白鲢鱼脱腥及其低盐鱼糜制备的研究
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摘要
白鲢鱼抗病、生长快、成本低,野生和养殖量都很高。把白鲢鱼加工成高附加值的鱼糜,是有效利用白鲢鱼资源的途径之一,可以显著地提高白鲢鱼的附加值,提高其经济效益。但是,腥味重和凝胶强度低限制了白鲢鱼鱼糜的产业发展。
     本论文主要目的是制备高品质脱腥白鲢鱼低盐鱼糜,研究了白鲢鱼腥味检测方法、腥味产生机理、脱腥方法以及利用微波加热提高低盐鱼糜凝胶强度的工艺和机理。
     采用感官评定、固相微萃取-气相-质谱(SPME-GC-MS)、同时蒸馏萃取-气相-质谱(SDE-GC-MS)结合时间-强度法(OSME)和香气稀释分析法(AEDA)两种嗅闻方法等技术检测和鉴定了白鲢鱼肉的挥发性风味物质。E,E-2,4-庚-二烯醛(E,E-2,4-heptadienal)、E,E-2,4-癸-二烯醛(E,E-2,4-decadienal)、E-2-癸烯醛(2-decenal)是白鲢鱼鱼腥味的特征物质。己醛(hexanal)和1-辛烯-3-醇(1-octen-3-ol)分别是引起白鲢鱼青草味和蘑菇味的特征物质。
     采用微波辅助蒸馏-固相微萃取-气相-质谱联用技术(MD-SPME-GC-MS)检测了白鲢鱼肉中土霉味物质土味素(GEO)和2-甲基异莰醇(MIB)。微波辅助蒸馏提取白鲢鱼肉中的MIB和GEO最佳条件是微波功率350W/10g鱼肉,微波蒸馏时间8 min。
     评价了电子鼻对白鲢鱼腥味的检测效果。电子鼻对鱼腥味、哈喇味、土霉味都能做出特异响应,且电子鼻检测哈喇味、鱼腥味、土霉味的结果与感官评定结果相关系数(R2)分别高达0.9857、0.9207和0.9909;所以电子鼻能准确检测白鲢鱼的腥味。
     研究了白鲢鱼肌肉脂肪氧化和鱼腥味产生的机理。反相高效液相(RP-HPLC)的结果表明白鲢鱼肉中的脂肪氧合酶(lipoxygenase, LOX)主要是12-LOX。在鱼肉模拟体系中,LOX引发鱼肉脂肪氧化速度最快,最先形成脂肪酸过氧化物;而血红蛋白、Fe~(3+)在脂肪氧化的初始阶段裂解脂肪酸过氧化物的活力很高。SPME-GC-MS-O和感官评定的结果表明,白鲢鱼鱼腥味主要是由12-LOX催化鱼肉脂肪中的亚麻酸生成E,E-2,4-庚-二烯醛(E,E-2,4-heptadienal)引起的。
     比较了水洗法、酸法、碱法、酵母细胞液处理法对白鲢鱼肉的脱腥效果。碱法脱腥工艺的蛋白回收率较高,鱼腥味、土霉味脱除效果均较好,土霉味物质浓度降低到0.5μg/kg以下。葡萄酒酵母细胞液脱鱼腥味效果最好,葡萄酒酵母细胞液脱腥的适宜条件为25 oC、pH 6.5、2 h。
     应用顶空-气相色谱法(HP-GC)测定了鱼糜蛋白对腥味物质的吸附率、亲和常数(K)、极限吸附量(n),研究了酸碱法的脱腥机理。碱法脱腥降低了鱼糜蛋白对腥味物质亲和常数(K),腥味物质易于从鱼糜蛋白质分子上解吸,鱼糜蛋白对腥味物质的吸附率降低,所以脱腥效果较好。而酵母细胞液脱腥的机理是醇脱氢酶、醛脱氢酶把鱼肉中有腥味的醛转化成无腥味的醇和酸。
     采用差示扫描量热仪(DSC)、十二烷基磺酸钠-聚丙烯酰氨凝胶电泳(SDS-PAGE)、扫描电子显微镜(SEM)等研究了酸碱法脱腥对白鲢鱼蛋白质分子结构的影响。酸法脱腥使肌球蛋白完全变性,肌动蛋白含量降低;碱法脱腥鱼糜的肌球蛋白基本没有变性,肌动蛋白含量最高。水洗脱腥鱼糜凝胶具有三维网状结构,弹性较好;酸法脱腥鱼糜凝胶没有形成规则的网状结构,因此其凝胶强度最低;碱法脱腥鱼糜凝胶的网状结构不规则,大多数蛋白聚集成颗粒状,堆积致密,因而其凝胶硬度最高。白鲢鱼肌原纤维蛋白在酸碱处理过程中发生明显降解和变性聚集,这是造成酸、碱脱腥鱼糜凝胶性质变差的主要原因。特别是酸处理使肌球蛋白完全变性,分子表面疏水性增强,溶解性变差,在热致胶凝过程中聚集速度太快,蛋白质分子间不能充分相互作用以形成良好的三维网状结构,所以凝胶强度最低。
     采用微波加热制备低盐鱼糜凝胶。白鲢鱼碱法脱腥低盐鱼糜微波加热胶凝的合适工艺为:添加大豆分离蛋白(SPI)3% (w/w),在胶凝起始温度40 oC下保温1 h,微波功率中档(15 W/g)加热60 s,所得鱼糜凝胶强度达2077.4±25.5 g×cm。微波加热胶凝能有效避免肌球蛋白降解,鱼糜蛋白通过S-S键、P-N键、羰-氨键等交联;而且超微结构的结果表明微波加热胶凝使鲢鱼碱法脱腥鱼糜蛋白质聚集体充分展开和相互作用,形成三维网状结构,所以微波加热胶凝能显著提高低盐鱼糜凝胶的强度(P<0.05)。
     提出了低盐鱼糜微波加热胶凝机理:第一阶段,较大蛋白聚集体(直径1-2μm)在微波高密度加热时分开成细丝状蛋白聚集体(直径0.1-0.2μm),它们互相交叉缠绕,形成非常细密的网状结构;第二阶段,邻近的细丝状蛋白重新聚集成使网孔孔壁变厚(1-2μm),形成三维网状结构的凝胶。
Silver carp (Hypophthalmichthys molitrix) is widely raised in China, due to its quick growth and resistance to stress, diseases and rough handling. In order to improve the added-value of silver carp, it is economic to utilize the silver carp as raw material of surimi. However, the development of silver carp surimi is limited due to its fishy off-flavor and low gel strength.
