酶解法提取黑木耳中胶原蛋白的工艺优化

为了提高提取黑木耳中胶原蛋白得率,研究黑木耳中胶原蛋白提取工艺路线优化酶法提取的最适条件,采用单因素试验与多元线性回归试验设计相结合的试验方法,经过数据计算统计分析,由回归方程可知,在一定范围内得率与pH值呈正相关,与液料比呈正相关,与酶添加量呈负相关,与酶解时间呈负相关,得出优化工艺参数为:采用胰蛋白酶,酶解pH值为8.0,液料比为52.5:1mL/g,酶添加量为8%,酶解时间为2h,其中在此条件下从黑木耳中提取胶原蛋白得率达到1.091%。研究结果为植物性胶原蛋白制备提供了参考。     

速溶龙眼粉加工的酶解提取与喷雾干燥工艺优化

为了建立速溶龙眼粉的加工技术,运用均匀设计法优化了龙眼干中可溶性固形物酶解提取工艺条件,采用响应面分析法优化了其提取液的喷雾干燥工艺条件,结果表明：以龙眼果肉干（含水率13.62 %）为原料,用果胶酶和纤维素酶同时酶解提取,pH3.1,酶解温度52℃,酶解时间160 min,果胶酶用量0.6‰,纤维素酶用量0.15‰,龙眼干中可溶性固形物得率达85.26%;当酶解提取液中可溶性固形物浓度达25%时,采用喷雾干燥法,选取麦芽糊精为助干剂,其与可溶性固形物含量比0.8：1,热风温度185℃,热风流量26.47 m3/h,入料流量0.20 L/h,在此条件下,龙眼粉得率为48.58%,含水率＜5%,水溶性良好,色泽风味佳。由此说明,以龙眼果肉干为原料采取酶解提取与喷雾干燥相结合的工艺可有效加工速溶龙眼粉。     

金线鱼鱼皮抗冻蛋白酶法制备工艺优化

为了探讨金线鱼鱼皮抗冻蛋白的酶法高效制备工艺,该文首先探究不同蛋白酶酶解获得的酶解产物的热滞活性(thermal hysteresis activity,THA)以及不同分子区间的酶解产物的热滞活性。随后以抗冻蛋白得率为指标优化了底物与酶的比例、酶解时间、酶解温度以及p H值,并采用响应面法探究抗冻蛋白的最佳酶解工艺。最后使用差示扫描量热仪(differential scanning calorimeter,DSC)测定验证抗冻蛋白的热滞活性。结果表明:在碱性蛋白酶、中性蛋白酶、木瓜蛋白酶以及复合蛋白酶4种酶的最佳酶解条件下,碱性蛋白酶酶解所获得抗冻蛋白热滞活性最大,选择碱性蛋白酶作为制备金线鱼鱼皮抗冻蛋白的最佳蛋白酶;截留分子量为≥5~10 k Da鱼皮蛋白的抗冻效果最佳,热滞活性达1.20?0.07℃,确定≥5~10 k Da鱼皮蛋白得率作为酶解制备工艺的指标;得到金线鱼鱼皮抗冻蛋白最佳的酶解制备工艺条件为:底物与酶比例为32∶1、酶解时间为147 min、酶解温度为50℃和p H值为9,在此条件下,金线鱼鱼皮抗冻蛋白得率为49.25%±1.34%。相比于牛血清蛋白(bovine serum albumin,BSA),其降低了溶液起始结晶的温度;在最佳的保留温度条件下,其热滞活性为1.2℃。碱性蛋白酶酶解制备鱼皮抗冻蛋白是一种快速有效的方式,酶解获得的抗冻蛋白热滞活性高。该结果为综合发展和利用金线鱼鱼皮提供一定的理论依据。     

