Production of microbial transglutaminase by Streptoverticillium cinnamoneum KKP 1658.

Transglutaminase finds broad applications in the food industry, influencing texture, shelf life and overall food quality. It can be utilized to create products with enhanced sensory and technological properties and serves as a tool to reduce food waste. The aim of this study was to optimize the production of microbial transglutaminase (MTG) by the genetically unmodified strain of Streptoverticillium cinnamoneum KKP 1658. Tryptone soy broth (TSB) was chosen as the optimal inoculation medium due to its high MTG activity in the cultivation substrate. The optimal inoculum incubation time was determined as 24 hours, with a dosage of 10 %. Various nitrogen sources were investigated while maintaining a consistent nitrogen dosage (0.2 %) (including aminobak, corn steep liquor, ammonium nitrate and ammonium sulphate) to achieve the highest microbiological transglutaminase activity. The combination of aminobak with corn steep liquor and a cultivation period of 72 hours (28 °C; pH 6.0-6.5) yielded the highest MTG activity at 6.59 U/mL.

Microbiological transglutaminase: Biotechnological application in the food industry.

Microbial transglutaminases (mTGs) belong to the family of global TGs, isolated and characterised by various bacterial strains, with the first being Streptomyces mobaraensis. This literature review also discusses TGs of animal and plant origin. TGs catalyse the formation of an isopeptide bond, cross-linking the amino and acyl groups. Due to its broad enzymatic activity, TG is extensively utilised in the food industry. The annual net growth in the utilisation of enzymes in the food processing industry is estimated to be 21.9%. As of 2020, the global food enzymes market was valued at around $2.3 billion USD (mTG market was estimated to be around $200 million USD). Much of this growth is attributed to the applications of mTG, benefiting both producers and consumers. In the food industry, TG enhances gelation and modifies emulsification, foaming, viscosity, and water-holding capacity. Research on TG, mainly mTG, provides increasing insights into the wide range of applications of this enzyme in various industrial sectors and promotes enzymatic processing. This work presents the characteristics of TGs, their properties, and the rationale for their utilisation. The review aims to provide theoretical foundations that will assist researchers worldwide in building a methodological framework and furthering the advancement of biotechnology research.

Recent advances and opportunities related to the use of bee products in food processing.

Nowadays, natural foods that can provide positive health effects are gaining more and more popularity. Bees and the products they produce are our common natural heritage that should be developed. In the article, we presented the characteristics of bee products and their use in industry. We described the development and importance of beekeeping in the modern world. Due to their high nutritional value and therapeutic properties, bee products are of great interest and their consumption is constantly growing. The basis for the use of bee products in human nutrition is their properties and unique chemical composition. The conducted research and opinions confirm the beneficial effect of bee products on health. The current consumer awareness of the positive impact of food having a pro-health effect on health and well-being affects the increase in interest and demand for this type of food among various social groups. Enriching the daily diet with bee products may support the functioning of the organism. New technologies have appeared on the market to improve the process of obtaining bee products. The use of bee products plays a large role in many industries; moreover, the consumption of bee products and promotion of their medicinal properties are very important in shaping proper eating habits.

Reprocessing of side-streams towards obtaining valuable bacterial metabolites.

Every year, all over the world, the industry generates huge amounts of residues. Side-streams are most often used as feed, landfilled, incinerated, or discharged into sewage. These disposal methods are far from perfect. Taking into account the composition of the side-streams, it seems that they should be used as raw materials for further processing, in accordance with the zero-waste policy and sustainable development. The article describes the latest achievements in biotechnology in the context of bacterial reprocessing of residues with the simultaneous acquisition of their metabolites. The article focuses on four metabolites - bacterial cellulose, propionic acid, vitamin B12 and PHAs. Taking into account global trends (e.g. food, packaging, medicine), it seems that in the near future there will be a sharp increase in demand for this type of compounds. In order for their production to be profitable and commercialised, cheap methods of its obtaining must be developed. The article, in addition to obtaining these bacterial metabolites from side-streams, also discusses e.g. factors affecting their production, metabolic pathways and potential and current applications. The presented chapters provide a complete overview of the current knowledge on above metabolites, which can be helpful for the academic and scientific communities and the several industries. KEY POINTS: • The industry generates millions of tons of organic side-streams each year. • Generated residues burden the natural environment. • A good and cost-effective method of side-streams management seems to be biotechnology - reprocessing with the use of bacteria. • Biotechnological disposal of side-streams gives the opportunity to obtain valuable compounds in cheaper ways: BC, PA, vitmain B12, PHAs.

Effect of selenium and methods of protein extraction on the proteomic profile of Saccharomyces yeast.

