Development of Wide-Spectrum Hybrid Bacteriocins for Food Biopreservation

The food industry demands new procedures and methods to produce minimally processed, ready to eat food with intact nutritional, taste, and flavor properties. The biopreservation and the use of both bacteriocins produced by lactic acid bacteria (LAB) and bacteriocinogenic strains as an alternative to substitute chemical antimicrobial for food preservation became increasingly important in the last two decades. When the new proposed natural preservatives techniques are applied, probiotics food can be obtained and, simultaneously, foodborne pathogens and spoilage contaminants can diminish. However, bacteriocins produced by LAB have a narrow antibacterial spectrum and are inactive against Gram-negative bacteria like Salmonella and the emergent enterohemorrhagic Escherichia coli. Knowing the mechanism of action and the structural features of microcins synthesized by Gram-negative bacteria and with potent antimicrobial activity against the mentioned microorganism, the proposal is to obtain hybrid peptides (microcin–bacteriocin) with broad antimicrobial spectrum. This review explains how the inability of bacteriocins to cross the outer membrane of Gram-negative bacteria unable them to act on the bacteria. It will also be discussed how a hybrid bacteriocin can be obtained.     

Bacteriocin-based strategies for food biopreservation

Bacteriocins are ribosomally-synthesized peptides or proteins with antimicrobial activity, produced by different groups of bacteria. Many lactic acid bacteria (LAB) produce bacteriocins with rather broad spectra of inhibition. Several LAB bacteriocins offer potential applications in food preservation, and the use of bacteriocins in the food industry can help to reduce the addition of chemical preservatives as well as the intensity of heat treatments, resulting in foods which are more naturally preserved and richer in organoleptic and nutritional properties. This can be an alternative to satisfy the increasing consumers demands for safe, fresh-tasting, ready-to-eat, minimally-processed foods and also to develop "novel" food products (e.g. less acidic, or with a lower salt content). In addition to the available commercial preparations of nisin and pediocin PA-1/AcH, other bacteriocins (like for example lacticin 3147, enterocin AS-48 or variacin) also offer promising perspectives. Broad-spectrum bacteriocins present potential wider uses, while narrow-spectrum bacteriocins can be used more specifically to selectively inhibit certain high-risk bacteria in foods like Listeria monocytogenes without affecting harmless microbiota. Bacteriocins can be added to foods in the form of concentrated preparations as food preservatives, shelf-life extenders, additives or ingredients, or they can be produced in situ by bacteriocinogenic starters, adjunct or protective cultures. Immobilized bacteriocins can also find application for development of bioactive food packaging. In recent years, application of bacteriocins as part of hurdle technology has gained great attention. Several bacteriocins show additive or synergistic effects when used in combination with other antimicrobial agents, including chemical preservatives, natural phenolic compounds, as well as other antimicrobial proteins. This, as well as the combined use of different bacteriocins may also be an attractive approach to avoid development of resistant strains. The combination of bacteriocins and physical treatments like high pressure processing or pulsed electric fields also offer good opportunities for more effective preservation of foods, providing an additional barrier to more refractile forms like bacterial endospores as well. The effectiveness of bacteriocins is often dictated by environmental factors like pH, temperature, food composition and structure, as well as the food microbiota. Foods must be considered as complex ecosystems in which microbial interactions may have a great influence on the microbial balance and proliferation of beneficial or harmful bacteria. Recent developments in molecular microbial ecology can help to better understand the global effects of bacteriocins in food ecosystems, and the study of bacterial genomes may reveal new sources of bacteriocins.     

Combating biofilms of foodborne pathogens with bacteriocins by lactic acid bacteria in the food industry

