Enzymes In Food Processing Research Articles

Bioactive compounds from food-grade Bacillus.

Bacillus species have attracted significant attention in recent years due to their potential for producing various bioactive compounds with diverse functional properties. This review highlights bioactive substances from food-grade Bacillus strains and their applications in functional foods and nutraceuticals. The metabolic capacities of Bacillus species have allowed them to generate a wide range of bioactive substances, including vitamins, enzymes, anti-microbial peptides, and other non-ribosomal peptides. These substances have a variety of positive effects, including potential cholesterol-lowering and immune-modulatory qualities in addition to anti-oxidant and anti-bacterial actions. The uses and mechanisms of action of these bioactive chemicals can be used to improve the functional qualities and nutritional profile of food products. Examples include the use of anti-microbial peptides to increase safety and shelf life, as well as the use of Bacillus-derived enzymes in food processing to improve digestibility and sensory qualities. The exploitation of bioactive compounds from food-grade Bacillus strains presents a promising frontier in the development of functional foods and nutraceuticals with enhanced health benefits. Due to their wide range of activity and applications, they are considered as important resources for the development of novel medications, agricultural biocontrol agents, and industrial enzymes. Ongoing research into the biosynthetic pathways, functional properties, and applications of these compounds is essential to fully realize their potential in the food industry. This review underscores the significance of various bioactive compounds generated from Bacillus in tackling global issues like environmental sustainability, sustainable agriculture, and antibiotic resistance. Future developments in microbiology and biotechnology will enable us to fully utilize the potential of these amazing microbes, resulting in novel approaches to industry, agriculture, and health. © 2024 Society of Chemical Industry.

Enhancing the thermostability of transglutaminase from Streptomyces mobaraensis based on the rational design of a disulfide bond

Transglutaminase (TGase), a transferase, is widely adopted in the food industry and other biological fields due to its unique characteristics of modifying proteins by intra- or intermolecular cross-linking. However, obtaining a mutant TGase that is highly thermostable and active would significantly aid in food processing. Therefore, this study sought to improve the thermostability of TGase by introducing an artificial disulfide bridge through a structure-based rational enzyme engineering approach. After the rational screening, six disulfide mutants (E139C/G143C, R146C/E149C, A182C/N195C, L200C/R208C, T223C/F226C, and E139C/G143C+L200C/R208C) of the transglutaminase gene from Streptomyces mobaraensis (Sm-TGase) were selected and constructed by rationally designed mutations in cysteine. Of them, a mutant (E139C/G143C) with enhanced thermostability was selected and characterized for further analysis. The results indicated that the mutant E139C/G143C had a similar specific activity, optimal temperature, and pH but a lower Km and higher Vmax than the wild-type. Its half-life (t1/2) at 55 °C was 10.7 min, which was 1.69-fold higher than the wild-type, while its melting temperature (Tm) was 3.52 °C higher than the wild-type. These results proved that the introduction of disulfide bonds into TGase by rational design could be an effective approach to improve the thermostability of TGase and other food enzymes for food processing.

A complete workflow for discovering small bioactive peptides in foods by LC-MS/MS: A case study on almonds

Identification of bioactive peptides is an increasingly important target for food chemists, particularly in consideration of the widespread application of proteolytic enzymes in food processing. Because the characterization of small peptides by LC-MS/MS is challenging, we optimized a dimethyl labeling technique to facilitate small peptide identification, using almond proteins as a model. The method was validated by comparing the MS/MS spectra of standards and almond-derived peptides in their nonderivatized and derivatized forms. Signal enhancement of a1 ions was proved to effectively aid in the full-length sequencing of small peptides. We further validated this method using two industrially-relevant protein-rich extracts from almond flour: 1737 medium-sized peptides (5–39 amino acids) and 843 small peptides (2–4 amino acids) were identified. The use of an online bioactive peptide database, complemented by the existing literature, allowed the discovery of 208 small bioactive peptides, whereas for medium-sized peptides, only one was reported being bioactive.

