Oxidation of sarcoplasmic proteins during processing of Cantonese sausage in relation to their aggregation behaviour and in vitro digestibility

The physicochemical changes of sarcoplasmic proteins, especially oxidation behaviour, were measured to determine their mechanism of action on in vitro protein digestibility during Cantonese sausage processing. The results indicated that carbonyl level increased (p<0.05) during the process. The fluorescence loss of tryptophan residues was a direct consequence of the oxidative degradation. All the parameters of protein aggregation were highly (p<0.05) correlated with carbonyl level and protein surface hydrophobicity (H(0)), indicating that protein oxidation and thermal denaturation could induce protein aggregation, leading to secondary structural changes. The analysis of in vitro digestibility showed no correlation between pepsin activity and protein oxidation, due to the biphasic response of sarcoplasmic proteins toward proteolysis. However, a highly significant (p<0.05) correlation was observed with trypsin and α-chymotrypsin activity, indicating that protein oxidation induced the changes in H(0), protein aggregation and secondary structure, which further influenced in vitro digestibility.     

Physicochemical changes of myofibrillar proteins during processing of Cantonese sausage in relation to their aggregation behaviour and in vitro digestibility

The physicochemical changes of myofibrillar proteins, especially oxidation behaviour, were measured to determine their mechanism of action on in vitro protein digestibility during Cantonese sausage processing. The results indicated that the carbonyl level significantly increased (p < 0.05) during the process. The SH group level decreased, while S–S group level increased gradually. Protein aggregation was induced by oxidation and heat treatment. Result from Fourier transform infrared (FTIR) spectroscopy confirmed protein aggregation occurred. The analysis of in vitro digestibility showed a highly significant (p < 0.05) correlation between pepsin activity and carbonyl group formation, S–S group level, protein surface hydrophobicity, D_4,3. A negative and highly significant correlation between trypsin, α-chymotrypsin activity and carbonyl group formation was measured, while no significant correlation with S–S groups, protein surface hydrophobicity, D_4,3 was observed. It indicated that not only protein oxidation and aggregation but also degradation by pepsin would influence proteolysis with trypsin and α-chymotrypsin.     

Changes in lipid composition,fatty acid profile and lipid oxidative stability during Cantonese sausage processing

Lipid composition, fatty acid profile and lipid oxidative stability were evaluated during Cantonese sausage processing. Free fatty acids increased with concomitant decrease of phospholipids. Total content of free fatty acids at 72 h in muscle and adipose tissue was 7.341 mg/g and 3.067 mg/g, respectively. Total amount of saturated, monounsaturated and polyunsaturated fatty acids (SFA, MUFA, and PUFA) in neutral lipid exhibited a little change during processing, while the proportion of PUFA significantly decreased in the PL fraction. The main triacylglycerols were POO + SLO + OOO, PSO (P = palmitic acid, O = oleic acid, L = linoleic acid, S = stearic acid), and a preferential hydrolysis of palmitic, oleic and linoleic acid was observed. Phosphatidylcholines (PC) and phosphatidylethanolamines (PE) were the main components of phospholipids and PE exhibited the most significant degradation during processing. Thiobarbituric acid values (TBARS) increased while peroxide values and hexanal contents varied during processing.     

Structural characteristics of peptides extracted from Cantonese sausage during drying and their antioxidant activities

Two peptide fractions (P1 and P2) were isolated from Cantonese sausages at different drying periods by ultrafiltration. P1 possessed a stronger 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity in the first 18 h of drying and a weaker activity during the rest period, comparing with P2. The analysis of amino acid composition indicated that histidine was the major amino acid in both peptide fractions, which content was changed during the process. In addition, Glu, Gly, Arg were important constituent amino acids in both peptide fractions. A significant (P < 0.05) decrease of >10 kDa peptide fraction and increases of 2–3 and <2 kDa peptide fractions were detected when comparing with peptides at 0 h. However, no significant difference was found for all the peptide fractions at different drying periods, except for 2–3 kDa peptides. A significant (P < 0.05) correlationship was found between lipid oxidation and antioxidant activities.

