Lipids in muscles and adipose tissues, changes during processing and sensory properties of meat products

Dry-cured meat products represent a large part of the meat products on the European market. The technologies develop for these products lead to the production of a large scale of meat products with typical sensory traits. Numerous studies have been devoted to optimise the quality traits of these products which are considered as traditional products by the consumer and provide a high added value to the producer. Among the components of the raw material, lipids play a key role in the final quality of these products. Many sensory traits of dry-cured meat products depend on lipid traits of muscle and adipose tissues of fresh meat and on their degradation through a complex set of lipolytic and oxidative reactions during processing. Lipid traits of both muscle and adipose tissues of fresh meat are strongly related to pig rearing conditions, mainly genotype and feeding strategy. During processing, lipids undergo intense lipid hydrolysis controlled by both lipases and phospholipases, which remain active all along the process. Lipids are also subjected to oxidation, which generates numerous volatile compounds. These volatiles contribute to some typical aroma notes of dry-cured meat products such as rancid, aged ham and dry-cured odours. This paper reviews the recent knowledge on the influence of lipid traits of fresh meat, lipid hydrolysis and oxidation on the development of sensory traits of dry-cured meat products.     

SUBCUTANEOUS ADIPOSE TISSUE LIPOLYSIS IN THE PROCESSING OF DRY-CURED HAM

ABSTRACT The activities of subcutaneous adipose tissue lipases and esterases were assayed at different stages (0 to 15 months) in the processing of dry-cured ham. The formation of free fatty acids during the process was also determined. Maximal generation of free fatty acids occurred during the first 10 months. Simultaneously, the triglyceride content decreased while the diglycerides increased during the aging period. Neutral and basic lipases showed maximal activity at the beginning of the process but only neutral lipase remained as the main enzyme responsible for the reported lipolysis during the drying ripening stages. Adipose tissue esterases showed excellent stability but the generation of volatile free fatty acids was negligible, suggesting a minor role of these enzymes.     

The role of muscle enzymes in dry-cured meat products with different drying conditions

Several muscle proteases (cathepsins, calpains, peptidases and aminopeptidases) and lipases (lysosomal acid lipase, acid phospholipase and adipose tissue lipase) are involved in important biochemical mechanisms taking place during the processing of dry-cured meat products which are directly related to the final quality. These enzymes are affected by the conditions typically found in the processing of dry-cured meat products, being dehydration one of the most important factors. This work is presenting the effect of different drying conditions, typical in the processing of dry-cured meat products, on the activity of muscle proteases and lipases as well as its relevance for the final product quality.     

南京板鸭生产过程中脂肪酶和磷脂酶的活力变化研究

研究了板鸭生产过程中各工艺点样品肌肉中酸性脂肪酶、中性脂肪酶和磷脂酶活力的变化趋势。结果显示,板鸭在腌制过程中,这三种酶呈逐渐上升的趋势,而在排坯和风干过程中,酶活性逐渐丧失。通过相关分析表明,板鸭加工过程中,盐含量及水分含量与这三种酶活性之间具有显著的相关性。     

A comprehensive review on molecular mechanism of defective dry-cured ham with excessive pastiness,adhesiveness, and bitterness by proteomics insights

Excessive bitterness, pastiness, and adhesiveness are the main organoleptic and textural defects of dry-cured ham, which often cause a lot of financial losses to manufacturers and seriously damage the quality of the product. These sensory and textural defects are related to the protein degradation of dry-cured ham. Proteomics shows great potential to improve our understanding of the molecular mechanism of sensory and textural defects and identify biomarkers for monitoring their quality traits. This review presents some of the major achievements and considerations in organoleptic and textural defects of dry-cured ham by proteomics analysis in the recent decades and gives an overview about how to correct sensory and textural defects of dry-cured ham. Proteomics reveals that muscle proteins derived from myofibril and cytoskeleton and involved in metabolic enzymes and oxygen transport have been identified as potential biomarkers in defective dry-cured ham. Relatively high residual activities of cathepsin B and L are responsible for the excessive degradation of these protein biomarkers in defective dry-cured ham. Ultrasound-assisted mild thermal or high-pressure treatment shows a good correction for the organoleptic and textural defects of dry-cured ham by changing microstructure and conformation of muscle proteins by accelerating degradation of proteins and polypeptides into free amino acids.     

