Enzyme Modified Cheese Flavour Ingredients

End of Project ReportEnzyme-modified cheeses (EMCs) are defined as concentrated cheese flavours produced enzymatically from cheeses of various ages and are principally used as an ingredient in processed foods, where they provide a cost-effective alternative to natural cheese. They can be used as the sole source of cheese flavour to intensify an existing cheese taste, or to impart a specific cheese character to a more bland product. Their main applications are in processed cheese, analogue cheese, cheese spreads, snack foods, soups, sauces, biscuits, dips and pet foods. Their main advantages over other cheese flavour ingredients are: low production costs, consistency, high flavour intensity, diverse flavour range, extended shelf- life, low storage costs and increased functionality. EMCs are generated utilising the same flavour pathways that occur in natural cheese ripening i.e. proteolysis, lipolysis and glycolysis. They are not as easy to differentiate as natural cheeses, as they are characterised by flavour and aroma alone as texture is not a factor in EMC production. The relationship of the flavour of EMCs to the flavour of the corresponding natural cheese remains unclear. This is especially true for Cheddar EMC which is commercially available in a range of Cheddar flavours . Despite the fact that a wide range of commercial EMCs are available, there is very little detailed information available regarding their properties or the specific production processes used. The main objective of this research was to build a knowledge base on EMC products and to utilise this to develop a biotechnological process for the production of improved enzyme modified cheeses for use as flavour ingredients. The strategy was to establish quantitative relationships between the compositional, proteolytic and lipolytic parameters and the sensory characteristics of EMCs. This data would then be used to develop a predictive model for flavour development in EMC production and the subsequent generation of an optimised EMC process enabling the generation of a range of cheese flavours from single or multiple substrates.Department of Agriculture, Food and the Marin

Symposium review: Genomic investigations of flavor formation by dairy microbiota

peer-reviewedFlavor is one of the most important attributes of any fermented dairy product. Dairy consumers are known to be willing to experiment with different flavors; thus, many companies producing fermented dairy products have looked at culture manipulation as a tool for flavor diversification. The development of flavor is a complex process, originating from a combination of microbiological, biochemical, and technological aspects. A key driver of flavor is the enzymatic activities of the deliberately inoculated starter cultures, in addition to the environmental or “nonstarter” microbiota. The contribution of microbial metabolism to flavor development in fermented dairy products has been exploited for thousands of years, but the availability of the whole genome sequences of the bacteria and yeasts involved in the fermentation process and the possibilities now offered by next-generation sequencing and downstream “omics” technologies is stimulating a more knowledge-based approach to the selection of desirable cultures for flavor development. By linking genomic traits to phenotypic outputs, it is now possible to mine the metabolic diversity of starter cultures, analyze the metabolic routes to flavor compound formation, identify those strains with flavor-forming potential, and select them for possible commercial application. This approach also allows for the identification of species and strains not previously considered as potential flavor-formers, the blending of strains with complementary metabolic pathways, and the potential improvement of key technological characteristics in existing strains, strains that are at the core of the dairy industry. An in-depth knowledge of the metabolic pathways of individual strains and their interactions in mixed culture fermentations can allow starter blends to be custom-made to suit industry needs. Applying this knowledge to starter culture research programs is enabling research and development scientists to develop superior starters, expand flavor profiles, and potentially develop new products for future market expansion

Partitioning of starter bacteria and added exogenous enzyme activities between curd and whey during Cheddar cheese manufacture

peer-reviewedPartitioning of starter bacteria and enzyme activities was investigated at different stages of Cheddar cheese manufacture using three exogenous commercial enzyme preparations added to milk or at salting. The enzyme preparations used were: Accelase AM317, Accelase AHC50, Accelerzyme CPG. Flow cytometric analysis indicated that AHC50 or AM317 consisted of permeabilised or dead cells and contained a range of enzyme activities. The CPG preparation contained only carboxypeptidase activity. Approximately 90% of starter bacteria cells partitioned with the curd at whey drainage. However, key enzyme activities partitioned with the bulk whey in the range of 22%–90%. An increased level of enzyme partitioning with the curd was observed for AHC50 which was added at salting, indicating that the mode of addition influenced partitioning. These findings suggest that further scope exists to optimise both bacterial and exogenous enzyme incorporation into cheese curd to accelerate ripening.Department of Agriculture, Food and the Marin

