Rita P. Lopes

Microorganisms under high pressure — Adaptation, growth and biotechnological potential

Hydrostatic pressure is a well-known physical parameter which is now considered an important variable of life, since organisms have the ability to adapt to pressure changes, by the development of resistance against this variable. In the past decades a huge interest in high hydrostatic pressure (HHP) technology is increasingly emerging among food and biosciences researchers. Microbial specific stress responses to HHP are currently being investigated, through the evaluation of pressure effects on biomolecules, cell structure, metabolic behavior, growth and viability. The knowledge development in this field allows a better comprehension of pressure resistance mechanisms acquired at sub-lethal pressures. In addition, new applications of HHP could arise from these studies, particularly in what concerns to biotechnology. For instance, the modulation of microbial metabolic pathways, as a response to different pressure conditions, may lead to the production of novel compounds with potential biotechnological and industrial applications. Considering pressure as an extreme life condition, this review intends to present the main findings so far reported in the scientific literature, focusing on microorganisms with the ability to withstand and to grow in high pressure conditions, whether they have innated or acquired resistance, and show the potential of the application of HHP technology for microbial biotechnology.

Hyperbaric storage of melon juice at and above room temperature and comparison with storage at atmospheric pressure and refrigeration

Hyperbaric storage (8h) of melon juice (a highly perishable food) at 25, 30 and 37°C, under pressure at 25-150 MPa was compared with atmospheric pressure storage (0.1 MPa) at the same temperatures and under refrigeration (4°C). Comparatively to the refrigerated condition, hyperbaric storage at 50/75 MPa resulted in similar or lower microbial counts (total aerobic mesophiles, enterobacteriaceae, and yeasts/moulds) while at 100/150 MPa, the counts were lower for all the tested temperatures, indicating in the latter case, in addition to microbial growth inhibition, a microbial inactivation effect. At 25 MPa no microbial inhibition was observed. Physicochemical parameters of all samples stored under pressure (pH, titratable acidity, total soluble solids, browning degree and cloudiness) did not show a clear variation trend with pressure, being the results globally similar to refrigeration storage. These results show the potential of hyperbaric storage, at and above room temperature and with potential energy savings, comparatively to refrigeration.

Fermentation at non-conventional conditions in food- and bio-sciences by the application of advanced processing technologies

Abstract The interest in improving the yield and productivity values of relevant microbial fermentations is an increasingly important issue for the scientific community. Therefore, several strategies have been tested for the stimulation of microbial growth and manipulation of their metabolic behavior. One promising approach involves the performance of fermentative processes during non-conventional conditions, which includes high pressure (HP), electric fields (EF) and ultrasound (US). These advanced technologies are usually applied for microbial inactivation in the context of food processing. However, the approach described in this study focuses on the use of these technologies at sub-lethal levels, since the aim is microbial growth and fermentation under these stress conditions. During these sub-lethal conditions, microbial strains develop specific genetic, physiologic and metabolic stress responses, possibly leading to fermentation products and processes with novel characteristics. In some cases, these modifications can represent considerable improvements, such as increased yields, productivities and fermentation rates, lower accumulation of by-products and/or production of different compounds. Although several studies report the successful application of these technologies during the fermentation processes, information on this subject is still scarce and poorly understood. For that reason, the present review paper intends to assemble and discuss the main findings reported in the literature to date, and aims to stimulate interest and encourage further developments in this field.

Implementation of Emerging Technologies

Novel processing technologies have been gaining interest among food researchers due to their lower impact on nutritional and sensory properties of the products compared to the conventional thermal techniques. In this chapter some of the most well-studied (eg, high-pressure processing, pulsed electric fields, ohmic heating, microwave, and ultrasound) emerging technologies are briefly reviewed. Most of these technologies have found niche applications in the food industry, replacing or complementing conventional preservation technologies. Thereby, data on commercialization, energy, and microbial safety are presented and discussed with an ultimate goal to explore strategies for their implementation in the food industry. Novel thermal and nonthermal technologies have shown clear environmental benefits by improving the overall energy efficiency of the process and reducing the use of nonrenewable resources. Lastly, studies with specific examples of the implementation of these novel processing technologies in food industry are described, focusing on the application of high-pressure processing and pulsed electric fields to orange juice, milk, and oysters. Higher implementation costs were observed for the emerging processing technologies compared to the conventional processing technologies, which can be explained by the fact that the industrial application of these novel technologies is still under development. In the future the costs are expected to reduce with further technology advances and their increasing implementation in the industry.