Industrial relevance

Cantonese sausage has gained much popularity and acceptance in China and many other countries. In this work, two peptide fractions were isolated from Cantonese sausage. A good antioxidant activity was observed for each peptide fraction. Endogenous peptides resulting from proteolysis offer a promising approach to improve lipid stability for meat industry. It is helpful to retard lipid oxidation in order to improve its quality.     

Chemical oxidation decreases proteolytic susceptibility of skeletal muscle myofibrillar proteins

The objective of this study was to investigate the effect of chemical oxidation on proteolysis susceptibility of myofibrillar proteins. Myofibrils were prepared from pig M. longissimus dorsi and oxidised by a hydroxyl radical generating system. Protein oxidation level was measured by the carbonyl content, free thiol group content and bityrosine formation. Oxidised or non-oxidised myofibrillar proteins were exposed to papain and proteolysis was estimated by fluorescence using fluorescamine. Oxidation of myofibrillar proteins was dependent upon the oxidising agent concentration. Disulfide bridge and bityrosine formation indicated that oxidation by OH° can induce protein polymerization. Electrophoretic study showed that myosin was the protein most sensitive to oxidation. Results showed a direct and quantitative relationship between protein damages by hydroxyl radical and decreased proteolytic susceptibility. Electrophoretic observations suggest that polymerization and aggregation may explain in part decreased susceptibility of myofibrillar proteins to proteolysis.     

Volatile compounds of Cantonese sausage released at different stages of processing and storage

The volatile compounds of Cantonese sausage at different stages of processing and storage were analysed by gas chromatography/mass spectrometry (GC/MS) with solid-phase microextraction (SPME). One hundred and four volatile compounds were identified. Esters and alcohols were the most important chemicals in all the samples, accounting for 16.94–50.12% and 30.01–65.54% of the total area, respectively. Ethanol and its alcohol esters were the major volatile compounds of Cantonese sausage, which suggested that the alcohol added to sausage at the preparation stage was the most important factor for the characteristic flavour of Cantonese sausage. Aldehydes (especially hexanal) contributed between 3.33% and 17.42% to the total areas. The presence of carboxylic acids in esters indicated that lipid oxidation played a key role on the flavour formation of Cantonese sausage. Amino acid catabolism and microbial activity also played an active role in the formation of volatile compounds.     

红曲菌发酵对广式腊肠颜色和蛋白质降解的影响

采用GIM3.439紫红曲菌发酵广式腊肠,研究红曲菌发酵对广式腊肠的色泽、p H、氨基酸态氮含量和感官品质的影响;运用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和高效液相色谱（HPLC）等方法对广式腊肠中蛋白质、肽、游离氨基酸进行检测,分析红曲菌发酵过程中广式腊肠蛋白质的降解,初步研究了红曲菌发酵对广式腊肠中游离氨基酸的影响。结果表明:红曲菌发酵能够加深广式腊肠的红色;在发酵6 h时,发酵组氨基酸态氮含量达0.14%;电泳结果表明,红曲菌发酵对水溶性和盐溶性大分子蛋白降解均有促进作用,能够增加产品中小分子肽及游离氨基酸的总量;游离氨基酸总量由580 mg/100 g增加到930 mg/100 g。红曲菌发酵不仅使产品色泽更加诱人,而且使发酵后腊肠滋味更加丰富、浓郁,提高了腊肠的感官品质和营养价值。      