超声波辅助酶解法提取罗非鱼眼透明质酸工艺条件

本文研究超声波辅助酶解法提取罗非鱼眼玻璃体中透明质酸,通过单因素实验、正交实验分析了超声波功率、超声处理时间及蛋白酶种类、酶解时间、酶解pH、酶量、酶解温度对透明质酸提取的影响,确定了最佳提取工艺条件:超声波功率200W、超声处理时间15min、酶作用时间3 h、酶解温度40℃、酶解pH值9.0、酶用量6000U·g-1,利用最优条件透明质酸的平均得率为11.44%。采用季铵盐络合法、离子交换层析法对透明质酸粗品进行纯化得到高纯度的透明质酸,其纯度达到97.47%,利用粘度法测量透明质酸相对分子量为100 KDa。紫外图谱中无蛋白及核酸的吸收峰,红外光谱图显示其特征吸收峰和标准品相同。本研究为罗非鱼眼的高值化利用提供了技术支撑。     

超声辅助竹节虾头酶解及抗氧化肽分离研究

为了实现竹节虾加工副产物的高值化利用,以竹节虾加工废弃物中的虾头副产物为原料,以水解度和DPPH清除率作为评价指标,采用中性蛋白酶酶解,通过响应面法优化超声辅助酶解工艺,并依次通过超滤、凝胶层析色谱和反相高效液相色谱等分离方法,从竹节虾虾头酶解产物中分离制备抗氧化肽,采用超高压液相色谱串联质谱联用技术对肽的结构进行表征。结果表明,在中性蛋白酶添加量为3 000 U·g~(-1)、p H值7.0条件下,最佳超声辅助酶解工艺参数为超声时间41 min,超声温度55℃,超声频率22 k Hz,料液比1∶9(w/v),在此条件下获得的酶解产物DPPH清除率达69.50%。当水解时间为0.5~2.5 h时,超声辅助酶解的酶解产物水解度和DPPH清除率比非超声辅助酶解工艺分别高17.95%和18.83%,该工艺缩短了酶解时间,节约了能耗。酶解产物经超滤初步分离发现,相对分子质量在3k Da(SHP4)的组分具有显著的抗氧化活性。用凝胶层析法进一步分离纯化SHP4组分后得到4个峰,其中SHP4-II的DPPH清除率最高;SHP4-II通过反相高效液相纯化后也得到4个主要肽峰,在多肽含量为1.5 mg·m L~(-1)时,SHP4-II-4的DPPH清除率最高,达到85.69%,并且具有较好的分离度,质谱分析发现,该抗氧化肽的结构为Gly-Asn-Gly-Leu-Pro(455.99 Da)。本研究结果为虾肽抗氧化保健食品的研发提供了一定的科学依据。     

短程分子蒸馏技术精制巨尾桉叶精油工艺优化

为了优化巨尾桉叶精油精制工艺,采用短程分子蒸馏,对初级桉叶精油中1,8-桉叶素和α-蒎烯进行精制,研究不同温度和压力条件下2种物质分离特性。以巨尾桉叶为试验材料,依次采用超临界CO2萃取和分子蒸馏对其进行桉叶油树脂提取和纯化得到初级桉叶精油,采用二因素五水平的响应面优化试验,将馏出物得率、馏出物中1,8-桉叶油素质量分数及α-蒎烯质量分数、馏余物中1,8-桉叶油素质量分数及α-蒎烯质量分数作为试验指标,对分子蒸馏精制桉叶精油工艺进行优化研究。最优纯化工艺条件:以馏出物为目标产物,蒸馏温度38℃,蒸馏压力41 Pa,馏出物中1,8-桉叶油素和α-蒎烯的质量分数分别约为60.80%和31.58%,馏出物的得率为82.06%。分子蒸馏能够对桉叶精油进行有效的纯化精制,桉叶初级精油经过二级分子蒸馏精制后,1,8-桉叶油素和α-蒎烯的质量分数分别提高了77.62%和56.72%。蒸馏温度和蒸馏压力对1,8-桉叶油素质量分数的影响均较α-蒎烯明显,同时,对于1,8-桉叶油素,蒸馏压力的变化对其质量分数的影响较蒸馏温度明显,相反的,对于α-蒎烯,蒸馏温度对其质量分数影响更为显著(P0.05),该研究结果可为分离提纯2种物质提供技术参考。     

