Changes of arginyl and leucyl aminopeptidase activities in biceps femoris along Jinhua ham processing

Sixty experimental Jinhua hams were processed by a traditional method. The potential arginyl (RAP) and leucyl (LAP) aminopeptidase activities in biceps femoris were determined. The effects of temperature, salt content, sodium nitrate content and pH value on muscle RAP and LAP activities were evaluated using response surface methodology. Porcine muscle was found to possess very strong potential RAP and LAP activities that decreased gradually during processing from 165,980.97Ug(-1) and 89,201.52Ug(-1) before salting to 3164.70Ug(-1) and 5373.66Ug(-1) after aging, respectively. Temperature, pH value and salt content had significant effects on RAP and LAP activities (P<0.0001). They interacted with each other in their effects on RAP activity (P<0.0001) and influenced LAP activity in exponential way (P<0.05). However, 0-50mgL(-1) sodium nitrate had no detectable effect on either RAP or LAP activity (P>0.05). The regression models showed that muscle RAP and LAP maintained their activities all through Jinhua ham processing. LAP could display very strong actual activity during aging and post-aging, indicating that muscle LAP may play the most important role in generating free amino acids during the processing of Jinhua ham.     

Time-related changes in cathepsin B and L activities during processing of Jinhua ham as a function of pH, salt and temperature

Sixty experimental Jinhua hams were processed by traditional methods using green hams from local crossbred pigs of Lanxi, Zhejiang province, PR China. Biceps femoris was sampled from five hams randomly taken after each processing stage. Potential activities of cathepsin B and L were determined. The effects of temperature, salt content, pH value and nitrate content on the activities of cathepsin B and L were evaluated using response surface methodology (RSM) and the actual activities of cathepsin B and L during Jinhua ham processing were calculated. The results showed that potential activities of cathepsin B and L decreased gradually from 11332.05 to 9955.50 Ug(-1) prior salting to 1055.32-1359.90 Ug(-1) at the end of post-aging, respectively. After processing, about 9.31% original potential activity of cathepsin B and 13.66% original potential activity of cathepsin L were left. Temperature, pH value and salt content showed significant effects on cathepsin B and L activities. By stepwise regression analysis, two quadratic regression equation models were built individually for estimating cathepsin B and L activities. The models predicted that the actual activities of cathepsin B and L were less than 5% of their corresponding potential activities most of the time before half of loft-aging and about 15% and 20% during the rest processing periods. Nevertheless, cathepsin B and L retained actual activities throughout processing, especially during loft-aging and post-aging periods.     

金华火腿加工过程中内源脂肪酶活力变化特点研究

以浙江兰溪两头乌杂交猪后腿为原料,按传统工艺加工金华火腿。分别在原料、腌制结束、晒腿结束、成熟中期、成熟结束、后熟-1和后熟-2七个工艺点随机取5条腿的股二头肌为样品,分别测定中性脂肪酶、酸性脂肪酶的活力变化,以及测定pH5.8时脂肪酶综合活力的变化;同时,利用响应曲面法研究温度、盐含量、硝酸盐含量、亚硝酸盐含量对脂肪酶活力的影响,并利用所得回归方程预测金华火腿加工过程中脂肪酶的实际表现活力。结果表明,加工过程中,脂肪酶活力持续降低,在后熟-1工艺点,酸性脂肪酶和中性脂肪酶活力残留分别是8.16%和3.0%,至后熟-2已测不到酶活;温度、盐含量及其交互项是脂肪酶活力的显著影响因子(P<0.05),硝酸盐和亚硝酸盐含量对脂肪酶活力影响不显著(P>0.05)。加工过程中,预测的脂肪酶实际表现活力在成熟中期工艺点之前低于酸性脂肪酶及中性脂肪酶的潜在活力,之后则高于潜在活力,其变化规律与酸性和中性脂肪酶完全不同。     

Changes of alanyl aminopeptidase activity and free amino acid contents in biceps femoris during processing of Jinhua ham

