。通过前期试验得到最优液态培养基配方为:葡萄糖30.0 g/L、酵母粉8.5 g/L、KH2PO4 1.5 g/L、MgSO4·7H2O 0.5 g/L、VB1 0.1 g/L,pH自然。将活化好的平菇菌种接种5%于液态培养基中,在26 ℃、180 r/min条件下摇床培养5 d,4 ℃下10000 r/min离心处理5 min后获得平菇发酵液(POF)和菌丝体(POM),并通过5 L至500 L发酵罐进行逐级放大试验。
采用刘宇[19]的方法制备POFP和POMP。将POM用蒸馏水清洗干净后冷冻干燥,取一定量POM粉末进行研磨破壁处理,按料液比1:35(g/mL)热水浸提(90 ℃、2 h)2次,4 ℃下10000 r/min离心处理5 min后合并上清液,得POM提取液。将上述制得的POF和POM提取液分别浓缩至一定体积,加入3倍体积95%乙醇溶液,在4 ℃的环境下,醇沉12 h,8000 r/min离心5 min收集沉淀,用95%乙醇溶液进行多次冲洗,加适量水溶解后经过冷冻干燥后分别得到POFP和POMP。
将平菇子实体在60 ℃烘箱内烘干、粉碎[20],取一定量平菇子实体干粉,后续操作按照1.2.1中热水浸提方法提取子实体多糖POP。
总糖含量采用苯酚-硫酸法测定;蛋白含量采用BCA法测定;水分含量采用食用水分蒸发仪进行检测[21]。
POFP、POMP和POP多糖经过Sevage法除蛋白、AB-8大孔树脂脱色、真空浓缩,冷冻干燥后制得待测样品。采用高效凝胶过滤色谱法(High performance gel filtration chromatography,HPGFC)测定粗多糖相对分子质量。测试条件如下,色谱柱:TSK GEL4000SWXL(7.8 mm×30.0 cm);检测器:RI-101示差折光检测器;流动相:超纯水;流速:0.5 mL/min;柱温:35 ℃;将样品溶解在液相中,用0.22 μm微孔滤膜过滤,进样量为20 μL。葡聚糖标准曲线采用超纯水配制浓度为1 mg/mL的葡聚糖标准品溶液,并依照所述色谱分析条件进行检测。根据标准品葡聚糖分子量的对数值(lgMw)与保留时间(tR)进行线性回归分析,其回归方程为y=−0.2839x+6.03478,R2=0.9966,根据标准曲线计算平菇多糖样品分子质量。
参考Chen等[22]的方法测定DPPH自由基的清除能力,并稍作修改。配制不同浓度(1、2、3、4、5 mg/mL)的粗多糖溶液,取100 μL粗多糖溶液于96孔板中,加入100 μL DPPH溶液(0.1 mmol/L,溶剂为无水乙醇)后,在37 ℃的环境下,黑暗处理30 min,在517 nm处测定混合物的吸光度。其中,VC为阳性对照。DPPH自由基清除率根据公式(1)计算:
$\rm {DPPH自由基清除率}(\text{%})=\left[\frac{{{A}}_{{k}}-({{A}}_{{i}}-{{A}}_{{j}})}{{{A}}_{{k}}}\right]\times 100 $
式中,Ak表示空白组吸光度;Ai表示样品组吸光度;Aj表示样品本底吸光度。
按照Wang等[23]的方法进行测定。将浓度为7.4 mmol/L的ABTS+溶液、浓度为2.6 mmol/L的过硫酸钾溶液,等比例混匀后在黑暗条件下反应16 h,并用pH7.4的PBS缓冲液稀释,在734 nm下测吸光度,该吸光度在0.70±0.02范围内,制得ABTS+工作液。取30 μL不同浓度(1、2、3、4、5 mg/mL)多糖溶液于96孔板内,加入180 μL ABTS+工作液并混匀,室温下反应6 min,测定734 nm处的吸光度。以VC溶液作阳性对照。ABTS+自由基清除率根据公式(2)计算:
$ {\text{ABTS}}^{\text+}\text{自由基清除率}(\text{%})=\left[\frac{{\mathrm{A}}_{\mathrm{k}}-\left({\mathrm{A}}_{\mathrm{i}}-{\mathrm{A}}_{\mathrm{j}}\right)}{{\mathrm{A}}_{\mathrm{k}}}\right]\text{×100} $
式中,Ak表示空白组吸光度;Ai表示样品组吸光度;Aj表示样品本底吸光度。