THE ROLES OF ENZYME IN FOOD PROCESSING - AN OVERVIEW

Enzymes haven used long in food processing before their discovery as a biological catalyst. Food fermentation was among the early art of food processing and the use of enzymes in fermentation and cheese making started about 6000 BC. The roles of enzymes in food processing and preservation contributed to the development of mankind. They contributed in the areas of baking, cheese making, dairy processing, milling, cereals technology, juice and beverages processing, vegetable processing, oils and fats processing, and wine processing among others. Microorganisms are the earliest and foremost source of enzymes used in food processing, other sources are plant and animal tissues and organs. Advances in science and technology disclosed more potentials of enzymes and biotechnology open doors for commercial production of enzymes with charming properties. The development of enzyme immobilization techniques allows the reused of enzymes without affecting their properties, structure, or activities. Recent advances in genetic engineering and recombinant DNA technology permit the production of enzymes with exceptional properties. The current trends in the production of Extremozymes will open doors for using enzymes under extreme conditions of temperature, pH, and pressure. In food, processing enzymes can be used as ingredients, processing aid, or as a catalyst for both pre-and post-consumption catalysis. Enzymes improve the quality, shelf life, stability, and sensory properties of foods. They play important roles in food processing by lowering energy consumption, minimizing waste, producing desired products specifically required, and making foods more affordable, palatable, and available

Summary of “Small Research-Discussion” on Cereal Grains in Japan “Use of Enzymes in Food Processing”

Summary of “Small Research-Discussion” on Cereal Grains in Japan “Use of Enzymes in Food Processing”

Kinetics of growth and co-production of amylase and protease in novel marine actinomycete, Streptomyces lopnurensis KaM5.

Amylases and proteases are among the industrially most important enzymes for food processing, animal feed, brewing, starch processing, detergents, healthcare, leather processing, and biofuel production. In this study, we investigated the growth kinetics and statistically optimized the co-production of amylase and protease in a phylogenetically novel haloalkaliphilic actinomycete, Streptomyces lopnurensis KaM5 of seawater. The Plackett-Berman design using Minitab 14.0 software was employed to assess the impact of the nutritional factors, temperature, pH, and incubation time. Further, starch, yeast extract, NaCl concentrations, and incubation time were optimized by Box-Behnken design at their three levels. The Pareto charts, contour, surface plots, and individual factorial analysis expressed the variability and levels for the optimal enzyme production. ANOVA analysis admitted the statistical fitness and significance level among the variables. A two-fold increase in enzyme production was achieved by cost-effective co-production media. The study was further extended to growth kinetics associated with enzyme production. Specific growth rate (μ), maximal cell mass (Xmax), volumetric product formation (Pmax), rate of product formation (Qp), and generation time (g) were computed and analyzed. These parameters significantly improved when compared with the pre-optimized conditions, and the production economics of the enzyme was industrially viable. The initial studies on the characteristics of the enzymes suggested its ability to function under the combination of alkaline pH and high salt concentrations. The co-production of enzymes from extremophiles can be a potentially viable option for large-scale production and applications.

Enzymes in Food and Beverage Processing

No Abstract.

Extremozymes from metagenome: Potential applications in food processing

ABSTRACTThe long-established use of enzymes for food processing and product formulation has resulted in an increased enzyme market compounding to 7.0% annual growth rate. Advancements in molecular biology and recognition that enzymes with specific properties have application for industrial production of infant, baby and functional foods boosted research toward sourcing the genes of microorganisms for enzymes with distinctive properties. In this regard, functional metagenomics for extremozymes has gained attention on the premise that such enzymes can catalyze specific reactions. Hence, metagenomics that can isolate functional genes of unculturable extremophilic microorganisms has expanded attention as a promising tool. Developments in this field of research in relation to food sector are reviewed.

Production of fungal pectinase enzyme through solid substrate fermentation of estate waste for improvement of enzymatic oxidation and increasing the quality of CTC tea

Enzymes, especially pectinase, plays an important role in the processing of tea that determines the quality and the color appearance of black tea. The main obstacle encountered in the application of enzymes in food processing in general, and in particular in the processing of black tea is the price of commercial enzyme that are still very expen­sive, because it is still imported. Indonesia has a high fungal diversitythat is potential as producer of commercial pectinase enzyme. To reduce the cost of production, fermentation was done using solid waste of plantation that rich of pectin content such as cocoa pod husk that has not been optimally utilized. The research aimed to develop the production technology for pectinase enzymes of fungi through solid substrate fermentation for applications in the processing of CTC black tea. Research includes the isolation and selection of local superior fungi producing enzymes pectinase, optimiza­tion of enzyme production by fungi isolates were selected through a solid substrate fermen­tation using cocoa pod husk waste, production optimization included optimization of fermen­tation time and substrate composition, enzyme extraction of fermented substrate, and appli­cation of pectinase enzyme extracts in the processing of CTC black tea and sensory test of fermented tea. The results showed that it has been obtained isolates of Aspergillus niger on Pectinase Screening Agar Medium (PSAM) which was capable of producing pectinase enzyme on growing media of mix cocoa pod husk and rice bran with additional of enrichment media. The optimum conditions of pectinase production was obtained in the composition of cacao pod husk: rice bran (80:20 w/w) plus enrichment media, the incubation time of 9 days, which produced pectinase activity of 125 U/mL. Applications of pectinase on enzymatic oxidation of CTC tea proved that it was capable of raising the quality of the tea flavor, though the tea gradedid not change.