Contribution of muscle aminopeptidases to flavor development in dry-cured ham

The activity of muscle aminopeptidases (alanyl, arginyl, leucyl and pyroglutamyl aminopeptidases) have been assayed along the processing of dry-cured ham. The generation of free amino acids resulting from aminopeptidase action on N-terminal of proteins and peptides has been also analyzed. The assayed aminopeptidases, except pyroglutamyl aminopeptidase, showed good stability. Alanyl and arginyl aminopeptidases have optimal neutral pH near the pH in ham and, in addition, their spectrum of activity against terminal amino acids is in coincidence with the observed release of free amino acids in ham. So, both aminopeptidases appear to be the main contributors to the generation of free amino acids during the processing of dry-cured ham.     

Muscle and Adipose Tissue Aminopeptidase Activities in Raw and Dry-Cured Ham.

Free amino acids have been analyzed in biceps femoris muscle and adipose tissue from raw and dry-cured ham. A high increase was observed for all amino acids except glutamine and taurine. Major increases were in glutamic acid, arginine, alanine, valine, leucine, and lysine. A survey of five aminopeptidase activities of muscle and adipose tissue from raw and dry-cured ham was performed by using 7-amino-4-methylcoumarin derivatives of five amino acids (Leu, Arg, Ala, Tyr and pGlu) as substrates. Optimum activity was found at neutral pH and around 37°C, except the leucyl hydrolyzing activity which was 45°C. High recoveries of activity (25–75%) were obtained in the dry-cured ham. These enzymes might be responsible for free amino acids increasing during dry-curing.     

干腌火腿中肽的形成机理研究进展

干腌火腿加工过程中,在肌肉中内源酶和微生物的共同作用下,产生多种具有特定生物活性或对产品风味具有提升作用的肽。抗氧化肽、降血压肽和抗菌肽等生物活性肽提升火腿的营养价值;呈味肽如鲜味肽、甜味肽、苦味肽、酸味肽、咸味肽及浓厚感肽,有助于形成火腿独特的风味。该文综述了干腌火腿中肽的功能特性,重点对加工过程中肽的影响因素及形成机理进行介绍,为干腌火腿品质特性提升和新工艺设计提供理论参考依据。     

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.     

Oxidation of microsomal fraction in the processing conditions of dry-cured ham. The in-vitro effect of brine

Throughout the manufacturing process of dry-cured ham intense lipid oxidation occurs. Muscle microsomal membranes were used as a model of muscle oxidation in three different procedures: (i) enzymic reaction; (ii) nonenzymic reaction and (iii) sarcoplasmic proteins and microsomal fraction interaction. Porcine M. Biceps femoris from normal and PSE meat qualities treated with 3% NaCl at different temperatures was used as a model of the dry-cured ham process. M. Biceps femoris from normal porcine meat was used to study the in-vitro effect of brine in the oxidative processes. Results showed an important increase of MDA concentration in enzymic and nonenzymic reactions and a higher than normal oxidation level in PSE meat in samples aged for 6 days at 4 °C. The in-vitro assays showed a high level of nonenzymic lipid oxidation at 3 °C incubation. On the other hand, the enzymic reaction showed greater values of MDA at 20 °C incubation. In-vitro NaCl concentrations seemed to have an antioxidant effect in these conditions. Sarcoplasmic proteins had little effect on the oxidative mechanisms suggesting a lack of interaction of these proteins with the microsomal fraction.     

Lipid and protein oxidation and sensory properties of vacuum-packaged dry-cured ham subjected to high hydrostatic pressure

The effect of HHP treatment (600 MPa) on the oxidative stability of lipids and proteins of vacuum-packaged Iberian dry-cured ham and the impact on the sensory characteristics of the product was investigated. In order to assess how different commercial presentations are affected by HHP treatment, three different presentations of vacuum-packaged Iberian dry-cured ham were considered, namely, (i) intact format (IF) corresponding to non-sliced vacuum-packaged dry-cured ham, (ii) conventional-sliced format (CSF) corresponding to dry-cured ham slices placed stretched out in the package and (iii) alternative-sliced format (ASF) corresponding to dry-cured ham slices piled up horizontally. The oxidation of dry-cured ham lipids and proteins was enhanced by HHP-treatment with the presentation being highly influential on these oxidative reactions. Pre-slicing dry-cured ham results in a more susceptible product to oxidative reactions during pressurisation and subsequent refrigerated storage. Possible mechanisms, by which HHP-induced oxidative reactions would affect particular sensory traits in vacuum-packaged Iberian dry-cured ham such as colour, texture and flavour attributes, are discussed.     

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.     