根据Zhang等[24]的方法并稍作改进。配制不同浓度(1、2、3、4、5 mg/mL)的粗多糖溶液,依次在96孔板中加入50 μL的粗多糖溶液、5.0 mmol/L水杨酸-乙醇溶液、5.0 mmol/L FeSO4溶液和5.0 mmol/L H2O2溶液,充分混匀后在水浴锅中加热30 min,温度为37 ℃,并在510 nm处测定其吸光度。以VC为阳性对照。OH自由基清除率根据公式(3)计算:
$ \text{OH自由基清除率}(\text{%})=\left[\frac{\text{A}_{\text{k}}-\left(\text{A}_{\text{i}}-\text{A}_{\text{j}}\right)}{\text{A}_{\text{k}}}\right]\text{×100} $
式中,Ak表示空白组吸光度;Ai表示样品组吸光度;Aj表示样品本底吸光度。
参照Fan等[25]所述的方法进行铁还原抗氧化能力测定,并稍作修改。在试管内加入2 mL不同浓度(1、2、3、4、5 mg/mL)的多糖样品后,分别加入2 mL的1%的铁氰化钾溶液和pH6.6的PBS缓冲液,在水浴锅中加热20 min,温度为50 ℃,加入2 mL 10%三氯乙酸溶液,混匀后5000 r/min 离心10 min。取上清液100 μL于96孔板内,继续加入10 μL的0.1%三氯化铁溶液,混匀后50 ℃水浴10 min,待温度降至室温后,在700 nm下测定混合液的吸光度。以VC溶液作阳性对照。还原能力根据公式(4)计算:
$\rm {\text{还原能力}=A}_{\text{i}}-{A}_{\text{j}} $
式中,Ai表示样品组吸光度;Aj表示样品本底吸光度。
参照圣志存等[26]的方法进行测定,并稍加修改。将活化好的酵母细胞接种于YEPD液体培养基中,培养至对数期后,取适量酵母培养液4000 r/min离心5 min,弃上清,用生理盐水将菌体重复洗涤两次后制成酵母菌悬液。将不同质量浓度(5、10、15、20、25 mg/mL)的粗多糖样品与酵母菌悬液按4:1的体积比充分混匀,紫外线照射处理60 s后稀释涂布,将涂布后的平板置于30 ℃恒温培养箱中,避光培养48 h,待菌落形态稳定后进行计数,并计算氧化损伤酵母细胞的存活率。以VC溶液作阳性对照。氧化损伤酵母细胞存活率根据公式(5)计算:
$ \text{紫外氧化损伤酵母细胞存活率}(\text{%})=\frac{\mathrm{A}_{\mathrm{i}}-\mathrm{A}_{\mathrm{j}}}{\mathrm{A}_{\mathrm{k}}-\mathrm{A}_{\mathrm{j}}}\times100 $
式中,Ak表示未经紫外线照射处理的酵母单菌落数;Ai表示样品组的酵母单菌落数;Aj表示对照组的酵母单菌落数。
参照胡亚平等[27]的方法稍作改动。按1.2.6.1的方法制得酵母菌悬液。取对数期酵母培养液50 mL,4000 r/min离心10 min后收集菌体。用生理盐水洗涤2次至不含培养基颜色后制备菌悬液。按照4:1的比例将不同质量浓度(5、10、15、20、25 mg/mL)的多糖溶液与菌悬液混合后,加入1 mL 2% H2O2溶液,并充分混合,于37 ℃孵育振荡1 h,稀释涂布,在30 ℃条件下培养48 h,并进行菌落计数。以VC为阳性对照,按公式(6)计算酵母细胞存活率:
$ \text{存活率}(\text{%})=\frac{{\mathrm{A}}_{\mathrm{i}}-{\mathrm{A}}_{\mathrm{j}}}{{\mathrm{A}}_{0}-{\mathrm{A}}_{\mathrm{j}}}\times 100 $
式中,Ai表示添加样品后氧化损伤酵母单菌落数;Aj表示氧化损伤酵母单菌落数;A0表示未经处理酵母单菌落数。
实验结果通过每组三个独立实验的均值±标准差(n=3)表示。采用IBM SPSS 20.0进行数据处理分析,组间比较采用tukey检验,显著水平P<0.05。使用OriginPro 2015绘图。
三种平菇多糖的基本组成见表1,其中POP中的总糖含量最高为84.20%±0.28%,显著高于POFP(55.53%±0.09%)和POMP(44.72%±0.17%)(P<0.05);POMP中的蛋白质含量最高为27.00%±0.12%,显著高于POFP(14.50%±0.90%)和POP(10.84%±0.29%)(P<0.05),蛋白质含量是粗多糖的重要检测指标。三种多糖的水分含量由大到小分别为POFP>POP>POMP,水分含量的不同反映出多糖在纯度、活性和提取效率等方面存在的差异。