The use of enzymes in food processing: A review

The use of enzymes in food processing: A review

Diversity of glycosyl hydrolase enzymes from metagenome and their application in food industry.

Traditional use of enzymes for food processing and production of food ingredients resulted in fast-growing enzyme industries world over. The advances in technologies gave rise to exploring newer enzymes and/or modified enzymes for specific application. Search for novel enzymes that can augment catalytic efficiency and advances in molecular biology techniques including sequencing has targeted microbial diversity through metagenomic approaches for sourcing enzymes from difficult to culture organisms. Such mining studies have received more attention in characterizing hydrolases, their prevalence, broad substrate specificities, stability, and independence of cofactors. The focus on glycosyl hydrolases from metagenome for their application in food sector is reviewed.

Displaying Candida antarctica lipase B on the cell surface of Aspergillus niger as a potential food-grade whole-cell catalyst

Aspergillus niger is a recognized workhorse used to produce food processing enzymes because of its extraordinarily high protein-producing capacity. We have developed a new cell surface display system de novo in A. niger using expression elements from generally recognized as safe certified microorganisms. Candida antarctica lipase B (CALB), a widely used hydrolase, was fused to an endogenous cell wall mannoprotein, CwpA, and functionally displayed on the cell surface. Localization of CALB was confirmed by enzymatic assay and immunofluorescence analysis using laser scanning confocal microscopy. After induction by maltose for 45h, the hydrolytic activity and synthesis activity of A. niger mycelium-surface displayed CALB (AN-CALB) reached 400 and 240U/g dry cell, respectively. AN-CALB was successfully used as a whole-cell catalyst for the enzymatic production of ethyl esters from a series of fatty acids of different chain lengths and ethanol. In a solvent-free system, AN-CALB showed great synthetic activity and afforded high substrate mole conversions, which amounted to 87% for ethyl hexanoate after 2h, 89% for ethyl laurate after 2h, and 84% for ethyl stearate after 3h. These results suggested that CwpA can act as an efficient anchoring motif for displaying enzyme on A. niger, and AN-CALB is a robust, green, and cost-effective alternative food-grade whole-cell catalyst to commercial lipase.

The potential for upgrading traditional fermented foods through biotechnology

Fermented foods play an important socio–economic role in developing countries as well as making a major contribution to the protein requirements of natural populations. In general, traditional fermented foods are made under primitive conditions, which result in low yield and poor quality. This paper outlines the present status of some indigenous fermented foods and beverages with some information on the microbiology and biochemistry of the fermentations. Among these are ogi, a fermented cereal gruel used as a weaning food, pito and burukutu, alcoholic cereal beverages. Dawadawa from fermented oil seeds is also popular as nutritious non–meat protein foods, while serving as a condiments and flavors in soups. Traditionally, women carry out fermented food processing activities. The production is craft-based despite the dawn of science and technology. The techniques they use are labour intensive, time consuming and have low productivities, with success depending upon observation of good manufacturing practice. Factors contributing to lack of consumer appeal of indigenous manufactured foods will be also considered. In the change from craft to a technology based production system, several strategies should be adopted. These include the use of starter cultures, stabilization of spontaneous fermentations, and production of food processing enzymes. In view of the considerable range of technologies for improving traditional bioprocessing, the challenges and potential application of biotechnology in upgrading these foods will be discussed. African Journal of Biotechnology Vol. 4 (5), pp. 375-380, 2005

“Recombinant protein of the day”: Using daily student presentations to add real‐world aspects to a biotechnology course

“Recombinant protein of the day”: Using daily student presentations to add real‐world aspects to a biotechnology course

Fungiculture or Termite Husbandry? The Ruminant Hypothesis.