超声波辅助酶法提取甘薯渣膳食纤维的研究

为了高效提取薯渣膳食纤维,本文以甘薯渣为原料,采用超声波技术辅助酶法提取甘薯膳食纤维,探讨料水比、超声功率、超声时间、酶用量和酶解温度对甘薯膳食纤维得率和可溶性膳食纤维含量的影响,并采取正交法对提取工艺进行优化。确定甘薯渣膳食纤维的提取条件为:料液比1∶50,超声功率400W,超声时间10min,纤维素酶用量3μL·g-1,酶解温度65℃;该工艺条件下膳食纤维得率为39.17%,可溶性膳食纤维含量高达13.49%。本研究为甘薯的精深加工提供了理论依据,并为进一步研究甘薯膳食纤维的构效关系奠定了基础。     

响应面优化酶解法制备猪肩胛骨抗氧化肽工艺

为有效利用废弃猪骨制备抗氧化肽,以猪肩胛骨蛋白为原料,采用响应面分析法优化酶解猪肩胛骨蛋白制备抗氧化肽的工艺条件。结果表明,以多肽得率和DPPH自由基清除率为评价指标,与未加酶处理组进行对比,筛选出风味蛋白酶为最优酶。在单因素试验的基础上,根据Box-Behnken中心组合试验设计原理,最终确立最优的酶解工艺参数为酶解时间4 h、酶解温度55℃、酶底比6 200 U·g~(-1)、底物浓度3%、pH值6.0。在此条件下DPPH自由基清除率为75.90%,测得的多肽得率为62.80%,其中R_(adj)~2=97.05%,R2=98.95%,表明该模型拟合程度较好。本研究结果对响应面优化酶解法制备猪肩胛骨抗氧化肽工艺具有一定的实践参考意义。     

细点圆趾蟹肉酶解肽螯合锌工艺的优化

为了优化蟹肉酶解肽的锌螯合工艺,以细点圆趾蟹肉为原料,制得蟹肉酶解物,通过单因素试验,考察酶解物与硫酸锌的质量比、pH、反应时间、酶解物浓度、反应温度对锌修饰蟹肉酶解肽的影响,采用正交试验优化蟹肉酶解肽的锌修饰工艺。结果表明,在酶解物与硫酸锌的质量比2∶1、pH值6.5、时间60 min、酶解物浓度5%、温度55℃条件下,酶解肽-锌的螯合率为91.52%,螯合物得率为25.83%,该工艺条件下酶解肽与锌的螯合率和螯合物得率均较高。本研究结果为新型生物态锌的研制、细点圆趾蟹的高值化利用提供了依据。     

中性蛋白酶水解鸡骨泥制备短肽工艺优化

为了探究酶法制备鸡骨泥短肽的最佳工艺,该文研究了以中性蛋白酶酶解鸡骨泥时各因素对短肽得率和羟自由基清除率的影响,以及鸡骨泥肽清除超氧阴离子自由基和1,1-二苯基-2-三硝基苯肼自由基(2,2-diphenyl-1-picrylhydrazyl)的能力。试验探究了酶解过程中不同底物浓度、酶浓度、反应温度和反应时间对短肽得率和羟自由基清除率的影响,采用响应曲面优化得到了以短肽得率为目标的中性蛋白酶酶解鸡骨泥的最优工艺,当反应液p H值为7.2,反应温度42℃,反应时间2 h,底物质量分数5%,酶添加量200 mg/g,该条件下制备鸡骨泥肽,测得其短肽得率为56.16%,羟自由基清除率为54.12%。采用该酶解工艺制备鸡骨泥肽,测得其对超氧阴离子自由基最高清除率为56.01%,对DPPH自由基最高清除率为81.57%,说明鸡骨泥肽具有一定的抗氧化活性,研究结果为酶法制备鸡骨泥肽提供参考。     

