植物精油对微生物作用机制及在鱼类保鲜中的应用研究进展

蓝蔚青1,2,郎艾1,梅俊1,2,谢晶1,2*

1(上海海洋大学 食品学院,上海 201306) 2(上海水产品加工及贮藏工程技术研究中心,食品科学与工程国家级实验教学示范中心(上海海洋大学),上海 201306)

摘 要 鱼类的腐败变质一直以来都是水产品安全问题的关键,也是困扰水产行业发展的难点。植物精油是大量芳香植物的次生代谢物质,具有抗菌和抗氧化作用,是一种天然生物保鲜剂。该文在分析鱼类水产品腐败变质原因与植物精油对微生物作用机制的基础上,综述了植物精油在鱼类保鲜中的应用研究进展,提出了存在问题与解决办法,展望了植物精油在鱼类保鲜中的发展趋势,以期为后期拓展其应用领域提供理论参考。

关键词 植物精油;作用机制;鱼类产品;保鲜

我国海域辽阔,拥有丰富的海洋生物资源,发展潜力巨大。据《中国渔业统计年鉴2020》统计显示,我国2019年的水产品总产量为6480.36万t,比2018年增长0.35%[1]。鱼类水产品中的蛋白质、不饱和脂肪酸与水分含量高,是人类营养物质的重要来源[2]。然而,由于其具有易腐性,流通期间会受到保鲜技术与贮藏条件制约,在微生物和酶的作用下发生腐败,产生不良口感、色泽与风味,甚至有害物质,降低了其营养与经济价值[3]。目前,我国鲜活鱼类水产品捕后腐损率达15%,而发达国家则低于5%[4],这已成为制约我国水产业发展,影响养殖户与渔民增收,提高市场竞争力的重要因素。为降低水产品流通期间的损失,开展鱼类保鲜技术研究意义重大。

现有研究表明,鱼类水产品贮藏后期的腐败变质主要由微生物活动引起[3,5]。新鲜鱼类中的微生物种类繁多,主要为革兰氏阴性杆菌[6]。其中少数微生物能适应贮藏环境,快速生长繁殖成为优势菌群,这些微生物称为特定腐败菌(specific spoilage organisms,SSO)[7]。SSO的种类受鱼种类、贮藏温度与环境条件等因素影响[8]。如未经冷藏鲜鱼的SSO是弧菌属等革兰氏阴性发酵型细菌,冰鲜淡水鱼和海水鱼的SSO是假单胞菌与希瓦氏菌,CO2气调包装中鱼类水产品的SSO是明亮发光杆菌与乳酸菌[9]。微生物主要通过代谢产生的蛋白酶、酯酶等,其能分解鱼类产品中的蛋白质、脂质等营养物质,使鱼的肌肉质构特性与营养改变,导致腐败[10-12];同时,微生物还会形成被膜等物质,使鱼发黏[13],并进一步代谢氨基酸与寡肽,产生H2S、三甲胺、腐胺与尸胺等小分子挥发性风味物质,产生异味,进而使其品质劣变[10]

鱼类常用的保鲜方式有物理保鲜、化学保鲜与生物保鲜[3]。生物保鲜法是利用从生物中提取或用微生物工程技术获得的天然、对人体安全的保鲜剂来进行保鲜的方法[14-15]。此法由于符合消费者对天然绿色食品的追求,因此受到广泛关注。生物保鲜剂按其来源不同,可分为植物源、动物源、微生物源和酶类4种。动物源生物保鲜剂主要有壳聚糖、蜂胶与鱼精蛋白等,其溶解性与抑菌性易受环境影响,且应用领域有限;微生物源生物保鲜剂主要有乳酸链球菌素(Nisin)、抗菌肽与聚赖氨酸等,环境条件会对其抑菌效果产生影响;酶类生物保鲜剂主要有溶菌酶与脂肪酶等,其价格昂贵,且在鱼类保鲜中的应用还不深入;植物精油属植物源生物保鲜剂,为芳香植物的高度浓缩脂溶性天然化合物,具有挥发性、高渗透与味道浓郁等特点[14],与其他类型的生物保鲜剂相比,植物精油具有抗菌与抗氧化能力,且用量少、经济适用。植物精油中的化学成分复杂多样,主要含萜烯类、醇类与酮类等多种成分,每种精油均由2~3种主要成分组成,占总量的70%,其主要成分决定了不同精油的性质[16]。由于植物精油含有丰富的化学成分,因此具有杀虫、抗氧化和抗菌等多种生物学活性[13]。多项研究证实,植物精油对食源性致病菌和腐败菌有抑制作用,可达到防腐保鲜效果[17]