Sixty experimental Jinhua hams were processed by a traditional method. The potential alanyl aminopeptidase (AAP) activity in biceps femoris was determined. The effects of temperature, salt content, sodium nitrate content and pH on muscle AAP were evaluated using response surface methodology. Porcine muscle was found to possess very strong potential AAP activity that decreased gradually during processing from 201,635 U g⁻¹ before salting to 6147 U g⁻¹ after aging. Temperature, pH and salt content had significant exponential effects on AAP activity (P < 0.001). Both temperature and salt content interacted with pH in their effects on AAP activity (P < 0.01). However, 0–50 mg L⁻¹ sodium nitrate had no detectable effect on AAP activity (P > 0.05). The regression model showed muscle AAP maintaining its activity all through Jinhua ham processing, indicating that muscle AAP may generate free amino acids during the processing and storage of Jinhua ham. The concentrations of free amino acids increased significantly (P < 0.05) during Jinhua ham processing, except for arginine and cystine. The concentrations of most free amino acids were 5–20 times higher in the final product than in hams before salting. Final concentrations exceeded thresholds for sensory detection, thus implicating an important role of free amino acids in the determination of Jinhua ham flavor.     

肌肉非蛋白氮和游离氨基酸在金华火腿加工过程中的变化

以60只浙江兰溪当地杂交猪后腿为原料,按传统工艺加工金华火腿,分析了股二头肌中非蛋白氮(Non-proteinnitrogen,NPN)和游离氨基酸(Freeaminoacids,FAA)浓度随加工时间而变化的规律。结果表明,肌肉中的NPN和FAA含量都随金华火腿加工进程逐渐升高;NPN在后熟期和晒腿期间升高速度最快,而FAA在成熟过程中升高最快,成品火腿的蛋白质降解指数在14～20之间;成品火腿中的FAA占NPN的70%以上,其中以Arg、Glu、Leu、Lys、Ala和Val等含量较高。FAA总含量比腌制前提高13.8倍,火腿中大部分FAA浓度为腌制前的10～20倍,其中增加比例较大的FAA有Lys、Asp、Ser、Tyr和Ile等。火腿中大部分游离氨基酸的浓度多倍于其感觉阈值,因此可能对火腿风味的形成有重要贡献。     

应用响应曲面法研究金华火腿生产过程中磷脂酶的变化

取 6 0条杂交猪后腿作原料按传统金华火腿生产工艺进行生产 ,分别在原料、盐后、晒后、成熟中期、成熟后期、后熟 (1)、后熟 (2 )工艺段随机取 5条腿的股二头肌作为样品测定潜在的磷脂酶活力 ,结果显示潜在的磷脂酶活力在生产过程中不断下降 ,原料中酶活为 3 0 5 ,到后熟 (1)酶活为 0 . 16 ,在后熟 (2 )已测不到酶活。利用响应曲面法研究磷脂酶在生产过程中实际酶活的变化情况并建立酶活变化数学模型 ,结果表明 ,温度和盐含量是影响磷脂酶活力的显著因子 ,通过模型预测 ,生产过程原料中的实际酶活为 0. 2 1,盐后为 0 . 13,之后酶活不断提高 ,到成熟后期达最高 ,为 0 . 4 1,之后再开始下降。     

Dipeptidyl peptidase activities along the processing of Serrano dry-cured ham

The activity of pork muscle dipeptidyl peptidases I, II, III, and IV was followed during the processing of Serrano dry-cured ham. The effects of NaCl, pH, and temperature on these enzymes have also been studied for a better understanding of their importance in ham processing. The lysosomal enzymes DPP I and DPP II showed maximal activity at pH around 5.5-6.0, i.e., close to the pH in ham. At low temperatures, only DPP I and DPP IV showed relevant activity. NaCl considerably inhibited DPP activities except DPP I which was only slightly affected. Dipeptidyl peptidases remained active during the whole process except DPP II whose activity disappeared after 240 days of dry-curing.     

Proteolysis in biceps femoris during Jinhua ham processing

Sixty experimental Jinhua hams were processed by a traditional method. The nitrogen fractions and free amino acids in biceps femoris were analyzed. Intense proteolysis was found in ham muscle and totally more than 10% of muscle proteins were degraded during the course of Jinhua ham processing. The proteolytic index of Jinhua ham was between 14 and 20. Both insoluble and soluble proteins were degraded to some degree and the later showed more intense degradation. In the soluble fraction, the percentage of non-protein nitrogen (NPN) increased gradually whereas that of protein nitrogen decreased during processing (P<0.05). However, very small amount of peptides larger than 1kDa was accumulated during the whole course of processing, which proved that intense degradation reactions were also happened to them, especially at the post ripening stage when dramatic decrease of nitrogen fraction from all the peptides larger than 1kDa was found. As a result, more than 90% of the NPN products from muscle proteolysis were free amino acids and peptides of MW less than 1kDa that might make important contribution to Jinhua ham taste and provide precursors for the generation of volatile flavor compounds in ham muscle. The pattern of muscle proteolysis coincided with the reported changes of muscle proteolytic enzymes during processing, seemingly indicating that the enzymes could have played important roles in ham muscle proteolysis.     