添加葡萄球菌和微球菌对广式腊肠品质的影响

本研究将从广式腊肠中依据其蛋白酶、脂肪酶和亚硝酸盐还原酶活性筛选出来的葡萄球菌和微球菌2株菌株应用到腊肠中,通过色差计及紫外图谱扫描、不同蛋白组分含量、脂肪氧化、蛋白氧化和色素氧化值测定及感官评价来研究其对广式腊肠色泽、质构、风味及贮藏期氧化稳定性的影响。研究表明:微生物的添加显著提高了a*值及亚硝基肌红蛋白含量。与对照组相比,接菌组腊肠中肌浆蛋白和肌原纤维蛋白含量显著增加,不溶性蛋白减少,从而引起接菌组腊肠硬度值和咀嚼性显著降低,另外接菌组比对照组具有更低的硫代巴比妥酸(TBARS)值、羰基值及高铁肌红蛋白含量和更高的感官评分。因此合适发酵剂的使用有助于色泽和质构的改善,氧化稳定性的提高,从而提高广式腊肠品质及延长货架期。      

低酸度川味香肠加工过程中脂肪降解和氧化

在控温控湿条件下接种发酵剂生产低酸度川味香肠,通过测定加工过程中p H、酸价、过氧化值、TBA值和游离脂肪酸组成,并与自然条件下生产的传统川味香肠(对照组)相比较,揭示低酸度川味香肠加工过程中脂肪降解和氧化的规律。实验结果显示,干燥结束时,低酸度川味香肠的p H、过氧化值和TBA值分别为5.50、0.783meq/kg和0.252mg丙二醛/kg,均低于对照组,低酸度川味香肠酸价为2.701mg KOH/g,高于对照组,低酸度川味香肠的单不饱和脂肪酸含量为40.46%,不饱和脂肪酸含量为59.09%,其不饱和脂肪酸含量明显高于对照组(41.99%)。研究结果表明,在整个加工过程中,低酸度川味香肠中的脂肪更容易发生降解,多不饱和脂肪酸更容易产生氧化现象,脂肪氧化在加工过程中持续进行。      

桑椹多酚对广式腊肠贮藏过程中脂质降解和氧化特性的调控作用

脂质氧化和水解是形成腊肠特征风味的重要途径,但过度氧化会造成腊肠品质的劣变。桑椹多酚是一类天然抗氧化剂,被证实可以有效缓解腊肠储藏过程中的氧化反应,然而其通过调控腊肠脂质氧化和水解来改变腊肠风味的相关机制还鲜有研究报道。鉴于此,该研究将桑椹多酚提取物（1 g/kg）加入到肉糜中,制成广式腊肠,跟踪28 d的储藏过程中腊肠的脂质氧化代谢产物（共轭二烯、共轭三烯、TBARS和己醛）以及脂肪酸（中性脂肪酸、游离脂肪酸和磷脂）的组成变化。研究发现,桑椹多酚主要通过抑制脂质次级代谢产物（己醛和丙二醛）来延缓腊肠的脂质氧化。经过28 d的储藏期后,桑椹腊肠中己醛与丙二醛的含量较对照组分别下降63.10%与28.05%。此外,储藏过程中桑椹多酚还可以有效抑制腊肠储藏过程中游离脂肪酸、磷脂的释放。经过28 d的储藏期后,与对照组相比,桑椹腊肠的游离脂肪酸含量下降了9.59%。同时,桑椹多酚还可以减少风味物质中醛类物质的产生和积累。综上所述,桑椹多酚主要通过延缓腊肠储藏过程中脂质的降解及次级氧化过程来调控产品的风味。     

亲水性胶体对广式腊肠水分迁移及质构的影响

本文研究了卡拉胶、魔芋胶、瓜尔豆胶及其复配对广式腊肠烘烤过程中水分迁移、产品质构、感官等质量品质的影响。结果表明,广式腊肠添加亲水性胶体后,可略有提高产品的水分含量,尤其是瓜尔豆胶组(19.94%),比对照提高4.02%;在烘烤中期,亲水性胶体可降低广式腊肠的干燥速率,整体而言,亲水性胶体对烘烤过程中水分散失影响较小。胶体添加组感官得分除D3组,均高于对照组,其中添加复配卡拉胶与魔芋胶的感官综合得分最高,达到87.5;除瓜尔豆胶外,单独胶体和复配胶均能一定程度上改善广式腊肠的质构特性,尤其是复配胶体组。广式腊肠添加亲水性胶体后,复水率显著提高(p0.05),尤其是瓜尔豆胶及其复配组,达到3.79%。复配胶的加入可改善广式腊肠的感官品质、质构特性及复水性,不同亲水性胶体有不同的作用特点。     