对三种平菇多糖的相对分子量进行检测,结果显示POFP、POMP、POP多糖均由2种不同分子量的多糖组成,其保留时间接近,结果见表2。三种多糖的分子量的大小范围在375.31~9788.14 Da之间。分子量大小排序为POP>POMP>POFP。这与Hao等[28]对红缘拟层孔菌子实体多糖、菌丝体多糖和发酵液多糖的分子量研究结果类似。有研究表明,分子质量的大小可能会影响多糖的构象、与自由基的相互作用方式及其在生物体系中的稳定性,从而对多糖的抗氧化活性产生显著影响[29]。低分子量多糖因在溶液中具有更好的扩散性,能更易与自由基发生反应,而高分子量多糖由于分子间存在较强的相互作用,更有利于金属离子的螯合[30]。分子量的差异为后续多糖抗氧化活性对比研究提供了重要参考。
DPPH自由基在517 nm处有最强吸收峰,抗氧化剂与之结合会使溶液颜色变浅,吸光度降低,因此抗氧化剂的还原能力与吸光度呈现反比的关系,吸光度越低,表明还原能力越强[26]。三种平菇多糖对DPPH自由基的清除能力如图1所示,其清除能力呈现剂量依赖性,在多糖浓度为1~5 mg/mL范围内,三种粗多糖的DPPH自由基清除能力与多糖溶液的浓度具有正相关关系。将DPPH自由基清除率与多糖浓度进行线性拟合得到回归方程,根据回归方程求出清除率为50%时的多糖浓度即为半清除率浓度IC50,POFP、POMP和POP的IC50分别是3.37、0.82、0.09 mg/mL。当多糖浓度为5 mg/mL时,POP和POMP的清除率都达到90%以上,与VC清除效果相当,并且显著高于POFP的清除率(75.13%±1.97%)(P<0.05),清除率大小排序为POP>POMP>POFP。
ABTS+自由基的检测波长约为734 nm,数值大小与被测样品抗氧化能力有关,数值越小,说明自由基越少,即样品对ABTS+自由基的清除能力越强[20]。三种多糖对ABTS+自由基的清除结果如图2所示。在1~5 mg/mL浓度范围内,三种多糖对ABTS+自由基清除率呈现剂量依赖性。线性拟合得到,POFP、POMP和POP的IC50分别是8.58、6.06、5.09 mg/mL。当多糖浓度为5 mg/mL时,对三种多糖及VC的自由基清除率进行测定,结果发现其清除率由大到小依次为 VC>POP>POMP>POFP。具体来看,在ABTS+自由基清除方面,POP的清除率为49.71%±2.75%,POMP的清除率为40.24%±2.53%,二者均显著高于POFP的清除率(29.67%±2.41%)(P<0.05)。尽管三种多糖对ABTS+自由基的清除率均低于其对DPPH自由基的清除率,但总体趋势保持一致。
OH自由基与活细胞中的任何分子发生反应均会造成损伤,其反应速度非常快,引起的自由基链反应对生物体造成较大范围的损害[28]。不同浓度的三种粗多糖对OH自由基的清除能力如图3所示。在1~5 mg/mL浓度范围内,三类粗多糖对OH自由基清除能力随着浓度增加而增加。将OH自由基清除率与多糖浓度进行线性拟合,POFP、POMP和POP的IC50值分别是0.46、0.91、0.48 mg/mL。当多糖浓度为5 mg/mL时,POP对OH自由基清除率最高为97.25%±1.71%,与VC对OH自由基的清除效果接近,其次是POMP为93.52%±1.61%,两者均显著高于POFP对OH自由基清除率(86.32%±3.01%)(P<0.05)。
抗氧化剂通过自身还原作用给出电子而清除自由基,还原力越强,抗氧化活性越强,其在700 nm处有最大吸收峰,其吸光度与还原力表现出正相关关系[31]。三种粗多糖的总还原力如图4所示。在1~5 mg/mL浓度范围内,三种多糖的总还原力呈现一定的剂量依赖性。当多糖含量为5 mg/mL时,POFP、POMP和POP的总还原力吸光度分别为0.51±0.04、0.38±0.04、0.31±0.03,由大到小排序为POFP>POMP>POP。由此可知,液态发酵平菇多糖总还原力高于菌丝体多糖和子实体多糖。