Changes in flavor compounds of dry-cured Chinese Jinhua ham during processing

Sixty experimental Jinhua hams were processed by a traditional method using green hams from local Liangtouwu cross-bred pigs. Samples of Biceps femoris were taken from five hams randomly at each of the key stages of production, i.e., green ham, end of salting, end of sun-drying, middle of loft-aging, end of loft-aging, post-aging-1 and post-aging-2. Analysis of flavor compounds showed 89 compounds at the green ham stage and at the end of salting, 90 at the end of sun-drying, 91 at the middle of loft-aging, 96 at the end of loft-aging, 93 at the post-age-1 and 94 at the post-age-2. The flavor compounds could be clustered in the following chemical families: alkanes and alkenes, aromatic hydrocarbons, alcohols, aldehydes, ketones, carboxylic acids, esters, oxygenous heterocycle compounds, nitrogenous compounds, sulphur compounds, chloride compounds, amides, and terpenes. During processing, the concentrations of aldehydes and carboxylic acids increased; while the concentrations of alcohols, ketones, alkanes, alkenes, aromatic and cyclic hydrocarbons decreased. Sulphur compounds and pyrazines were formed and increased continuously during processing. In the final Jinhua ham, the contents of aldehydes, carboxylic acids, alcohols and ketones were 45.07%, 18.39%, 13.93% and 9.00% of the total flavor substances, respectively. The first principal component was a group dominated by five unbranched aldehydes; the second principal component was a group dominated by five branched and unbranched alcohols, two branched ketones, toluene, hexane and 4-nitrophthalamide. The two principal components explained 90.6% of the total flavor variance in Jinhua ham.     

金华火腿中挥发性风味物质形成过程及变化规律研究

以60只杂交猪后腿为原料,按照传统工艺加工金华火腿,采用固相微萃取(SPME)技术和气谱-质谱联用仪(GC-MS)分离、鉴定技术,研究了金华火腿股二头肌中挥发性风味化合物的形成变化规律。从6个不同加工时期的样品中共鉴定出191种物质,包括43种烃、31种醛、24种酮、21种酯、19种醇、17种羧酸、16种杂环化合物、5种内酯、5种含氯化合物、4种含硫化合物、2种酚、2种酸酐、1种酰胺和1种醚。其中醛类、醇类、烃类、酮类、酸类和酯类化合物含量较高,但各类风味物质的化合物种类和含量的变化规律各不相同。随着加工进行,肌肉中的挥发性化合物种类逐渐增多,但化合物总含量以晒腿结束和成熟中期最高。研究结果表明,与国外干腌火腿相比,金华火腿成品中羧酸、支链化合物、芳香族化合物及杂环化合物种类和含量较多,可能是金华火腿的风味特征。     

金华火腿生产过程中风味成分的变化

取60条杂交猪后腿作为原料，按金华火腿传统生产工艺进行生产，分别在原料、盐后、晒后、成熟中期、成熟结束、后熟（1）、后熟（2）工艺段随机取5条腿的股二头肌作为样品进行风味成分检测．结果显示：原料腿和盐后腿中检出89种风味物质，晒后腿中检测出90种风味物质，成熟中期和成熟结束腿中分别检测出91种和96种风味物质，在后熟（1）和后熟（2）工艺段分别检测出93种和94种风味物质．这些成分可归类为：烷烯烃、芳香烃、醇、醛、酮、酸、酯、含氧杂环化舍物、含氮化合物、含硫化合物、含氯化舍物、酰胺和萜烯类等物质，其生产过程中的变化规律不同．在金华火腿产品的风味成分中，醛类占化舍物总数的45．07％，酸占18．39％，醇占13、93％，酮占9．00％．主成分分析显示，第一主成分主要由5种直链醛组成，第二主成分主要由5种直链醇和支链醇、2种支链酮、甲苯、已烷等组成．第一和第二主成分总计解释了金华火腿风味成分变化总方差的90．6％。     

Influence of partial replacement of NaCl with KCl on formation of volatile compounds in Jinhua ham during processing

The influence of partial replacement of NaCl with KCl on formation of volatile compounds during Jinhua ham processing was evaluated using GC/MS system. Jinhua ham was treated with either 100% NaCl (I) or 60% NaCl and 40% KCl (II). Formation of volatile compounds increased in Jinhua hams during processing for both salt formulations, particularly at the end of the salting period. There were differences in volatile compound formation between formulations I and II after 45 days of processing. Contents of lipid-derived volatiles (hexanal) and Strecker aldehydes (2-methylbutanal and 3-methylbutanal) were higher in Jinhua hams treated with formulation II after 45 days of processing. Partial salt replacement of NaCl with KCl changed formation of volatile compounds in Jinhua hams and may have affected the flavor of finished products.     