尿素包接法预浓缩鱼油生理活性组分的研究

为了提高鱼油中生理活性组分ＥＰＡ和ＤＨＡ的含量，以鱼油乙酯为原料，将其与饱和的尿素－乙醇溶液进行包合作用，而使鱼油中的饱和脂肪酸成结晶析出，以达到预浓缩的目的。随着尿素与鱼油乙酯摩尔比的增加，或结晶温度的降低，容易获得ＥＰＡ和ＤＨＡ含量在７０％以上的制品。     

Food matrix effects on in vitro digestion of microencapsulated tuna oil powder

Tuna oil, containing 53 mg of eicosapentaenoic acid (EPA) and 241 mg of docosahexaenoic acid (DHA) per gram of oil, delivered as a neat microencapsulated tuna oil powder (25% oil loading) or in food matrices (orange juice, yogurt, or cereal bar) fortified with microencapsulated tuna oil powder was digested in simulated gastric fluid or sequentially in simulated gastric fluid and simulated intestinal fluid. The level of fortification was equivalent to 1 g of tuna oil per recommended serving size (i.e., per 200 g of orange juice or yogurt or 60 g of cereal bar). The changes in particle size of oil droplets during digestion were influenced by the method of delivery of the microencapsulated tuna oil powder. Lipolysis in simulated gastric fluid was low, with only 4.4-6.1% EPA and ≤1.5% DHA released after digestion (as a % of total fatty acids present). After sequential exposure to simulated gastric and intestinal fluids, much higher extents of lipolysis of both glycerol-bound EPA and DHA were obtained (73.2-78.6% for the neat powder, fortified orange juice, and yogurt; 60.3-64.0% for the fortified cereal bar). This research demonstrates that the choice of food matrix may influence the lipolysis of microencapsulated tuna oil.     

Flavonoids with potent antioxidant activity found in young green barley leaves

Saponarin, a flavonoid found in young green barley leaves, possesses potent antioxidant activities, which are determined by its inhibition of malonaldehyde (MA) formation from various lipids oxidized by UV light or Fenton's reagent. Lipids used were squalene, ethyl linoleate, ethyl linolenate, ethyl arachidonate, octadecatetraenoic acid (ODTA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), cod liver oil, lecithin I, lecithin II, and blood plasma. The addition of saponarin inhibited the formation of MA from squalene upon UV irradiation at the level of 2 μmol/mL by almost 100%, whereas BHT inhibited its formation by 75% at the same level. Saponarin showed potent antioxidant activity toward fatty acid ethyl esters at levels >100 μg/mL. Saponarin inhibited MA formation in ODTA by 60%, in EPA by 50%, and in DHA by 43% at the level of 15 μmol/mL. Saponarin exhibited strong antioxidant activities with dose-response levels toward cod liver oil and lipoproteins (lecithins I and II), higher than those of α-tocopherol. A mixture of saponarin/lutonarin (4.5:1, w/w) inhibited MA formation appreciably from all lipids tested with dose response. This mixture exhibited highest effect toward cod liver oil (86%), followed by DHA, lecithin II, blood plasma, EPA, and lecithin I. Supplementation of young green barley leaves containing saponarin should be beneficial to health and may prevent diseases caused by oxidative damage such as various cancers, inflammations, and cardiovascular diseases.     

Upgrading of maatjes herring byproducts: production of crude fish oil

Fish oil has been extracted from byproducts of the maatjes (salted) herring production using a pilot plant consisting of a mincer, heat exchanger, and three-phase decanter. The crude herring oil obtained had an initial peroxide value (PV), anisidine value (AV) and free fatty acids (FFA) level of only 3 mequiv of peroxide/kg of lipid, 8.9, and 2.9%, respectively. 5,8,11,14,17-Eicosapentaenoic acid (EPA) and 4,7,10,13,16,19-docosahexaenoic acid (DHA) were present in considerable amounts (99 and 91 g/kg, respectively). During storage of the oil, no photooxidation could be detected. Storage at room temperature led to significant autoxidation over time, apparent from primary and tertiary oxidation products, measured by a decrease of hydroperoxides and an increase of fluorescent compounds (FC). Storage at 50 degrees C resulted in significant increases in secondary (AV) and tertiary oxidation (FC) products. At all storage conditions, the FFA contents remained low (<3%) and the alpha-tocopherol content remained constant. These results open the possibility for fish oil production of good quality using salted herring byproducts.     