Application of High Pressure with Homogenization, Temperature, Carbon Dioxide, and Cold Plasma for the Inactivation of Bacterial Spores: A Review

Formation of highly resistant spores is a concern for the safety of low-acid foods as they are a perfect vehicle for food spoilage and/or human infection. For spore inactivation, the strategy usually applied in the food industry is the intensification of traditional preservation methods to sterilization levels, which is often accompanied by decreases of nutritional and sensory properties. In order to overcome these unwanted side effects in food products, novel and emerging sterilization technologies are being developed, such as pressure-assisted thermal sterilization, high-pressure carbon dioxide, high-pressure homogenization, and cold plasma. In this review, the application of these emergent technologies is discussed, in order to understand the effects on bacterial spores and their inactivation and thus ensure food safety of low-acid foods. In general, the application of these novel technologies for inactivating spores is showing promising results. However, it is important to note that each technique has specific features that can be more suitable for a particular type of product. Thus, the most appropriate sterilization method for each product (and target microorganisms) should be assessed and carefully selected.

Growth and metabolism of Oenococcus oeni for malolactic fermentation under pressure.

Malolactic fermentation is a biological deacidification process of wine, characterized by the transformation of l‐malic acid to l‐lactic acid and CO2. Oenococcus oeni is able to perform malolactic fermentation and to survive under wine harsh conditions, representing great interest for wine industry. The aim of this work was to evaluate the effect of high pressure on the metabolism of O. oeni growing in culture media, regarding malolactic fermentation, sugars metabolism and bacterial growth. A pressure stress of 50 MPa during 8 h did not result in significant modifications in bacterial metabolism. In contrast, a stress of 100 MPa during 8 h resulted in lower amounts of l‐lactic acid, while higher amounts of d‐lactic acid were also registered, indicating changes in bacterial metabolism. A pressure stress of 0·5 MPa during 300 h resulted in complete inactivation of O. oeni, but malolactic fermentation was still observed at some extent, showing that malolactic enzyme was not completely inactivated at these conditions. It was concluded that high pressure causes modification of O. oeni metabolism, and possibly in enzyme activities.

Lactobacillus reuteri growth and fermentation under high pressure towards the production of 1,3-propanediol

Lactobacillus reuteri is a lactic acid bacterium able to produce several relevant bio-based compounds, including 1,3-propanediol (1,3-PDO), a compound used in food industry for a wide range of purposes. The performance of fermentations under high pressure (HP) is a novel strategy for stimulation of microbial growth and possible improvement of fermentation processes. Therefore, the present work intended to evaluate the effects of HP (10-35 MPa) on L. reuteri growth and glycerol/glucose co-fermentation, particularly on 1,3-PDO production. Two different types of samples were used: with or without acetate added in the culture medium. The production of 1,3-PDO was stimulated at 10 MPa, resulting in enhanced final titers, yields and productivities, compared to 0.1 MPa. The highest 1,3-PDO titer (4.21 g L-1) was obtained in the presence of acetate at 10 MPa, representing yield and productivity improvements of ≈ 11 and 12%, respectively, relatively to the same samples at 0.1 MPa. In the absence of acetate, 1,3-PDO titer and productivity were similar to 0.1 MPa, but the yield increased ≈ 26%. High pressure also affected the formation of by-products (lactate, acetate and ethanol) and, as a consequence, higher molar ratios 1,3-PDO:by-products were achieved at 10 MPa, regardless of the presence/absence of acetate. This indicates a metabolic shift, with modification of product selectivity towards production of 1,3-PDO. Overall, this work suggests that HP can be a useful tool to improve of 1,3-PDO production from glycerol by L. reuteri, even if proper process optimization and scale-up are still needed to allow its industrial application. It also opens the possibility of using this technology to stimulate other glycerol fermentations processes that are relevant for food science and biotechnology.

Pasteurization: Effect on Sensory Quality and Nutrient Composition

Thermal pasteurization is a mild heat treatment commonly used for the preservation of a wide variety of foods. However, it may have negative effects on food quality, and, consequently, novel pasteurization technologies have emerged: high pressure processing, pulsed electric fields, ohmic heating, microwave, and others. These nonconventional techniques may also affect food properties, depending on food matrix and treatment conditions. Further work is needed to optimize the process conditions that will guarantee food safety while minimizing its impact on food quality. This article summarizes the knowledge available concerning the effect of different pasteurization technologies on nutritional and sensorial aspects of foods.