Most foodborne pathogens have biofilm-forming capacity and prefer to grow in the form of biofilms. Presence of biofilms on food contact surfaces can lead to persistence of pathogens and the recurrent cross-contamination of food products, resulting in serious problems associated with food safety and economic losses. Resistance of biofilm cells to conventional sanitizers urges the development of natural alternatives to effectively inhibit biofilm formation and eradicate preformed biofilms. Lactic acid bacteria (LAB) produce bacteriocins which are ribosomally synthesized antimicrobial peptides, providing a great source of nature antimicrobials with the advantages of green and safe properties. Studies on biofilm control by newly identified bacteriocins are increasing, targeting primarily onListeria monocytogenes, Staphylococcus aureus, Salmonella, and Escherichia coli. This review systematically complies and assesses the antibiofilm property of LAB bacteriocins in controlling foodborne bacterial-biofilms on food contact surfaces. The bacteriocin-producing LAB genera/species, test method (inhibition and eradication), activity spectrum and surfaces are discussed, and the antibiofilm mechanisms are also argued. The findings indicate that bacteriocins can effectively inhibit biofilm formation in a dose-dependent manner, but are difficult to disrupt preformed biofilms. Synergistic combination with other antimicrobials, incorporation in nanoconjugates and implementation of bioengineering can help to strengthen their antibiofilm activity. This review provides an overview of the potential and application of LAB bacteriocins in combating bacterial biofilms in food processing environments, assisting in the development and widespread use of bacteriocin as a promising antibiofilm-agent in food industries.     

乳酸菌细菌素抗菌潜力挖掘研究进展

乳酸菌产生的主要抗菌防腐物质是有机酸,但在乳酸菌代谢产物中发现了细菌素等其它抗菌物质,其中有的细菌素甚至具有强烈的抗真菌能力。世界范围内启动的乳酸菌基因组计划创造了巨大的生物信息流。乳酸菌基因组蕴含的生物信息将对研究乳酸菌细菌素的合成代谢途径,功能基因挖掘以及新型抗菌剂的开发搭建信息平台。结合乳酸菌细菌素的最新研究进展,讨论利用乳酸菌基因组探索乳酸菌的抗菌潜力及其作为生物防腐剂在食品保藏中的应用前景。     

Enzymatic Production,Bioactivity, and Bitterness of Chickpea (Cicer arietinum) Peptides

Chickpeas are inexpensive, protein rich (approximately 20% dry mass) pulses available worldwide whose consumption has been correlated with positive health outcomes. Dietary peptides are important molecules derived from dietary proteins, but a comprehensive analysis of the peptides that can be produced from chickpea proteins is missing in the literature. This review provides information from the past 20 years on the enzymatic production of peptides from chickpea proteins, the reported bioactivities of chickpea protein hydrolysates and peptides, and the potential bitterness of chickpea peptides in food products. Chickpea peptides have been enzymatically produced with pepsin, trypsin, chymotrypsin, alcalase, flavorzyme, and papain either alone or in combination, but the sequences of many of the peptides in chickpea protein hydrolysates remain unknown. In addition, a theoretical hydrolysis of chickpea legumin by stem bromelain and ficin was performed by the authors to highlight the potential use of these enzymes to produce bioactive chickpea peptides. Antioxidant activity, hypocholesterolemic, and angiotensin 1‐converting enzyme inhibition are the most studied bioactivities of chickpea protein hydrolysates and peptides, but anticarcinogenic, antimicrobial, and anti‐inflammatory effects have also been reported for chickpea protein hydrolysates and peptides. Chickpea bioactive peptides are not currently commercialized, but their bitterness could be a major impediment to their incorporation in food products. Use of flavorzyme in the production of chickpea protein hydrolysates has been proposed to decrease their bitterness. Future research should focus on the optimization of chickpea bioactive peptide enzymatic production, studying the bioactivity of chickpea peptides in humans, and systematically analyzing chickpea peptide bitterness.     

Overview of Lactobacillus plantarum as a promising bacteriocin producer among lactic acid bacteria

Chemical preservatives have been traditionally used during the manufacturing of processed products. However, the continuous growing interest of consumers for fresh and natural products makes it necessary to search for alternative compounds. In this context, food industries have been widely using lactic acid bacteria (LAB) as natural preservatives, due to their ability to produce antibacterial compounds such as bacteriocins. Similarly, pharmaceutical industries have improved the use of these bacterial peptides, with antibacterial activity, trying to reduce the indiscriminate use of antibiotics in food products for human and animal consumption. Among LAB, Lactobacillus plantarum can be adapted to various niches thanks to its ability to ferment a wide range of carbohydrates. Additionally, it can be used as starter culture in food fermentations and as an ingredient for probiotic foods, contributing to the organoleptic characteristics of foods at the same time prolonging the shelf-life and safety of these products. The amount of valuable substances obtained from L. plantarum species isolated from different ecological niches is also worth noting, thus proving it to be one of the most important and versatile species among LAB.     