Free Fatty Acids Quantification in Dairy Products

Quantification of free fatty acids in dairy products is not only important due to their (fatty acids) impact on the flavour and texture of dairy products but also because of their potential impact on nutrition and health, and as anti-microbial agents. This chapter provides an overview of the practical issues associated with existing lipid extraction techniques and quantification procedures using gas chromatography flame-ionization detection. The most widely used methods are compared and recent advancements in the quantification of free fatty acids in dairy products are discussed

Model System for the Production of Enzyme Modified Cheese (EMC) Flavours.

End of Project ReportNatural cheese flavour ingredients, in the form of enzyme modified cheeses (EMCs), are widely used in the convenience food industry and can provide high volume added opportunities for the cheese industry. Many EMCs are produced using commercial enzyme preparations and previous studies have indicated that they contain side activities in addition to their stated main activity (see DPRC Report No.10). Therefore, it is critical that the exact enzyme complement of these preparations are known before they can be used to produce EMC of specific requirements on a consistent basis. The scientific basis of rapid enzyme mediated flavour formation in the production of EMCs is not fully understood. Consequently this knowledge gap is a major obstacle in the development of high value cheese flavour ingredients. Hence, a major objective of this project was to deepen the scientific understanding of flavour formation with a view to the production of natural enzyme-mediated dairy flavour ingredients with commercial potential. The ultimate aim was to develop the technology to produce customised high value dairy flavour ingredients in an optimised process.Dairy LevyDepartment of Agriculture, Food and the Marin

Correlating Volatile Lipid Oxidation Compounds with Consumer Sensory Data in Dairy Based Powders during Storage

peer-reviewedLipid oxidation (LO) is a recognised problem in dairy powders due to the formation of volatile odour compounds that can negatively impact sensory perception. Three commercial dairy powders, fat-filled whole milk powder (FFWMP), skim milk powder (SMP), and infant milk formula (IMF), stored under different conditions (21 °C, 37 °C, or 25 °C with 50% humidity), were evaluated by consumer acceptance studies, ranked descriptive sensory analysis, and LO volatile profiling using headspace solid phase microextraction gas chromatography mass spectrometry (HS-SPME GCMS) over 16 weeks. Significant (p = 0.001) differences in the concentration of LO compounds and sensory perception were evident between sample types in the different storage conditions. The sensory acceptance scores for FFWMP and SMP remained stable throughout storage in all conditions, despite the increased perception of some LO products. The IMF sample was perceived negatively in each storage condition and at each time point. Overall increases in hexanal, heptanal, and pentanal correlated with “painty”, “oxidised”, “cooked”, and “caramelised” attributes in all samples. The concentration of some LO volatiles in the IMF was far in excess of those in FFWMP and SMP. High levels of LO volatiles in IMF were presumably due to the addition of polyunsaturated fatty acids (PUFA) in the formulation

Genetic, enzymatic and metabolite profiling of the Lactobacillus casei group reveals strain biodiversity and potential applications for flavour diversification

Aims: The Lactobacillus casei group represents a widely explored group of lactic acid bacteria, characterized by a high level of biodiversity. In this study, the genetic and phenotypic diversity of a collection of more than 300 isolates of the Lact. casei group and their potential to produce volatile metabolites important for flavour development in dairy products, was examined. Methods and Results: Following confirmation of species by 16S rRNA PCR, the diversity of the isolates was determined by pulsed-field gel electrophoresis. The activities of enzymes involved in the proteolytic cascade were assessed and significant differences among the strains were observed. Ten strains were chosen based on the results of their enzymes activities and they were analysed for their ability to produce volatiles in media with increased concentrations of a representative aromatic, branched chain and sulphur amino acid. Volatiles were assessed using gas chromatography coupled with mass spectrometry. Strain-dependent differences in the range and type of volatiles produced were evident. Conclusions: Strains of the Lact. casei group are characterized by genetic and metabolic diversity which supports variability in volatile production. Significance and Impact of the Study: This study provides a screening approach for the knowledge-based selection of strains potentially enabling flavour diversification in fermented dairy products