研究表明,植物精油的抗菌、抗氧化活性源于芳香化合物中的萜烯化合物、酸、醇、酯、醛、酮、胺类、环氧化物与硫化物等成分[18-19],鱼类保鲜常用植物精油、主要成分与作用菌株如表1所示。

表1 鱼类保鲜常用植物精油种类、主要成分与作用菌株

Table 1 Species,main components and functional strains of plant essential oils commonly used in fish preservation

精油名称主要成分主要作用菌株参考文献肉桂精油肉桂醛希瓦氏菌、气单胞菌、假单胞菌、大肠杆菌、金黄色葡萄球菌、芽孢杆菌[20-21]丁香精油丁香酚希瓦氏菌、气单胞菌、假单胞菌、大肠杆菌、金黄色葡萄球菌、芽孢杆菌[20]百里香精油百里酚金黄色葡萄球菌、枯草芽孢杆菌、大肠杆菌、腐败希瓦氏菌、副溶血性弧菌[20-22]桉叶精油1,8-桉叶素大肠杆菌、金黄色葡萄球菌、腐败希瓦氏菌、副溶血性弧菌[23]牛至精油麝香草酚1-甲基-2-异丙基-苯荧光假单胞菌、大肠杆菌、腐生葡萄球菌、副溶血性弧菌、腐败希瓦氏菌[6][21][24]迷迭香精油α-蒎烯、1,8-桉叶素大肠杆菌、枯草芽孢杆菌[25]

植物精油中的香芹酚可改变菌体细胞膜通透性,使钾离子泄露,干扰质子运动,抑制ATP合成[26];植物精油的异硫氰酸类硫化物会诱导菌体细胞周期阻滞,导致细菌丝状化,影响细胞流动性和稳定压力,达到其抑菌目的[27];植物精油还可抗脂质氧化,延缓其腐败[28]。由于植物精油的化学成分复杂,其抑菌作用可能是一种或多种成分协同作用的结果[18]。植物精油对微生物的作用靶点及抑菌机制如图1所示。

图1 植物精油对微生物的作用靶点及抑菌机制[29]

Fig.1 Target and antibacterial mechanism of plant essential oil on microorganisms[29]

注:QS.群体感应(quorum sensing);FtZ.丝状温度敏感蛋白(filamentous-sensitive protein Z)

1 植物精油在鱼类保鲜加工中的应用

植物精油主要采用熏蒸、浸渍或喷洒等方式,通过抑菌与抗氧化等发挥作用,应用于鱼类保鲜[20]

1.1 单一植物精油保鲜

微生物是引起鱼类水产品腐败变质的主因,微生物数与其新鲜程度关系密切。新鲜鱼类的细菌总数为102~104 CFU/g,达到初期腐败时为106 CFU/g,当增至107~108 CFU/g时,便能感受到强烈腐臭味[30]。植物精油中含有多种有效成分,具有良好的抑菌活性。杨胜平等[6]用体积分数0.062 5%牛至精油处理接种荧光假单胞菌的冷藏三文鱼,发现贮藏12 d时,其菌落总数对数值较对照组样品低1.34 lg CFU/g,总挥发性盐基氮(total volatile basic nitrogen,TVB-N)降低7.48 mgN/100 g,腐胺与尸胺含量分别降低48.5%与33.5%;雷志方等[31]用质量分数2%姜精油处理金枪鱼后于4 ℃条件下贮藏,得出对照组样品在第3 天时K值达47.2%,而姜精油处理组样品到第10 天时,K值仍未超40%,处理组样品到第10 天时的组胺含量为62.43 mg/kg,对照组已达82.33 mg/kg;常开颜等[32]用90 mg/kg肉桂醛处理鱿鱼鱼丸后于4 ℃条件下贮藏,发现肉桂醛可有效抑制鱼丸中的微生物生长,延缓TVB-N值与硫代巴比妥酸值上升、减缓pH值与硬度值变化,提高鱼丸的咀嚼度与感官品质,能延长货架期6~7 d。