提高乳酸菌细菌素合成量方法的研究进展

细菌素是某些细菌通过核糖体合成机制产生的蛋白质或多肽,能够抑制与其亲源关系相同或相近的微生物,某些细菌素在食品加工和发酵过程中能抑制致病菌和腐败菌。乳酸菌被认为是一般公认安全,其细菌素具有安全性高、稳定性好、抑菌谱广等优点,作为一种新型食品防腐剂备受关注,但商品化的乳酸菌细菌素十分有限,仅限于Nisin和Pediocin PA-1等少数几种,合成量低是细菌素在食品中应用受限的主要原因之一。从不同原料中筛选高产菌株、发酵培养基和发酵条件优化、诱变育种、原生质体融合、基因工程方法、群体感应系统调控六个方面,论述了增加乳酸菌细菌素合成量的方法,以期为实现乳酸菌细菌素的工业化生产提供一定的借鉴。     

A New Frontier in Soy Bioactive Peptides that May Prevent Age‐related Chronic Diseases

During gastrointestinal digestion or food processing of proteins, small peptides can be released and may act as regulatory compounds with hormone‐like activities. Numerous biologically active peptides (bioactive peptides) have been identified. Most bioactive peptides are derived from milk and dairy products, with the most common being angiotensin converting enzyme inhibitory peptides. Soybean protein and soybean derived peptides also play an important role in soybean physiological activities, particularly those related to the prevention of chronic diseases. However, the bioactive potential of soybean derived bioactive peptides is yet to be fully appreciated. After a general introduction of approaches and advances in bioactive peptides from food sources, this review focuses on bioactive peptides derived from soybean proteins and their physiological properties. Technological approaches to generate bioactive peptides, their isolation, purification, characterization, and quantification, and further application in food and drug design are also presented. Safety concerns, such as potential toxicity, allergenicity, and sensory aspect of these peptides are likewise discussed.     

Pediocin PA-1的结构、生物合成与作用机制

乳酸菌素是乳酸菌通过核糖体合成机制产生的一类具有生物活性的蛋白质、多肽或前体多肽.最近将乳酸菌素分为三大类,其中Ⅱ a类类片球菌素是研究的热点,它的典型代表是Pediocin PA-1.Pedioein PA-1具有较强的抗单增李斯特菌特性和热稳定性,是一种具有广阔应用前景的食品添加剂.本文综述了Pediocin PA-1的结构、生物合成和作用机制,概述了国内外对Pediocin PA-1的研究进展和发展方向,以期对细菌素的深入研究和广泛应用起到抛砖引玉的作用.     

群体感应系统在乳酸菌产细菌素中的作用

许多乳酸菌能够产生抗菌活性肽——细菌素,细菌素具有不同的结构、作用方式、抑菌谱和效价,通常认为乳酸菌和其所产的细菌素都是安全的,乳酸菌所产细菌素作为天然食品防腐剂已显示了巨大的潜能。基于群体感应的细胞间交流已成为细菌素合成的关键调控机制,群体感应作为细胞密度函数,可使细菌素产生保持同步性。群体感应需通过信号分子介导感知菌体密度,信号分子随着菌体密度增加而增加,并激活信号转导级联使菌体产生细菌素。本文通过对乳酸菌群体感应信号分子种类、信号转导机制及群体感应系统对两类细菌素合成的调控进行综述,以初步了解群体感应系统在乳酸菌产细菌素过程中的作用机制。     

Preservation and fermentation: past,present and future

Preservation of food and beverages resulting from fermentation has been an effective form of extending the shelf-life of foods for millennia. Traditionally, foods were preserved through naturally occurring fermentations, however, modern large scale production generally now exploits the use of defined strain starter systems to ensure consistency and quality in the final product. This review will mainly focus on the use of lactic acid bacteria (LAB) for food improvement, given their extensive application in a wide range of fermented foods. These microorganisms can produce a wide variety of antagonistic primary and secondary metabolites including organic acids, diacetyl, CO2 and even antibiotics such as reuterocyclin produced by Lactobacillus reuteri. In addition, members of the group can also produce a wide range of bacteriocins, some of which have activity against food pathogens such as Listeria monocytogenes and Clostridium botulinum. Indeed, the bacteriocin nisin has been used as an effective biopreservative in some dairy products for decades, while a number of more recently discovered bacteriocins, such as lacticin 3147, demonstrate increasing potential in a number of food applications. Both of these lactococcal bacteriocins belong to the lantibiotic family of posttranslationally modified bacteriocins that contain lanthionine, beta-methyllanthionine and dehydrated amino acids. The exploitation of such naturally produced antagonists holds tremendous potential for extension of shelf-life and improvement of safety of a variety of foods.     