Selenium may influence the biosynthesis of individual proteins in the yeast cell cytosol. In this study, we used two-dimensional (2D) electrophoresis to identify proteins that are differentially expressed by the enrichment of selenium in Saccharomyces cerevisiae yeast cells. We chose eight protein fractions for further proteomic analysis. A detailed analysis was performed using the Ultraflextreme matrix-assisted laser desorption/ionisation time-of-flight/time-of-flight mass spectrometer, which enables fast and accurate measurement of the molecular weight of the analysed proteins. This study, for the first time, provides evidence that selenium-enriched yeast contains higher levels of mitochondria malate dehydrogenase, adenosine-5'-triphosphate (ATP)-dependent RNA helicase dbp3, and tryptophan dimethylallyltransferase, and alanyl-tRNA editing protein AlaX than yeast without the addition of selenium. It should be emphasised that the proteomic variability obtained reflects the high biological and complexity of yeast metabolism under control and selenium-enriched conditions and can be properly used in the future as a model for further research aimed at determining the expression of appropriate metabolic genes.

Use of apple pomace, glycerine, and potato wastewater for the production of propionic acid and vitamin B12.

Propionic acid bacteria (PAB) are a source of valuable metabolites, including propionic acid and vitamin B12. Propionic acid, a food preservative, is synthesized from petroleum refining by-products, giving rise to ecological concerns. Due to changing food trends, the demand for vitamin B12 has been expected to increase in the future. Therefore, it is necessary to look for new, alternative methods of obtaining these compounds. This study was conducted with an aim of optimizing the production of PAB metabolites using only residues (apple pomace, waste glycerine, and potato wastewater), without any enzymatic or chemical pretreatment and enrichment. Media consisting of one, two, or three industrial side-streams were used for the production of PAB metabolites. The highest production of propionic acid was observed in the medium containing all three residues (8.15 g/L, yield: 0.48 g/g). In the same medium, the highest production of acetic acid was found - 2.31 g/L (0.13 g/g). The presence of waste glycerine in the media had a positive effect on the efficiency of propionic acid production and P/A ratio. The concentration of vitamin B12 obtained in the wet biomass of Propionibacterium freudenreichii DSM 20271 ranged from 90 to 290 µg/100 g. The highest production of cobalamin was achieved in potato wastewater and apple pomace, which may be a source of the precursors of vitamin B12 - cobalt and riboflavin. The results obtained show both propionic acid and vitamin B12 can be produced in a more sustainable manner through the fermentation of residues which are often not properly managed. KEY POINTS: • The tested strain has been showed metabolic activity in the analyzed industrial side-streams (apple pomace, waste glycerine, potato wastewater). • All the side-streams were relevant for the production of propinic acid. • The addition of waste glycerine increases the propionic acid production efficiency and P/A ratio. • B12 was produced the most in the media containing potato wastewater and apple pomace as dominant ingredients.

Use of Propionibacterium freudenreichii T82 Strain for Effective Biosynthesis of Propionic Acid and Trehalose in a Medium with Apple Pomace Extract and Potato Wastewater.

Propionic acid bacteria are the source of many metabolites, e.g., propionic acid and trehalose. Compared to microbiological synthesis, the production of these metabolites by petrochemical means or enzymatic conversion is more profitable. The components of microbiological media account for a large part of the costs associated with propionic fermentation, due to the high nutritional requirements of Propionibacterium. This problem can be overcome by formulating a medium based on the by-products of technological processes, which can act as nutritional sources and at the same time replace expensive laboratory preparations (e.g., peptone and yeast extract). The metabolic activity of P. freudenreichii was investigated in two different breeding environments: in a medium containing peptone, yeast extract, and biotin, and in a waste-based medium consisting of only apple pomace and potato wastewater. The highest production of propionic acid amounting to 14.54 g/L was obtained in the medium containing apple pomace and pure laboratory supplements with a yield of 0.44 g/g. Importantly, the acid production parameters in the waste medium reached almost the same level (12.71 g/L, 0.42 g/g) as the medium containing pure supplements. Acetic acid synthesis was more efficient in the waste medium; it was also characterized by a higher level of accumulated trehalose (59.8 mg/g d.s.). Thus, the obtained results show that P. freudenreichii bacteria exhibited relatively high metabolic activity in an environment with apple pomace used as a carbon source and potato wastewater used as a nitrogen source. This method of propioniate production could be cheaper and more sustainable than the chemical manner.

Characteristics of the Proteolytic Enzymes Produced by Lactic Acid Bacteria.

Over the past several decades, we have observed a very rapid development in the biotechnological use of lactic acid bacteria (LAB) in various branches of the food industry. All such areas of activity of these bacteria are very important and promise enormous economic and industrial successes. LAB are a numerous group of microorganisms that have the ability to ferment sugars into lactic acid and to produce proteolytic enzymes. LAB proteolytic enzymes play an important role in supplying cells with the nitrogen compounds necessary for their growth. Their nutritional requirements in this regard are very high. Lactic acid bacteria require many free amino acids to grow. The available amount of such compounds in the natural environment is usually small, hence the main function of these enzymes is the hydrolysis of proteins to components absorbed by bacterial cells. Enzymes are synthesized inside bacterial cells and are mostly secreted outside the cell. This type of proteinase remains linked to the cell wall structure by covalent bonds. Thanks to advances in enzymology, it is possible to obtain and design new enzymes and their preparations that can be widely used in various biotechnological processes. This article characterizes the proteolytic activity, describes LAB nitrogen metabolism and details the characteristics of the peptide transport system. Potential applications of proteolytic enzymes in many industries are also presented, including the food industry.