1.2 植物精油复配保鲜

绝大多数植物精油都有抑菌作用,但不同种类的植物精油杀灭微生物的类型还有差异。因此,将不同种类的植物精油复配,能发挥其各自抑菌效能,提升综合效果。其中,孟玉霞[20]用4 μL/mL山苍子精油与6 μL/mL肉桂醛浸泡处理大黄鱼后,4 ℃冷藏12 d,植物精油复配组样品的微生物数均显著低于对照组(P<0.05);侯春宇[33]探究了百里香酚与丁香酚、松油醇、肉桂醛复配处理对大黄鱼鱼片冷藏期间品质变化影响,得出体积分数4.785%百里香酚与体积分数7.81%丁香酚联合抗菌效果最强,其微生物、氨基酸与感官指标均优于其余各组,能明显减缓大黄鱼贮藏期间的品质劣变。

1.3 与其他生物保鲜剂复配保鲜

近年来,也有相关科研人员开展了植物精油与其他生物保鲜剂复配用于鱼类保鲜。余小亮[21]将茶多酚与肉桂精油按1∶1体积比例复配后处理鲈鱼后冷藏,复合保鲜剂有效抑制了腐败微生物繁殖与鱼体内源酶活性,延缓营养物质降解,以3 g/L复配处理组效果最佳;赵厚菲[34]研究了2 g/L肉桂醛与3 g/L溶菌酶复合涂膜对美国红鱼冷藏货架期的影响,证明复合涂膜综合效果优于单一涂膜,能使其货架期延长5~10 d;REZAEIFAR等[35]研究了体积分数2%壳聚糖与0.5%柠檬马鞭草精油、1%柠檬马鞭草萃取物复合涂膜对真空包装虹鳟鱼品质变化的影响,复合处理能减缓样品冷藏期间各项指标的升高,使其冷藏货架期延至16 d;RAHIMABADI等[36]采用体积分数0.2%野蔷薇精油与150 000 IU/kg Nisin结合处理虹鳟鱼,与对照组样品相比,精油复配组能延长虹鳟鱼冷藏货架期3 d;SHAHINFAR等[37]采用微胶囊化体积分数0.5%紫草精油与200 IU/kg Nisin结合可改善冷藏鱼汉堡的感官品质。

1.4 与抗菌包装组合保鲜

多数植物精油具有厚重气味、易挥发、高渗透的特点,故直接添加到鱼类水产品中会有感官不可接受、精油发挥作用时间短与保鲜效果不佳等问题。部分学者用精油与包装材料组合的抗菌活性包装来改善产品感官特性,达到保鲜目的[38]。目前植物精油在鱼类抗菌包装体系中,可制成膜液涂于鱼类水产品表面或食用性膜包裹在产品表面。刘善智[39]将2 mg/mL壳聚糖+1.5 mg/mL三聚磷酸钠+2 mg/mL迷迭香精油,按5∶2∶1的体积比例复配,在pH值为4.0的条件下制备成迷迭香精油壳聚糖纳米粒,涂抹在草鱼肉上后予以冷藏,该处理能有效保护鱼肉色泽,改善冷藏期间其感官品质,使其冷藏货架期达7 d;FARSANIPOUR等[40]将龙蒿精油与分离乳清蛋白、壳聚糖制成可食涂层作用于康氏似鲹,得出精油与分离乳清蛋白、壳聚糖涂层组样品的冷藏货架期为10 d,较对照组延长了6 d,可见其具有较强的保鲜效果;杨辉[41]发现添加质量分数1%丁香精油与质量分数1%葡萄籽精油的乙烯-乙烯醇共聚物薄膜能使草鱼冷藏货架期达8 d;ATREA等[42]在添加体积分数0.4%牛至精油后真空包装地中海章鱼,可使其冷藏货架期延长20 d。

1.5 其他方式协同处理保鲜

虽然植物精油抑菌效果好,但鱼类水产品腐败变质的机理复杂,受外界条件影响大,单一植物精油作用效果有限。因此,还可将植物精油处理结合其他保鲜方式进行组合保鲜。目前,以植物精油为基础,辅以气调[43-44]、被膜、超高压处理及添加其他抗菌物质等作用于鱼类水产品,得到较好效果。其中,刘蒙佳等[45]将蒙脱土与体积分数2%丁香精油、壳聚糖制成复合抑菌保鲜衬垫,协同55%O2+30%CO2+15%N2气调包装贮藏金鲳鱼,结果得出复合抑菌垫协同气调处理在-1 ℃下贮藏,可延长金鲳鱼货架期约4 d;YILDIZ等[46]用体积分数1%百里香精油结合50%CO2+50%N2气调包装处理热熏蒸后的虹鳟鱼,精油与气调包装协同作用可有效抑制微生物的生长,使产品冷藏货架期延长40 d。将植物精油加入渗透液中,利用食物与周围溶液的渗透压差,降低水分活度,抑制微生物生长,这种双重杀菌作用可延长冷冻渗透预处理鱼制品的货架期,提升其商业价值;TSIRONI等[47]用500 g/L高葡萄糖当量麦芽糊精与体积分数0.5%香芹酚对金枪鱼片进行渗透处理,在5 ℃贮藏条件下,可延长货架期8 d。