Chemical and structural changes in dry-cured hams (Bayonne hams) during processing and effects of the dehairing technique

Pigs of similar genetic backgrounds and feeding regimes were slaughtered in two abattoirs, one carrying out dehairing by scalding and the other by singeing. One ham from each of 80 carcasses was retained. Sixteen fresh hams (8 from each dehairing technique) were used for analysis while 64 hams were processed into dry-cured ham. Sixteen hams (8 from each dehairing technique) were taken for analysis at end of salting (day 14), end of rest (day 78), mid-processing (day 127) and end of processing (day 251). During processing, the water content of all muscles decreased while the salt content increased. The salt concentration in muscle water tended to equalize in all muscles. The nitrogen content of desalted dry matter (i.e. dry muscle tissue) decreased in both Biceps femoris and Semimembranosus. The content of every free amino acid increased with time, except for taurine and glutamine. Electrophoresis of the low ionic strength-soluble fractions showed all protein bands decreased during processing. Electrophoresis of the myofibrillar fractions indicated changes in all bands except actin (42kDa). These changes were more marked in the Semimembranosus than the Biceps femoris in the earlier processing steps. Ultrastructural changes were more marked in Semimembranosus than Biceps femoris. Hardness and chewiness increased in both muscles during the first half of processing then returned to values close to the initial ones in Semimembranosus but changed little in Biceps femoris. The scalded hams lost more weight than the singed ones during processing. The salt content was higher in scalded hams. Water-soluble nitrogen and NPN were higher in singed hams at the end of processing. The scalded hams were saltier and pungent. They had more pronounced aromas of dry ham, rancidity and hazelnut, and less aroma of fresh meat. Their texture was drier and less mellow.     

原料腿对金华火腿腌制及产品品质的影响

采用传统工艺和新工艺加工金华火腿,研究了原料腿重量、鲜冻状态、皮下脂肪厚度对金华火腿腌制过程和加工品质的影响。结果表明:原料腿重影响腌制用盐量、加工各阶段后的失重率、后熟后的产品质量;皮下脂肪厚度影响腌制进程和腌制失重率;冻腿可以作为金华火腿加工的原料腿,但产品加工质量略差。     

Lean colour of green and matured Parma hams: Comparative evaluation and technological relevance of sensory and objective data

Research has been conducted on the relationship between sensory evaluation and objective measurement of pork colour. The study also investigated the relationship between the colour of fresh meat and the colour of matured hams. Sensory evaluation was carried out at 24 h post mortem on the freshly cut surfaces of green hams trimmed for Parma ham production. Objective measurements were performed at the same time on Semimembranosus and Biceps femoris muscles with a Minolta Chromameter Reflectance II CR200/08. Cold carcass weight, lean content, trimmed ham weight, 45 min and 24 h post mortem pH of Semimembranosus muscle were also recorded. The hams were processed according to Parma ham technology. Weight losses at standard processing times were collected. At the end of processing a sample of about one-third of the hams was evaluated for internal lean colour and lean firmness both by sensory panel and objective measurements. Proximate composition was also determined. The results showed highly significant relationships between colour sensory scores and the coordinates L (?), b (?) and hue of the C.I.E. L (?)a (?)b (?) colour system. Among the other parameters measured on fresh meat only 24 h pH was related at a very significant level with sensory and objective colour values. Correlation coefficients were generally higher for Semimembranosus than Biceps femoris. Matured ham colour was linked with fresh meat sensory judgements, L (?), hue and a (?) values. The relationship, though, was not sufficiently strong for use in predicting the colour of matured hams from green ham characteristics. Processing appears to have a positive effect on the colour of pale meat. Weight losses were mainly linked with lean meat content; other quality attributes being less important. Lean firmness of matured hams was related to the colour and pH of the fresh meat.     