Physical and nutritional conditions for optimized production of bacteriocins by lactic acid bacteria – A review

ABSTRACT

There has been an increasing debate about the use of synthetic chemical compounds and the consequences of their use in food preservation. In this context, the utilization of some natural compounds produced by bacteria, showing an inhibitory effect against microorganisms associated with food contamination, have gained attention as preservation technology. In order to improve the production and yield costs of bacteriocins, detailed studies are necessary to determine the conditions that allow an optimized production and extraction of bacteriocins from lactic acid bacteria (LAB). In this context, this article aims to discuss the information regarding the main factors that influence bacteriocin production by LAB. The biosynthesis of bacteriocins can be influenced by various culture conditions, such as the composition of the medium, pH, temperature and growth kinetics of the microorganisms. One of the limiting factors for the use of bacteriocins on a large scale in food preservation is the economic factor. In order for the production costs of bacteriocins to be reduced, making them attractive, it is necessary to know the optimum parameters of production, thus maximizing productivity and making costs more attractive.     

Heterologous production of bacteriocins by lactic acid bacteria

Over the last two decades, bacteriocins produced by lactic acid bacteria (LAB) have been the subject of considerable research and industrial interest due to their potential as food biopreservatives. The development of heterologous expression systems for such antimicrobial compounds may offer a number of advantages over native systems, such as facilitating the control of bacteriocin gene expression or achieving higher production levels. In addition, the heterologous production by food-grade LAB offers an attractive method for overcoming some of the adverse situations that may affect the effectiveness of some bacteriocins in food systems. Construction of multibacteriocinogenic strains or acquisition of antimicrobial properties by industrial strains are further objectives that can be achieved through the use of heterologous gene expression systems. The development of new biotechnological tools and recent advances in LAB genetics account for the escalating number of studies dealing with heterologous production of bacteriocins by such hosts. This paper reviews the literature published on the subject and compares the different experimental strategies that have been used up to the present for this purpose.     

Possibilities for the utilisation of marine by‐products

Fish industry by‐products can account for up to 75% of the catch depending on postharvest or industrial preparation processes. Different terms such as ‘fish waste’, ‘by‐product’ and ‘rest raw materials’ have been used. The review gives an overview of value‐added processes that provide an alternative to low‐profit uses such as silage, fish meal and mince. The preparation of different by‐product fractions such as fish blood, marine lipids, omega‐3 fatty acids, fish protein fractions and bioactive components with nutraceutical potential, i.e. antioxidants and bioactive peptides, is considered. There are several future opportunities for the preparation of high‐value by‐products such as enzymes, minerals and other bioactive substances including hydroxyapatite, phosphorus, taurine and creatine. Both regulatory status and future market potential need to be considered. In addition, there is a need for technologies that maintain good quality by‐products and ‘simple’ processes to produce bulk products for further refining.     

The importance of lactic acid bacteria for the prevention of bacterial growth and their biogenic amines formation: A review

Foodborne pathogens (FBP) represent an important threat to the consumers' health as they are able to cause different foodborne diseases. In order to eliminate the potential risk of those pathogens, lactic acid bacteria (LAB) have received a great attention in the food biotechnology sector since they play an essential function to prevent bacterial growth and reduce the biogenic amines (BAs) formation. The foodborne illnesses (diarrhea, vomiting, and abdominal pain, etc.) caused by those microbial pathogens is due to various reasons, one of them is related to the decarboxylation of available amino acids that lead to BAs production. The formation of BAs by pathogens in foods can cause the deterioration of their nutritional and sensory qualities. BAs formation can also have toxicological impacts and lead to different types of intoxications. The growth of FBP and their BAs production should be monitored and prevented to avoid such problems. LAB is capable of improving food safety by preventing foods spoilage and extending their shelf-life. LAB are utilized by the food industries to produce fermented products with their antibacterial effects as bio-preservative agents to extent their storage period and preserve their nutritive and gustative characteristics. Besides their contribution to the flavor for fermented foods, LAB secretes various antimicrobial substances including organic acids, hydrogen peroxide, and bacteriocins. Consequently, in this paper, the impact of LAB on the growth of FBP and their BAs formation in food has been reviewed extensively.     