2 存在问题与解决方法

2.1 安全性

大部分植物精油从古至今都在使用,安全性高,是一般公认安全(Generally Recognized as Safe,GRAS)产品[48]。另外,植物精油在食品工业中使用时,其化学与毒理学特性表征已得到指数评估[48]。自2018年4月1日起,肉桂、香樟、小花百里香、迷迭香等156种芳香植物精油被《联邦法规》认可在监管限制下安全。因此,根据“安全使用的悠久历史”和“限制原则”等概念判断植物精油是在预期使用条件下是安全的。但有人提出,由于植物精油提取自植物,而植物有可能含有农药残留,导致植物精油被污染,将其用于鱼类保鲜,会影响其安全性[49]。目前这一说法尚需后续验证。

2.2 对鱼类水产品风味和色泽产生影响

植物精油具有刺激性气味,作用于鱼类水产品会对其风味、色泽等感官可接受性带来影响。如将2 μL/mL肉桂醛溶液浸渍大黄鱼与大菱鲆15 min,使其气味与色泽接受度降低[20];CAI等[50]研究发现,4 μL/L丁香、薄荷与孜然精油熏蒸红鼓鱼片后,鱼片的气味感官良好,但体积分数为8 μL/L时,其气味不可接受;TSIRONI等[47]用0.5%香芹酚渗透液处理金枪鱼鱼片后,有轻微异味。因此,将精油做成活性包装[39]、加入可食用包装膜[40]、制备成纳米乳剂[51]用于食品中,不仅可降低精油对风味影响,避免精油直接涂抹导致色泽不佳,且可使精油及其抑菌成分缓控释放,提高保鲜效果。其中,CHUESIANG等[52]用4 g肉桂精油制成纳米乳液处理25 g亚洲鲈鱼鱼片,得出经生物聚合物纳米载体包裏的肉桂精油不仅在抑制细菌生长、减缓不良化学反应方面表现出更好效果,且防止了样品色泽与质地改变;VITAL等[53]研究了体积分数0.1%生姜和0.1%牛至精油褐藻酸盐涂层鱼片的质量和感官接受度,发现精油涂层处理组样品的脂质氧化、颜色与质量损失相应降低。

2.3 作用效果易受影响

植物精油的作用效果会受种类、食品基质成分与物理结构等因素影响,同种精油对不同腐败菌的抑制效果存在差异。杨辉[41]用体积分数2%丁香精油处理草鱼可有效减缓菌落总数的升高,而贾亦森[54]用30 μL/L丁香精油处理大鲵分割肉发现其抑菌效果不显著。已有研究发现,植物精油的功效可在与某些食品基质(如蛋白质、脂肪、碳水化合物)与食品pH值相互作用的同时进行改良。如在酸性食品中,由于低pH值使植物精油的疏水能力更强,其在细菌膜中溶解的能力随之增强,细菌对精油的敏感性增加[55]。孟玉霞[20]实验得出,2倍最小抑菌浓度的山苍子精油对希瓦氏菌的作用效果仅在第3 天和12 d时减少了1.3~1.4个对数值,其他时间点无显著差异。此外,温度、气体组成、包装方法、微生物性质与初始浓度等也会对植物精油的作用效果产生影响[55-56]。因此,要使植物精油稳定高效发挥其抑菌作用,应根据不同精油的特性,选择合适的作用方式和环境,还可将精油中有效抑菌成分优化后制成保鲜剂,增强其作用效果。

2.4 作用机制尚未完全明确

植物精油具有广谱抗菌性,但其抗菌化学成分复杂、可变性高,与细菌作用的靶点多,可能不是单一机制形成抑菌作用。目前研究认为,植物精油主要作用于细菌的细胞膜,同时影响细菌代谢与核酸物质,达到其抑菌效果。