广式腊肠加工过程中肠球菌变化规律及其分离鉴定

本文对广式腊肠烘烤加工过程中肠球菌的变化规律进行了研究,结果发现:肠球菌在烘烤0-18 h时数量增加,烘烤18,,-54h数量下降,烘烤54h后数量缓慢提高,至烘烤结束时肠球菌数量为2.74logcfu/g,占乳酸菌总数的0.04%,为次要茵群.分离得到的两株肠球菌AD4和AD8经鉴定为屎肠球菌.AD4和AD8能够在50℃生长以及耐受6.5%NaCl,因此能够适应广式腊肠的环境;AD4和AD8均不具有明胶酶活性,但都具有溶血性,表明广式腊肠中的肠球菌具有某些毒力因子.     

中式香肠风干过程中蛋白氧化对蛋白聚集、水解性及溶解性的影响

从中式香肠中提取肌浆蛋白和肌原纤维蛋白,并测定其羰基值、巯基值和蛋白浊度,以及蛋白质降解情况和溶解性,确定蛋白氧化对蛋白聚集性、水解性和溶解性的影响。研究结果表明:蛋白羰基含量都显著增长,巯基含量显著下降。蛋白氧化导致蛋白溶液浊度上升,蛋白表面疏水性增加,以及蛋白发生不同程度的降解。蛋白溶解性实验表明氢键、疏水作用力、二硫键和离子键是形成香肠蛋白三维网状结构的主要作用力。     

基于电子鼻和神经网络对广式香肠脂肪氧化的评价

采用电子鼻技术对广式香肠加工和贮藏过程中的脂肪氧化程度进行检测。在加工和贮藏中,分别提取香肠烘干0～120 h和贮藏0～20 周的电子鼻响应值,同时检测香肠的酸价（acid value,AV）和过氧化值（peroxidevalue,POV）来评价香肠的脂肪氧化程度,建立两者之间相关性。通过Loading分析、方差分析和Pearson相关性分析评价10 个传感器对香肠气味的贡献率,选出最佳传感器阵列,通过人工神经网络方法建立香肠AV和POV的预测模型。结果表明：S4、S6、S7、S8和S9为香肠加工过程中对脂肪氧化表征的最佳传感器阵列,S6、S7、S8和S9为香肠贮藏过程中的最佳传感器阵列。在加工和贮藏过程中模型预测效果较好。其中,对于加工过程,基于全部传感器阵列模型对AV和POV预测的R2分别为0.959和0.930,而基于最佳传感器阵列对AV和POV预测的R2分别为0.930和0.914;对于贮藏过程,基于最佳传感器对POV预测模型R2为0.805外,其余皆在0.9以上。因此,电子鼻在广式香肠加工和贮藏过程中对其脂肪氧化程度的检测有着比较好的效果,可以进一步应用到广式香肠的商业生产和贮藏。     

Effect of starter culture on proteolytic changes during processing of fermented beef sausages

Fermented beef sausages inoculated with four different starter cultures (Pediococcus acidilactici,Lactobacillus curvatus, Lactobacillus sake, or Streptomyces griseus) were evaluated for proteolysis during process stages (prefermentation, fermentation, drying and heating). Increases (p ? 0.05) in the nonprotein nitrogen (NPN) fraction were found at sequential stages of processing, while starter cultures had no major effects on NPN content. Concentrations of most free amino acids increased (p ? 0.05) during fermentation and drying, and culture effects were found for differences among concentrations of some individual free amino acids. From SDS–PAGE analysis of sarcoplasmic and myofibrillar protein fractions after fermentation and drying, myosin heavy and light chains, actin and troponin were degraded during processing. However, starter culture effects were absent from SDS–PAGE protein patterns.     