A Systems-Wide Analysis of Proteolytic and Lipolytic Pathways Uncovers The Flavor-Forming Potential of The Gram-Positive Bacterium Macrococcus caseolyticus subsp. caseolyticus

main articleMacrococcus caseolyticus subsp. caseolyticus is a Gram-positive, commensal organism documented to be present as a component of the secondary microflora in fermented foods such as Ragusano and Fontina cheeses and Cantonese sausage. In these products, the organism appears to play a role in ripening and the development of the final organoleptic qualities. However, the role of this organism in flavor generation is not well understood. Therefore, the objective of this study was to investigate the role of M. caseolyticus subsp. caseolyticus in flavor compound formation through an examination of enzymatic, metabolomic and genomic data. A bank of M. caseolyticus subsp. caseolyticus strains derived from a variety of niches were examined. Enzyme activities analyzed comprised those of the proteolytic and lipolytic cascades including cell-envelope proteinase (CEP), peptidases, esterases, lipases, aminotransferases and glutamate dehydrogenase (GDH). Strain to strain variation was observed, often associated with niche. All strains, except those isolated from non-dairy sources, demonstrated high CEP activity. Such high CEP activity associated with dairy strains implies the importance of this characteristic in the adaptation of these strains to a dairy-specific niche. However, limited downstream peptidolytic activity, in addition to a limited ability to generate free amino acids (FAA) was observed across all strains, indicating weak ability of this organism to generate amino-acid derived flavor compounds. Interestingly, the strains with high CEP activity also demonstrated high esterase activity and gas chromatography-mass spectrometry (GC-MS) analysis of the volatile compounds produced when these strains were grown in lactose-free milk demonstrated differences in the range and types of volatiles produced. In contrast to this metabolic versatility, comparative genome analysis revealed the distribution of components of the proteolytic and lipolytic system in these strains to be conserved. Overall, this study demonstrates the potential of M. caseolyticus subsp. caseolyticus to generate diverse volatile flavor compounds. Additionally, the identification of the highly active strain-specific cell wall bound caseolytic proteases deriving extensive casein hydrolysis, serves as a promising avenue which can be potentially harnessed in the future to produce greater and more diverse flavor compounds

Innovative strategies to enhance the sensory quality of dry fermented sausages containing lactic ingredients by the addition of exogenous enzymes

This study investigated the impact of the addition of exogenous enzymes (Accelerzyme CPG, Debitrase DBP20) or cellular preparations (FlavoGard), traditionally used in the cheese industry, to accelerate flavour development of dry fermented sausages with 6% of lactic derivatives content. Sausages were fermented to pH 5.0, dried for 32 days and vacuum packed stored under refrigeration for 60 days. Sausages were analysed for physicochemical parameters, technological microbiota and proteolysis after fermentation, drying/ripening and storage. Similar compositional results were obtained in all products (38-39% humidity in the final product; 38.2% fat and 40.7% protein as dry matter throughout the study). Debitrase application positively affected proteolysis by changing the free amino acid profile and increasing non-protein nitrogen and total free amino acids by 2.2 and 11.8-fold, respectively. Accelerzyme increased ripened cheese flavour and overall sensory quality from 5.1 to 5.8; Debitrase increased ripened cheese odour and flavour, bitterness, umami, adhesiveness, pastiness, and overall sensory quality from 5.0 to 5.9, and decreased acid and hardness. This study highlights the effects of adding some exogenous enzyme/bacterial preparations traditionally used in the cheese industry to enhance the flavour of dry fermented sausages with high content of lactic ingredients and increase its sensory quality.info:eu-repo/semantics/acceptedVersio