杉木油的提取及干馏油的精制

水蒸气蒸馏得到的杉木油品质优良,但得率低,为1.7%,干馏法粗杉木油得率高,为9.6%,但质量差,不能直接用于调香。为了提高杉木油得率和质量,该文以杉木根为原料,分别采用水蒸气法和干馏法制备杉木油,并对干馏油进行精制研究,重点研究过氧化物和脂肪醇存在下的自由基取代反应,及反应条件对干馏油香气改善效果的影响,得出较佳的工艺条件为:反应温度140℃,反应时间1h。采用GC-MS对提取的杉木油化学成分进行鉴定,其中水蒸气杉木油含有26种成分、杉木粗干馏油含有50种成分、干馏精油含有34种成分,它们的主要化学成分基本一致,都含有α-柏木烯、β-柏木烯、柏木醇、罗汉柏烯等体现柏木油特征香气的化学成分。粗干馏油成分复杂,经过精制处理,除去了大量的羧酸类、酚类及α,β-不饱和羰基化合物等影响干馏油香气质量的物质。干馏精油色浅、具有柔和的木香、膏香、树脂样的气息,基本能满足调香的要求,在香料工业中有广阔的应用前景。     

Enzyme-assisted acidolysis of borage (Borago officinalis L.) and evening primrose (Oenothera biennis L.) oils: incorporation of omega-3 polyunsaturated fatty acids.

Lipase-catalyzed acidolysis of borage (Borago officinalis L.) and evening primrose (Oenothera biennisL.) oils with long-chain omega3 polyunsaturated fatty acids (PUFA), namely, eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids, was carried out in hexane, and the products were analyzed using gas chromatography. The most effective lipase for incorporation of omega3 PUFA into these oils was Pseudomonas sp. as compared to lipases from Mucor miehei and Candida antarctica. Response surface methodology was used to obtain a maximum yield of EPA+DHA incorporation while using the minimum amount of enzyme possible. The process variables studied were the amount of enzyme (150-350 units), reaction temperature (30-60 degrees C), and reaction time (6-30 h). All experiments were carried out according to a face-centered cube design. Under optimum conditions, incorporation of EPA+DHA was 35.5% in borage oil and 33. 6% in evening primrose oil. The modified borage and evening primrose oils containing gamma-linolenic acid, EPA, and DHA were successfully produced and may have potential health benefits.     

基于脂肪酸组成特征的沙丁鱼油掺伪检测

为了解决鱼油原料市场日益突出的掺伪问题,本研究采用气相色谱法测定沙丁鱼及5种可能掺伪海水鱼的鱼油脂肪酸组成,并运用主成分分析（PCA）探究鱼油中各脂肪酸的变化规律,筛选出可能掺伪的海水鱼鱼油;运用偏最小二乘判别分析（PLS-DA）和多元逐步线性回归分析建立沙丁鱼油的掺伪鉴别模型。结果表明,5种鱼油与沙丁鱼油的脂肪酸组成有不同程度的差异,其中鳀鱼油与沙丁鱼油脂肪酸组成最为接近。PCA分析的总贡献率为89.8%,说明分析结果具有良好的代表性,鳀鱼油可以拟作沙丁鱼的掺伪油。PLS-DA分析认为沙丁鱼掺伪油的6种特征脂肪酸分别是二十碳五烯酸（EPA）、二十二碳六烯酸（DHA）、棕榈酸、棕榈油酸、二十二碳烯酸和油酸。沙丁鱼油掺伪鉴别模型所得相关系数（R2）分别为0.927、0.963、0.987、0.991,各模型平均误差分别为2.20%、1.38%、1.19%、0.83%,表明模型准确性较好。因此,该模型可实现鳀鱼油掺伪沙丁鱼油的定量识别,在鉴别沙丁鱼油掺伪上具有一定的可行性。     