植物精油中高浓度醛、酚类物质通过破坏细胞膜、细胞质膜,影响细菌正常生命活动,MYSZKA等[57]发现120 nL/mL青椒精油可改变铜绿假单胞菌菌体膜流动性,抑制弹性蛋白酶活性,降低绿脓杆菌素的生物合成,减缓鱼制品中菌体生长和毒力;陈梦玲等[24]发现腐生葡萄球菌经体积分数0.25%牛至精油处理后,其细胞壁、细胞膜通透性与完整性遭到破坏,菌体内的活性物质可与精油的疏水活性物质相互结合,影响菌体生长与代谢;SHARMA等[58]认为革兰氏阴性菌的外膜具有强大的脂多糖分子,使各种抗菌剂不可渗透,但PATRA等[59]研究结果证实了植物精油作用能使菌体内的活性氧过量积累,导致胞内DNA单链断裂损伤,对革兰氏阴性菌具有较好抗菌作用。还有研究发现,胡椒精油能诱导酿酒酵母产生活性氧[60]。因此,植物精油的作用机制还有待于后期深入研究。

3 前景与展望

采用天然保鲜剂替代化学防腐保鲜剂是食品行业和消费市场发展的大趋势,植物精油用于果蔬保鲜取得了良好效果。然而,由于其成分复杂、作用靶点多样、抑菌抗氧化机制尚未完全明确、气味影响产品感官、某些精油或组分对人体的安全性还需系统评价等特点,其在鱼类保鲜中的应用仍存在一些问题。因此,在探究某类精油能否用于鱼类保鲜时,可研究精油的抑菌抗氧化主要化学成分、抑菌机制及作用条件,并结合其他保鲜技术和鱼类水产品特点优化保鲜效果,达到经济安全适用的目的。在GB 2760—2014《食品安全国家标准 食品添加剂使用标准》中,仅肉桂醛(CNS17.012)可用作防腐剂,还限制了使用范围,基本所有精油被归类于食用香料物质,这限制了精油在鱼类防腐保鲜中的应用。但总体来讲,植物精油因其安全性高,抑菌力强,且历年来许多实验证明其安全性与优良的保鲜效果,将植物精油予以精制提取,或与其他生物保鲜剂、抗菌包装、保鲜技术等复合处理,能进一步降低其使用量,提升其综合作用效果,是一类前景较好的生物保鲜剂。

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Research progress on the action mechanism of plant essential oils on microorganisms and its application in the preservation of fish products

LAN Weiqing1,2,LANG Ai1,MEI Jun1,2,XIE Jing1,2*

1(College of Food Science and Technology,Shanghai Ocean University,Shanghai 201306,China) 2(Shanghai Aquatic Products Processing and Storage Engineering Technology Research Center,National Experimental Teaching Demonstration Center for Food Science and Engineering(Shanghai Ocean University),Shanghai 201306,China)

ABSTRACT The deterioration of fish products has always been the key problems to food safety,but also the developmental difficulty of fish products industry.Plant essential oils are the secondary metabolite of a large number of aromatic plants.It has antibacterial and antioxidant functions,which is a natural biological preservative.Based on the analysis of the spoilage causes of fish products and the action mechanism of plant essential oils on microorganisms,the application of plant essential oils in the preservation of fish products is reviewed,the existing problems and solutions are proposed,the future development trend of plant essential oils in fish products industry is also prospected so as to provide the theoretical references for expanding its application in the future.

Key words plant essential oil;action mechanism;fish products;preservation

DOI:10.13995/j.cnki.11-1802/ts.025567

引用格式:蓝蔚青,郎艾,梅俊,等.植物精油对微生物作用机制及在鱼类保鲜中的应用研究进展[J].食品与发酵工业,2021,47(13):296-302.LAN Weiqing,LANG Ai,MEI Jun,et al.Research progress on the action mechanism of plant essential oils on microorganisms and its application in the preservation of fish products[J].Food and Fermentation Industries,2021,47(13):296-302.

第一作者:博士,高级工程师(谢晶教授为通讯作者,E-mail: jxie@shou.edu.cn)

基金项目:国家自然科学基金面上项目(31972142);现代农业产业技术体系建设专项(CARS-47-G26);上海水产品加工及贮藏工程技术研究中心能力提升项目(19DZ2284000)

收稿日期:2020-08-20,改回日期:2020-10-26