Effect of sage (Salvia officinalis) on the oxidative stability of Chinese-style sausage during refrigerated storage

The objective of this study was to assess the effect of sage, at levels of 0.05%, 0.1% and 0.15% (w/w), on the oxidative stability of Chinese-style sausage stored at 4 °C for 21 days. The results showed that inclusion of sage in sausages resulted in lower L* values (P < 0.05) and higher a* values (P < 0.05) compared to the control. During refrigerated storage, sausages containing sage showed significantly retarded increases in TBARS values, and in the formation of protein carbonyls (P < 0.05), but showed accelerated losses of thiol groups (P < 0.05). Addition of sage to the sausages at levels of 0.1% and 0.15% reduced textural deterioration during refrigerated storage (P < 0.05). Sage used in this study had no negative effects on the sensory properties of sausages.     

Changes in lipid composition and fatty acid profile of Nham,a Thai fermented pork sausage,during fermentation

Changes in lipid composition and fatty acid profile of Nham during fermentation were investigated. Total lipids of Nham were in the range 2–3%. The extracted lipid of initial Nham mix consisted mainly of triglycerides (TG), accounting for more than 75% of the total lipid, followed by phospholipids (PL) and a trace amount of diglycerides (DG) and free fatty acid (FFA). During fermentation, TG, DG and PL decreased with a concomitant increase in FFA, indicating lipolysis of Nham lipids during fermentation. Changes in fatty acids of the total lipids, non-polar and polar lipid fractions were observed during fermentation. In both total and non-polar lipid fractions, the major fatty acids found in a descending order were oleic (C18:1), linoleic (C18:2) and palmitic (C16:0) acids, which together accounted for 90% of the total fatty acids. Increases in fatty acid contents in both total and non-polar lipid fractions, were observed with a corresponding decrease in the quantity of fatty acids of phospholipids. As the fermentation proceeded, peroxide value generally increased while TBARS values decreased. Overall, lipid oxidation in Nham occurred during fermentation but did not cause the objectionable odour and taste in any Nham tested.     

糖添加量对广式腊肠脂质氧化稳定性及感官品质的影响研究

研究了不同糖添加量(3%、6%、9%、12%,m/m)对广式腊肠脂质水解酶活、脂质氧化稳定性及产品感官品质的影响规律。结果表明中性脂肪酶、酸性脂肪酶和磷脂酶活力在加工过程中呈下降趋势(p0.05)。中性脂肪酶活力高于酸性脂肪酶和磷脂酶活力。糖添加量显著影响三种脂肪酶活力在加工过程中的变化,高糖添加组的三种脂肪酶活力略高于低糖添加组;糖添加量为3%时,烘烤结束后中性脂肪酶、酸性脂肪酶和磷脂酶活力分别下降77%、81%和96%,而添加量为12%时,其分别下降67%、65%和78%。烘烤结束后低糖添加组的过氧化值、羰基值和己醛含量较高,表明糖添加量的降低导致脂质氧化的加剧。感官分析表明糖添加量的降低致使咸味更加突出且出现一定的脂质哈败味,降低了广式腊肠产品的接受度。     

洋葱皮提取物对西式香肠抗氧化及品质的影响

通过测定酸价、过氧化值与硫代巴比妥酸值来监测西式香肠在储存过程中的脂肪氧化程度的变化,并采用色差、全质构分析等方法将洋葱皮提取物香肠与空白对照样品的品质进行比较,以考察洋葱皮提取物对西式香肠的色泽、质构以及氧化稳定性的影响。结果显示:洋葱皮提取物的添加量为0.5%时,不仅抑制了西式香肠脂肪的氧化,而且还改善了其色泽,降低了其硬度、弹性与咀嚼性。