We present a new perspective for the role of Termitomyces fungi in the mutualism with fungus-growing termites. According to the predominant view, this mutualism is as an example of agriculture with termites as farmers of a domesticated fungus crop, which is used for degradation of plant-material and production of fungal biomass. However, a detailed study of the literature indicates that the termites might as well be envisioned as domesticates of the fungus. According to the “ruminant hypothesis” proposed here, termite workers, by consuming asexual fruiting bodies not only harvest asexual spores, but also lignocellulolytic enzymes, which they mix with foraged plant material and enzymes of termite and possibly bacterial origin. This mixture is the building material of the fungus garden and facilitates efficient degradation of plant material. The fungus garden thus functions as an external rumen for termites and primarily the fungi themselves benefit from their own, and gut-derived, lignocellulolytic enzymes, using the termites to efficiently mix these with their growth substrate. Only secondarily the termites benefit, when they consume the degraded, nitrogen-enriched plant-fungus mixture a second time. We propose that the details of substrate use, and the degree of complementarity and redundancy among enzymes in food processing, determine selection of horizontally transmitted fungal symbionts at the start of a colony: by testing spores on a specific, mechanically and enzymatically pre-treated growth substrate, the termite host has the opportunity to select specific fungal symbionts. Potentially, the gut-microbiota thus influence host-fungus specificity, and the selection of specific fungal strains at the start of a new colony. We argue that we need to expand the current bipartite insect-biased view of the mutualism of fungus-growing termites and include the possible role of bacteria and the benefit for the fungi to fully understand the division of labor among partners in substrate degradation.

Genomic Analysis of Koji Mold Aspergillus oryzae and Investigation of Novel Peptidases by Post-genomic Approach

Aspergillus oryzae is a filamentous fungus used as a starter microorganism in the process of manufacturing various kinds of traditional fermented food. It is also used as a producer of enzymes for both food processing and pharmaceuticals, such as proteases and taka-amylase. In order to clarify the function of this microorganism in fermentation industries and utilize the new function of A. oryzae, it was subject to genomic analysis. During the analysis, the telomere of A. oryzae exhibited a novel repeated sequence distinct from other fungi. Genomic analysis also revealed that the genome of A. oryzae consists of eight chromosomes containing 12,074 genes and is larger in size than Aspergillus nidulans and Aspergillus fumigatus. A. oryzae contains sequences specific for A. oryzae for genes involved in primary and secondary metabolism, when compared with A. nidulans and A. fumigatus. Among them, the function of one of the peptidase-like gene products was investigated with a post-genomic approach, and determined as an aminopeptidase specific for acidic amino acids (aspartate and glutamate) which was first identified in filamentous fungi. It also demonstrated that the addition of cobalt ion to the growth medium greatly improved the specific activity. Using the same approach, some of the novel aminopeptidases and carboxypeptidases of A. oryzae were identified and characterized.

Microbial commercial enzymes

This review paper provides a nice overview of developments in the use of enzymes for pulp bleaching. This database provides a tool for finding homologues to commercially useful enzymes by scanning metagenomic sequences and microbial genomes. BRENDA provides an excellent compendium of information on a wide range of enzymes, including microbial enzymes that are useful in biotechnology. Many microbial enzymes that are active with carbohydrate substrates are useful in biotechnology. This database focuses on this class of enzymes. Lipases are heavily used in industry, especially for chiral resolution to produce compounds enantiospecifically. This database deals specifically with lipases. Cytochrome P450 enzymes are widespread in biological systems, including bacteria and fungi. These monooxygenases catalyse important biotransformations, for example with steroid substrates. This website provides a number of very useful tool for people studying serine proteases. Cellulases are now being heavily studied for the production of biofuels starting with cellulosic biomass. The term cellulase actually refers to a class of enzymes that act in concert to hydrolyse biomass cellulose. Verenium has discovered or developed many novel microbial enzymes designed to solve problems in industrial catalysis. This association page provides many links to information on commercial aspects of enzymes. This Wikipedia entry on enzymes has a nice section on enzyme applications. It lists many enzyme applications in a well-organized table. This database provides information of the reaction mechanisms of many important enzymes. The information is well curated with mechanisms broken down into a series of intermediate steps that can be viewed as graphical depictions, each on separate pages. This page provides a nice historical perspective on enzymes and their applications. The Biotechnology Industry Organization provides this list of enzymes, their sources and their applications in the biotechnology industries. This page gives a rosy forecast for the growth of the enzyme business in the near future. Genencor is a major enzyme producer. They express, purify and sell largely microbial enzymes for a broad range of industrial and consumer applications. Nozozymes is the world's largest enzyme-producing company. They make enzymes for biofuels, agriculture, food processing and other applications. The Biocataysis/Biodegradation database has information on many enzymes useful in biotechnology; for example: nitrilases, esterases, epoxide hydrolases, amidases and dehalogenases. This Wikipedia entry highlights the restriction enzymes. Restriction enzymes are produced by bacteria and have found great usefulness in recombinant DNA technology. This page of the EBI IntEnz database highlights the laccase enzymes. Laccases are produced by bacteria and fungi. They have usefulness in the pulp and paper industries and in food processing.