Contribution of a selected fungal population to proteolysis on dry-cured ham

The proteolytic changes taking place in dry-cured hams lead to increases in free amino acids. Such free amino acids not only contribute to flavour, but also serve as precursors of volatile compounds. Several months of ripening time are required to allow the particular flavour to develop. The fungal population allowed to grow on the surface of some types of dry-cured could play a key role on proteolysis, as it has been shown for dry-cured sausages. The purpose of this work was to study the possible contribution of fungi to proteolysis in dry-cured ham. For this, a strain each of non-toxigenic Penicillium chrysogenum (Pg222) and Debaryomyces hansenii (Dh345), selected for their proteolytic activity on myofibrillar proteins, were inoculated as starter cultures. Changes in the high ionic strength-soluble proteins of an external muscle (adductor) revealed in only 6 months higher proteolysis in the inoculated hams when compared to non-inoculated control hams. Proteolytic strains among the wild fungal population on non-inoculated control hams prevented from obtaining similar differences at the end of processing. However, inoculation with Pg222 and Dh345 led to higher levels for most free amino acids at the external muscle in fully dry-cured hams. In addition, the concentration for some of the more polar free amino acids (i.e. Asp, Glu, Ser and Gln) in inoculated hams was higher at external than at internal (biceps femoris) muscles. These promising results deserve further studies to know the impact of a selected fungal population on the volatile compounds and sensory properties of dry-cured ham.     

Nutritional pork meat compounds as affected by ham dry-curing

This work is focused on the determination of compounds of nutritional interest that are naturally present in pork meat and how they are affected during the processing of dry-cured ham. Such compounds are creatine, creatinine, coenzyme Q₁₀, glutathione, carnosine, anserine, carnitine, taurine, cystine, cysteine and the essential amino acids. Their antioxidant and antyhipertensive functions were evaluated. Of all the assayed substances, only glutathione decreased totally during processing. Carnosine, creatinine, anserine and glutathione showed antioxidant, while cysteine, glutathione and carnosine showed antyhipertensive activity. So, dry-cured ham constitutes an excellent source of essential amino acids (all essential amino acids exhibited a large increase during processing) and other nutritionally interesting compounds such as cystine, cysteine, carnosine, anserine, taurine, carnitine and coenzyme Q₁₀.     

Lipolysis in dry-cured ham maturation

Thirty light Parma hams were tested for muscle lipolytic activity (acid and neutral lipase activity) and free fatty acid (FFA) amounts in M. semimembranosus and biceps femoris, during progressive phases (0, 3, 6, 10 months) of dry-cured ham manufacturing. No correlation was found between the activities of acid and neutral lipases in fresh M. semimembranosus, while during processing the activities were positively related (p<0.1), probably due to effects of muscle composition changes on lipolytic activities. In each processing step tested, acid lipase activities were higher in the M. semimembranosus than in the M. biceps femoris, and FFA amounts varied accordingly, the only exception being for the very dehydrated 10-month old M. semimembranosus, which yielded lower FFA than in the corresponding M. biceps femoris. FFAs in the end product correlated positively with acid and neutral lipase activities of green ham, suggesting that FFA production could be influenced by both raw meat properties and muscle composition during processing.     

Contribution of a selected fungal population to the volatile compounds on dry-cured ham

Dry-cured ham is obtained after several months of ripening. Different fungi strive on the surface, including toxigenic molds. Proteolysis and lipolysis by the endogenous and microbial enzymes seem to play a decisive role in the generation of flavor precursors in dry-cured meat products. In addition, fungi show a positive impact on the volatile compounds of ripened pork loins. However, the contribution of the fungal population to flavor formation in dry-cured ham remains unclear. One selected strain each of Penicillium chrysogenum and Debaryomyces hansenii was inoculated as starter cultures on dry-cured ham. Volatile compounds extracted by solid phase micro-extraction technique were analyzed by gas chromatography/mass spectrometry. A trained panel evaluated flavor and texture of fully ripened hams. The wild fungal population on non-inoculated control hams correlates with higher levels of short chain aliphatic carboxylic acids and their esters, branched carbonyls, branched alcohols, and some sulfur compounds, particularly at the outer muscle. Conversely, P. chrysogenum and D. hansenii seem to be responsible for higher levels of long chain aliphatic and branched hydrocarbons, furanones, long chain carboxylic acids and their esters. The very limited impact of P. chrysogenum on pyrazines in inoculated hams can be due to the activity of the yeast. Lower levels for some of the more volatile linear carbonyls at the ham surface suggest an anti-oxidant effect by micro-organisms. The differences in volatile compounds did not show a neat impact on flavor in the sensorial analysis. Nonetheless, inoculated hams got a better overall acceptability, which has to be attributed to their improved texture. The lower toughness of inoculated hams is a direct consequence of an early settling of a highly proteolytic mold. Thus, the use of selected fungi as starter cultures may be useful to obtain high-quality and safe dry-cured ham.     