The Evolution and Versatility of Microalgal Biotechnology: A Review

Microalgal biotechnology has emerged due to the health‐promoting properties of microalgae related to their bioactive compounds and the great diversity of products that can be developed from algal biomass. Microalgal biomasses have been produced industrially for applications in different fields such as food, pharmaceutical, nutraceutical, cosmetic, and animal feed industries. They can be cultivated either in open systems or in closed systems (photobioreactors). Another important area is the use of microalgal biomass for energy production. It has become obvious that petroleum‐derived fuels are unsustainable, due to depleting world reserves and greenhouse gas emissions. Microalgae can provide several different types of renewable biofuels. These include methane produced by anaerobic digestion of the algal biomass, biodiesel derived from trans‐esterification of microalgal lipids, bioethanol produced from carbohydrate fermentations, and photobiologically produced biohydrogen. The idea of using microalgae as a source of fuel is not new. However, it is now being taken seriously because of increases in petroleum prices and, more significantly, the increasing concern about global warming as associated with burning fossil fuels. This review offers an update on information about microalgae, specifically emphasizing their biotechnological importance.     

FUNDAMENTALS AND PERSPECTIVES FOR THE USE OF BACTERIOCINS PRODUCED BY LACTIC ACID BACTERIA IN MEAT PRODUCTS

Bacteriocins of lactic acid bacteria (LAB) are proteinaceous compounds that may present antimicrobial activity towards important foodborne pathogens and spoilage-related microflora. Due to these properties, bacteriocin-producing strains or purified bacteriocins have a great potential of use in biologically based food preservation systems. Despite the growing number of articles describing the isolation of bacteriocinogenic strains, genetic determinants for production, as well as the purification and biochemical characterization of these inhibitory substances, there are only limited reports of successful application of bacteriocins to meats.

This paper presents a critical review of the methods available for screening of bacteriocin-producing LAB strains from meats and also discusses the proposed mechanisms of action for LAB bacteriocins. Additionally, an overview of the Brazilian experience in the application of LAB bacteriocins to meats and meat products is given.     

FUNDAMENTALS AND PERSPECTIVES FOR THE USE OF BACTERIOCINS PRODUCED BY LACTIC ACID BACTERIA IN MEAT PRODUCTS

Bacteriocins of lactic acid bacteria (LAB) are proteinaceous compounds that may present antimicrobial activity towards important foodborne pathogens and spoilage-related microflora. Due to these properties, bacteriocin-producing strains or purified bacteriocins have a great potential of use in biologically based food preservation systems. Despite the growing number of articles describing the isolation of bacteriocinogenic strains, genetic determinants for production, as well as the purification and biochemical characterization of these inhibitory substances, there are only limited reports of successful application of bacteriocins to meats.

This paper presents a critical review of the methods available for screening of bacteriocin-producing LAB strains from meats and also discusses the proposed mechanisms of action for LAB bacteriocins. Additionally, an overview of the Brazilian experience in the application of LAB bacteriocins to meats and meat products is given.     

Antihypertensive peptides of animal origin: A review

Many bioactive peptides trigger certain useful antihypertensive activities in the living body system and there is a mounting worldwide interest in the therapeutic potential of these bioactive peptides for exploitation in vivo against the hypertension. Studies suggest the antihypertensive properties for many bioactive peptides of animal origin with underlying mechanisms ranging from inhibition of angiotensin-converting enzyme to additional mechanisms to lower blood pressure such as opioid-like activities and mineral-binding and antithrombotic properties. Antihypertensive peptides are the most extensively studied of all the bioactivities induced by food protein hydrolysates, highlighting their importance in human health and disease prevention and treatment. There exist enormous opportunities for the production of novel peptide-based products in biopharmaceutical manufacturing industries for the treatment, prevention, and mitigation of hypertension. Numerous products have already struck on the global market and many more are in process. This article focuses on antihypertensive peptides identified in the meat, fish, blood, milk, dairy products, and egg and their probable application as novel ingredients in the development of functional food products as dietary treatment of hypertension.