Proteomics of industrial fungi: trends and insights for biotechnology

Filamentous fungi are widely known for their industrial applications, namely, the production of food-processing enzymes and metabolites such as antibiotics and organic acids. In the past decade, the full genome sequencing of filamentous fungi increased the potential to predict encoded proteins enormously, namely, hydrolytic enzymes or proteins involved in the biosynthesis of metabolites of interest. The integration of genome sequence information with possible phenotypes requires, however, the knowledge of all the proteins in the cell in a system-wise manner, given by proteomics. This review summarises the progress of proteomics and its importance for the study of biotechnological processes in filamentous fungi. A major step forward in proteomics was to couple protein separation with high-resolution mass spectrometry, allowing accurate protein quantification. Despite the fact that most fungal proteomic studies have been focused on proteins from mycelial extracts, many proteins are related to processes which are compartmentalised in the fungal cell, e.g. β-lactam antibiotic production in the microbody. For the study of such processes, a targeted approach is required, e.g. by organelle proteomics. Typical workflows for sample preparation in fungal organelle proteomics are discussed, including homogenisation and sub-cellular fractionation. Finally, examples are presented of fungal organelle proteomic studies, which have enlarged the knowledge on areas of interest to biotechnology, such as protein secretion, energy production or antibiotic biosynthesis.

Enzymes in food processing: a condensed overview on strategies for better biocatalysts.

Food and feed is possibly the area where processing anchored in biological agents has the deepest roots. Despite this, process improvement or design and implementation of novel approaches has been consistently performed, and more so in recent years, where significant advances in enzyme engineering and biocatalyst design have fastened the pace of such developments. This paper aims to provide an updated and succinct overview on the applications of enzymes in the food sector, and of progresses made, namely, within the scope of tapping for more efficient biocatalysts, through screening, structural modification, and immobilization of enzymes. Targeted improvements aim at enzymes with enhanced thermal and operational stability, improved specific activity, modification of pH-activity profiles, and increased product specificity, among others. This has been mostly achieved through protein engineering and enzyme immobilization, along with improvements in screening. The latter has been considerably improved due to the implementation of high-throughput techniques, and due to developments in protein expression and microbial cell culture. Expanding screening to relatively unexplored environments (marine, temperature extreme environments) has also contributed to the identification and development of more efficient biocatalysts. Technological aspects are considered, but economic aspects are also briefly addressed.

Determining the safety of enzymes used in animal feed

The purpose of this paper is to provide guidance for evaluating the safety of enzyme preparations used in animal feed. Feed enzymes are typically added to animal feed to increase nutrient bioavailability by acting on feed components prior to or after consumption, i.e., within the gastrointestinal tract. In contrast, food processing enzymes are generally used during processing and then inactivated or removed prior to consumption. The enzymes used in both applications are almost always impure mixtures of active enzyme and other metabolites from the production strain, hence similar safety evaluation procedures for both are warranted. We propose that the primary consideration should be the safety of the production strain and that the decision tree mechanism developed previously for food processing enzymes (Pariza and Johnson, 2001) is appropriate for determining the safety of feed enzymes. Thoroughly characterized non-pathogenic, non-toxigenic microbial strains with a history of safe use in enzyme manufacture are also logical candidates for generating safe strain lineages, from which additional strains may be derived via genetic modification by traditional and non-traditional strategies. For new feed enzyme products derived from a safe strain lineage, it is important to ensure a sufficiently high safety margin for the intended use, and that the product complies with appropriate specifications for chemical and microbial contamination.