生姜蛋白酶和猕猴桃蛋白酶对干腌羊火腿蛋白质降解的影响

为了促进干腌羊火腿肌肉蛋白质降解,加快其风干成熟进程,缩短加工周期,提高羊火腿的品质,在干腌羊火腿中添加生姜蛋白酶和猕猴桃蛋白酶。以带骨鲜羊后腿肉为试验材料,分别设计一个对照组和两个试验组;对照组不采取处理,而试验组分别添加0.05%的生姜蛋白酶和猕猴桃蛋白酶,并在相应条件下进行风干成熟,检测干腌羊火腿的总氮(Total nitrogen,TN)含量、非蛋白氮(Non-protein nitrogen,NPN)含量和蛋白质降解指数(Proteolysis index,PI)等蛋白质降解指标,并通过SDS-PAGE(聚丙烯酰氨凝胶电泳)分析了干腌羊火腿肌肉蛋白质的降解情况。结果表明,与对照组相比,猕猴桃蛋白酶处理组和生姜蛋白酶处理组的TN含量风干成熟后分别增加了1.2倍和1.3倍、NPN含量分别增加1.5倍和1.7倍、PI分别上升1.2倍和1.3倍;SDS-PAGE分析结果表明,生姜蛋白酶降解肌肉蛋白的效率较猕猴桃蛋白酶强。通过蛋白质将指数和SDS-PAGE电泳结果可知,生姜蛋白酶对肌肉蛋白的降解程度比猕猴桃蛋白酶大。     

Effect of prefreezing hams on endogenous enzyme activity during the processing of Iberian dry-cured hams

The use of frozen/thawed raw material in the processing of Iberian dry-cured ham has been studied to determine its effect on the sensory quality of the final product. The proteolysis and lipolysis processes were measured by the proteolytic and lipolytic enzyme activities and free amino acids and free fatty acids. The thawed Iberian hams had lower salt contents throughout the process. The use of thawing raw material did not affect the proteolytic enzymes, cathepsins, aminopeptidases and dipeptidylpeptidases, only the activity of dipeptidylpeptidase III was reduced due to thawing. Moreover, there were no differences in the content of free amino acids between fresh and thawed hams during the whole process. However, the use of thawing hams affected the lipolytic activity. The activity of phospholipase and neutral lipase were significantly higher in the thawed hams and also the content of free fatty acids, at all the stages analyzed. Consumer sensory analysis showed thawed Iberian hams had the lowest hardness, probably due to an intense proteolysis. The acceptability of the Iberian hams was similar between fresh and thawed hams.     

羊肉火腿加工过程中粗肽抗氧化活性

为明确羊肉火腿加工过程中内源抗氧化肽的抗氧化能力变化,本实验以不同加工时期干腌羊肉火腿为研究对象,利用磷酸盐缓冲液提取粗肽并测定多肽质量分数,对粗肽还原力、1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）自由基清除率、2,2’-联氮-二(3-乙基-苯并噻唑-6-磺酸)（2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid),ABTS）阳离子自由基清除率、羟自由基清除率、脂质过氧化抑制率进行评价。结果表明：羊肉火腿加工过程中多肽质量分数显著升高,抗氧化活性显著增强,具体表现为加工过程中还原力显著增强（P＜0.05）,ABTS阳离子自由基和DPPH自由基清除率总体呈上升趋势,分别在成熟结束期和成熟中期表现出最强抗氧化能力（分别为0.09、0.017 8 mmol/L）;羟自由基清除率总体呈下降趋势,在风干期抗氧化能力达到最大。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳结果表明,从原料腿到成熟结束期肌肉中蛋白质逐渐发生降解,产生小分子抗氧化肽。本实验为下一步研究羊肉火腿多肽的抗氧化机制提供参考。     

云南干腌火腿加工工艺及其品质影响因素研究进展

云南干腌火腿是一种重要的传统发酵肉制品,以猪后腿为原料,经腌制、风干、发酵等加工而成。云南干腌火腿主要有宣威火腿、撒坝火腿、三川火腿、老窝火腿、诺邓火腿及鹤庆火腿等,本文对云南干腌火腿加工工艺及干腌火腿品质影响因素进行综述,从原料肉、腌制条件、色泽及风味等几方面做了详细讨论,旨在为干腌火腿品质改进提供一定的理论参考。