5种提取方法对甲鱼油品质的影响

为探究酶解法、淡碱水解法、溶剂法、超声辅助溶剂法及蒸煮法这5种提取方法对甲鱼油品质的作用效果,以甲鱼脂肪为原料,采用气相色谱仪(GC)和顶空固相微萃取-气质联用仪(HS-SPME-GC-MS)对甲鱼油中脂肪酸组成及挥发性成分变化进行测定分析,并结合感官评价及理化指标分析。结果表明,5种提取方法对甲鱼油的感官与理化特性、脂肪酸组成及挥发性成分存在显著性差异(P<0.05),其中,超声辅助溶剂法的甲鱼油提取率最高,达78.51%,该方法提取的甲鱼油酸价(0.97 mg KOH·g^-1)、过氧化值(2.16 m Eq·kg^-1)均优于其他方法,并含有丰富的油酸、EPA和DHA。此外,超声辅助溶剂法的甲鱼油壬醛、己醛、2-壬酮、2-十一酮及1-戊烯-3-醇主体特征风味化合物的相对含量均较低,鱼腥味较淡,色泽透亮。综上表明,超声辅助溶剂法提取的甲鱼油品质优于其他提取方法。本研究可为今后优化甲鱼油提取方法提供理论依据。     

Efficient hydrolysis of tuna oil by a surfactant-coated lipase in a two-phase system

A surfactant-coated lipase (SCL) prepared by mixing Candida rugosa lipase with emulsifier in ethanol was used to hydrolyze tuna oil in a two-phase aqueous-organic system. Both enzyme (SCL) and substrate (tuna oil) were soluble in the organic phase, and the hydrolysis could occur with water molecules from the aqueous phase. This hydrolysis could promptly proceed compared to that catalyzed by native lipases which only occurred at the interface between the two phases. Michaelis-Menten kinetics in the two-phase reactions showed that the K(m) value of the SCL was half that of the native lipase, while the maximum velocity (V(max)) was 11.5 times higher. The hydrolysis method resulted in enrichment of n-3 polyunsaturated fatty acid (n-3 PUFA) content in glyceride mixtures from 26.4% to 49.8% and DHA from 19.1% to 38.9%. The SCL acted as an efficient hydrolytic catalyst for tuna oil.     

Quantitative ester analysis in cachaca and distilled spirits by gas chromatography-mass spectrometry (GC-MS)

An analytical procedure for the separation and quantification of ethyl acetate, ethyl butyrate, ethyl hexanoate, ethyl lactate, ethyl octanoate, ethyl nonanoate, ethyl decanoate, isoamyl octanoate, and ethyl laurate in cachaca, rum, and whisky by direct injection gas chromatography-mass spectrometry was developed. The analytical method is simple, selective, and appropriated for the determination of esters in distilled spirits. The limit of detection ranged from 29 (ethyl hexanoate) to 530 (ethyl acetate) microg L(-1), whereas the standard deviation for repeatability was between 0.774% (ethyl hexanoate) and 5.05% (isoamyl octanoate). Relative standard deviation values for accuracy vary from 90.3 to 98.5% for ethyl butyrate and ethyl acetate, respectively. Ethyl acetate was shown to be the major ester in cachaca (median content of 22.6 mg 100 mL(-1) anhydrous alcohol), followed by ethyl lactate (median content of 8.32 mg 100 mL(-1) anhydrous alcohol). Cachaca produced in copper and hybrid alembic present a higher content of ethyl acetate and ethyl lactate than those produced in a stainless-steel column, whereas cachaca produced by distillation in a stainless-steel column present a higher content of ethyl octanoate, ethyl decanoate, and ethyl laurate. As expected, ethyl acetate is the major ester in whiskey and rum, followed by ethyl lactate for samples of rum. Nevertheless, whiskey samples exhibit ethyl lactate at contents lower or at the same order of magnitude of the fatty esters.