     In the present thesis, the silver carp is evaluated to prepare into fishy-free, low salt high quality surimi. The main content of the present study includes the detection of off-flavor of silver carp mince, the mechanism of fishy odour formation, the methods of deodorization, and improving the gel strength of the low salt gel.
     The off-flavors of silver carp mince were detected using sensory evaluation, solid micro-extraction- gas chromatography- mass chromatography (SPME-GC-MS), simultaneous distillation extraction- gas chromatography- mass chromatography (SDE-GC-MS), and electronic nose. The fishy odour compounds of silver carp mince were identified as E, E-2, 4-heptadienal, E, E-2, 4-decadienal, and E-2-decenal. The microwave assistant distillation-SPME-GC-MS is suitable for detection of earthy-muddy off-flavor compounds of geosmin and 2-methylisoborneol of silver carp mince, the optimized conditions for microwave assistant distillation were: microwave power output 350 W and distillation time 8 min.
     The electronic nose could detect the fishy, rancid, earthy-muddy odour. And the results of electronic nose for rancid odour and earthy-muddy odour were consistent with the results of sensory evaluation, the R2 between them were higher than 0.98.
     The mechanism of the lipid oxidation and fishy odour formation was studied. The main lipoxygenase (LOX) in silver carp muscle was identified as 12-LOX. Among the LOX, Fe~(3+), and hemoglobin, the LOX caused the fastest lipid oxidation, while the latter two catalyzes mainly decompose the peroxides during the initial phase of lipid oxidation. The fishy odour of silver carp mince was mainly caused by the E, E-2, 4-heptadienal, which stem from the oxidation of linolenic acid catalyzed by 12-LOX.
     The effects of water-washing, acid treatment, alkali treatment, and yeast cytosol treatment on the removal of off-flavor of silver carp mince were compared. Alkali treatment deodorizing process obtained a high recovery yield of protein, good effect of the removing of fishy and earthy-muddy odour, the retained earthy-muddy odour compounds were less than 0.5 ug/kg. The cytosol of yeast could reduce the TBARS of silver carp mince significantly (p<0.05), and the grape wine yeast obtained the best result. The optimized conditions for deodorization induced by grape wine yeast cytosol were 25 oC, pH 6.5, for 2 h. However, their mechanism of deodorizing were different, the alkali process reduced the affinity constant (K) between the off-flavor compounds and surimi, thus, removed the off-flavor. The alcohol and aldehyde dehydrogenase of the yeast cytosol were responsible for the removing of fishy odor of silver carp mince.
     Acid process denatured the myosin fully, and reduced the content of actin; while alkali process remained the nature of myosin and recovered the most actin. Gel of water washed surim forming well-regulated three-dimension network, resulting in good elasticity and high gel strength. Acid deodorized surimi could not form three-dimension network, thus, its gel strength was the lowest. Alkali deodorized surimi formed three-dimension network but bad-regulated, and the protein aggregated as particles, thus, the gel showed the highest hardness.
     The myofibril of silver carp underwent considerable degrading and aggregation during the acid and alkali treatment, that were the main reasons of the deterioration of the gel qualities of acid and alkali deodorized surimi. Especially, after the acid treatment, the myosin was denatured absolutely, the surface hydrophobility increased, the solubility decreased, and aggregated too fast to interact with each other to form well-regulated three dimension network, resulting in the lowest gel strength.
     The optimized conditions for preparing low salt gel from alkali deodorized surimi of silver carp using microwave heating are: adding isolated soy protein 3% (w/w); setting at 40 oC for 1 h, followed by microwave heating at middle power intensity for 60 s, the obtained gel strength was 2077.4±25.5 g×cm. Microwave heating effectively expanded the aggregated proteins, and the expanded proteins interacted with each other adequately, forming well-regulated three dimension network; furthermore, the microwave heating protected the surimi from degrading, and there were more covalent bonds formed during microwave heating; therefore, the microwave heating improved the gel strength of low salt gel significantly (p<0.05).
     The gelling mechanism of low salt gel induced by microwave heating was concluded as: in the first stage, big protein aggregates (diameter: 1-2μm) were expanded because of the high intensity of microwave, forming the filament-like protein aggregates (diameter: 0.1-0.2μm), they enwind with each other and formed very dense, mussy network; in the second stage, the neighboring filament-like protein aggregates recombined via hydrophilic interaction, hydrogen bonds, and S-S bonds, forming well-regulated three dimension networks with thicken wall (diameter: 1-2μm).
引文
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