Biochemical changes during processing of traditional Jinhua ham

Jinhua ham is the most famous traditional meat product of China and one of the most famed dry-cured hams in the world. Its processing consists of six stages: green ham preparation, salting, washing and sun-drying and shaping, ripening, and post-ripening. Intense proteolysis and lipolysis occur during processing period. As a result, the content of free amino acids in final ham products is 14-16 times that of green ham, and 191 volatile compounds have been identified during processing, which make a major contribution to the flavor of Jinhua ham.     

金华火腿特征风味物质研究进展

金华火腿是我国传统的干腌火腿,以其特殊的风味而著称。通过对金华火腿及其它干腌火腿主要风味物质、形成机理、提取及鉴定方法等研究现状的综述,表明对金华火腿特征风味物质的深入研究有利于了解风味物质形成规律,为金华火腿加工工艺控制及火腿调味料开发应用提供理论基础。     

不同等级金华火腿风味特点研究

随机取5条不同等级金华火腿的股二头肌为样品,利用固相微萃取(SPME)和GC/MS系统进行风味成分研究。结果发现,四级火腿中共检测到116种成分,在1～4级火腿中分别检出84、84、83和92种成分。这些成分可归类为烷烯烃、芳香烃、醇、醛、酮、酸、酯、萜烯类、含氧杂环化合物、含氮杂环化合物、含硫化合物、酰胺类物质和胺类物质。主成分分析显示,第一主成分主要由丁酸、3-甲基丁酸、苯酚、丙酸和丁酸乙酯组成,解释了不同等级金华火腿风味变化总方差的71.6%。第二主成分主要由:已醛、戊醛、2-甲基丁醛、3-甲基丁醛、1-辛烯-3-醇、丙酮和2,3-丁二醇组成,解释了不同等级火腿风味变化总方差的20.5%。根据风味成分产生途径,可以认为微生物是影响产品级别的第一要素。     

不同地域典型干腌火腿肌原纤维蛋白的氧化特性及体外消化性对比

为比较几种典型干腌火腿的可消化吸收性,以巴马火腿、金华火腿、宣威火腿、帕尔玛火腿为原料,研究4 种干腌火腿的肌原纤维蛋白表面疏水性、巯基和二硫键含量、超微结构、蛋白降解程度、消化前后粒径及体外消化率。结果表明：金华火腿肌原纤维蛋白的表面疏水性显著高于巴马、宣威、帕尔玛火腿（P＜0.05）。微观结构分析显示金华火腿肌原纤维蛋白交联和聚集程度最低,肌原纤维排布更为伸展;帕尔玛火腿聚集程度最大,呈现明显聚集状。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳谱图显示金华火腿肌原纤维蛋白在体外消化过程中大幅降解,促进了肌原纤维蛋白粒径的降低。体外模拟消化结果表明,经胃蛋白酶、胰蛋白酶及α-凝乳蛋白酶作用后,金华火腿肌原纤维蛋白水解速率最高（P＜0.05）;帕尔玛火腿肌原纤维蛋白水解速率最低（P＜0.05）。这表明疏水基团的暴露和蛋白结构的伸展为消化酶提供了更多的识别位点,促进了金华火腿中肌原纤维蛋白的水解。综上,肌原纤维蛋白的氧化程度显著影响蛋白的体外消化率。     

Proteolysis,protein oxidation and protease activity in dry‐cured Xuanwei ham during the salting stages

Twenty‐four experimental dry‐cured Xuanwei hams were salted using a standard method for 90 days. The proteolysis, protein oxidation and protease activities in biceps femoris (BF) and semimembranoesus (SM) muscles of dry‐cured Xuanwei ham were investigated during the salting phase. At the end of salting, the salt content increased to 35.2 g kg^?1 muscle in BF and 54.2 g kg^?1 muscle in SM. During the salting stage, salt soluble proteins were degraded mainly into water soluble proteins that were further broken down to peptides with molecular weights mostly greater than 1 kDa. Although large amounts of smaller peptides and free amino acids were generated, especially when the hams were aged. The carbonyl contents were increased but lower than 1.57 nmol mg^?1 proteins in muscles during the salting stage. The cathepsin B, dipeptidyl peptidase I (DPP I), alanyl (AAP), arginyl (RAP) and leucyl (LAP) aminopeptidase all remained active while salt content strongly inhibited cathepsin L and DPP IV in the first 90 days. The results suggested that the salting process promoted the hydrolysis of proteins, and increased the muscle protein oxidation at a slower rate.