紫外线照射皮肤容易引起活性自由基和活性氧,其与皮肤衰老和癌症的发生密切相关,抗氧化剂的存在可减少紫外线照射对细胞造成的损伤[32]。如图5所示,三种粗多糖对紫外线损伤酵母细胞均具有一定程度的保护作用,且在试验浓度范围内其保护作用与多糖浓度呈正相关。将酵母细胞存活率与多糖浓度进行线性拟合,POFP、POMP和POP的IC50值分别是23.51、12.34、10.09 mg/mL。当多糖浓度为25 mg/mL时,三种多糖均显著提高了酵母细胞的存活能力(P<0.05);其中,POP对紫外线氧化损伤酵母细胞的保护效果尤为突出,其存活率可达96.05%±4.02%,显著大于POMP(72.37%±1.99%)和POFP(53.29%±2.63%)(P<0.05),甚至超过了阳性对照样VC对损伤酵母细胞的保护效果,这表明平菇多糖对氧化损伤酵母细胞具有显著的保护作用,未来其有可能成为防晒霜和美容护肤品开发的重要活性原料。
H2O2常被用于诱导细胞或组织发生氧化应激损伤,通过产生OH自由基,破坏机体内的活性分子,从而发生病变[33]。如图6所示,三种粗多糖对H2O2氧化损伤酵母细胞均具有保护作用,且均在15 mg/mL时对酵母细胞的保护作用达到最大值并趋于平稳。将酵母细胞存活率与多糖浓度进行线性拟合,POFP、POMP和POP的IC50值分别是9.35、8.57、7.79 mg/mL。当多糖浓度为15 mg/mL时,POP、POMP和POFP多糖组H2O2氧化损伤酵母细胞存活率分别为91.60%±3.26%、84.20%±3.06%和74.80%±2.88%;而且,POP多糖组H2O2氧化损伤酵母细胞存活率与VC组相当,进一步说明平菇多糖具有显著的抗氧化活性。
本文对比了平菇发酵液多糖(POFP)、平菇菌丝体多糖(POMP)和平菇子实体多糖(POP)的基本组成、分子量及其抗氧化活性。结果表明:三种多糖的基本组成存在显著差异,其中POP中的总糖含量最高(84.20%±0.28%),显著高于POFP(55.53%±0.09%)和POMP(44.72%±0.17%);POMP的蛋白质含量(27.00%±0.12%)显著高于POFP(14.50%±0.90%)和POP(10.84%±0.29%)。三种多糖均由两种不同分子量的多糖组成,其分子量由大到小顺序为:POP>POMP>POFP。体外抗氧化活性结果显示,在一定的浓度下,三种多糖均有显著的DPPH自由基、ABTS+自由基、OH自由基清除能力,并能显著提高氧化损伤酵母细胞的保护效果。其中5 mg/mL的POFP的总还原力为0.51±0.04,高于POMP和POP。POP保护作用最强,25 mg/mL和15 mg/mL的POP可将紫外线和H2O2氧化损伤酵母细胞活力分别恢复至96.05%±4.02%和91.60%±3.26%,与VC处理组效果相当。本研究不仅采用液态发酵技术高效制备平菇多糖,为抗氧化资源库提供了新的选择,也为平菇多糖在功能性食品和美容护肤品等领域的应用提供了技术支撑。未来可进一步探究平菇多糖基本组成、分子量和结构与其抗氧化活性之间的关系,为基于平菇多糖的抗氧化产品开发提供参考。

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2026年第47卷第5期
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doi: 10.13386/j.issn1002-0306.2025050071
  • 接收时间:2025-05-12
  • 首发时间:2026-10-24
  • 出版时间:2026-03-01
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    1天津科技大学,工业发酵微生物教育部重点实验室,天津 300457
    2军事科学院系统工程研究院,北京 100010

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韩培培(1983−),女,博士,教授,研究方向:生物工程,E-mail:。
郝利民(1969−),男,博士,教授,研究方向:特殊功能食品与食品生物技术,E-mail:。
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2种不同金属材料的力学参数
科
Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)
属
Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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