Propionic acid production from apple pomace in bioreactor using Propionibacterium freudenreichii: an economic analysis of the process.

Propionic acid and its salts are widely used as food and feed preservative. Currently, these compounds are chemically produced, which is more profitable compared to biotechnological production using bacteria of the Propionibacterium genus. Appropriate steps can enable reducing the production costs; for example, cheap industrial byproducts can be used as culture media. One such cost-effective raw material is apple pomace, a low-value byproduct from the food industry. It contains sugars such as glucose and fructose which can serve as potential carbon sources for microorganisms. This paper discusses the possibility of using apple pomace in the production of propionic acid and presents an economic analysis of the production process. The tested strain produced 8.01 g/L of propionic acid (yield 0.40 g/g) and 2.29 g/L of acetic acid (yield 0.11 g/g) from apple pomace extract. The economic analysis showed that the production of 1 kg of propionic acid (considering only waste) from 1000 kg of apple pomace would cost approximately 1.25 USD. The manufacturing cost (consumables, including feedstock, labor, and utilities) would be approximately 2.35 USD/kg, and the total cost including taxes would be approximately 3.05 USD/kg. From the economic point of view, it is necessary to improve the production of propionic acid from apple pomace, to increase the yield of fermentation and thus decrease the total production costs. This can be achieved, for example, using industrial byproducts as nitrogen and vitamin sources, instead of high-cost substrates such as yeast extract or peptone. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-020-02582-x.

Metabolic Response of the Yeast Candida utilis During Enrichment in Selenium.

Selenium (Se) was found to inhibit the growth of the yeast Candida utilis ATCC 9950. Cells cultured in 30 mg selenite/L supplemented medium could bind 1368 µg Se/g of dry weight in their structures. Increased accumulation of trehalose and glycogen was observed, which indicated cell response to stress conditions. The activity of antioxidative enzymes (glutathione peroxidase, glutathione reductase, thioredoxin reductase, and glutathione S-transferase) was significantly higher than that of the control without Se addition. Most Se was bound to water-insoluble protein fraction; in addition, the yeast produced 20-30 nm Se nanoparticles (SeNPs). Part of Se was metabolized to selenomethionine (10%) and selenocysteine (20%). The HPLC-ESI-Orbitrap MS analysis showed the presence of five Se compounds combined with glutathione in the yeast. The obtained results form the basis for further research on the mechanisms of Se metabolism in yeast cells.

Biological Activity of Some Aromatic Plants and Their Metabolites, with an Emphasis on Health-Promoting Properties.

The biological activities of four aromatic plants, namely frankincense, myrrh, ginger, and turmeric, were reviewed in the current study. The volatile fraction (essential oil) as well as the nonvolatile fraction of these four plants showed different promising biological activities that are displayed in detail. These activities can include protection from and/or alleviation of some ailment, which is supported with different proposed mechanisms of action. This review aimed to finally help researchers to get a handle on the importance of considering these selected aromatic plants, which have not been thoroughly reviewed before, as a potential adjuvant to classical synthetic drugs to enhance their efficiency. Moreover, the results elicited in this review encourage the consumption of these medicinal plants as an integrated part of the diet to boost the body's overall health based on scientific evidence.

Sequencing and Analysis of the Genome of Propionibacterium Freudenreichii T82 Strain: Importance for Industry.

The genome of Propionibacterium freudenreichii ssp. freudenreichii T82, which has a chromosome containing 2,585,340 nucleotides with 67.3% GC content (guanine-cytosine content), is described in this paper. The total number of genes is 2308, of which 2260 are protein-coding genes and 48 are RNA genes. According to the genome analysis and the obtained results, the T82 strain can produce various compounds such as propionic acid, trehalose, glycogen, and B group vitamins (e.g., B6, B9, and B12). From protein-coding sequences (CDSs), genes related to stress adaptation, biosynthesis, metabolism, transport, secretion, and defense machinery were detected. In the genome of the T82 strain, sequences corresponding to the CRISPR loci (Clustered Regularly Interspaced Short Palindromic Repeats), antibiotic resistance, and restriction-modification system were found.

The aspects of microbial biomass use in the utilization of selected waste from the agro-food industry.

Cellular biomass of microorganisms can be effectively used in the treatment of waste from various branches of the agro-food industry. Urbanization processes and economic development, which have been intensifying in recent decades, lead to the degradation of the natural environment. In the first half of the 20th century, problems related to waste management were not as serious and challenging as they are today. The present situation forces the use of modern technologies and the creation of innovative solutions for environmental protection. Waste of industrial origin are difficult to recycle and require a high financial outlay, while the organic waste of animal and plant origins, such as potato wastewater, whey, lignin, and cellulose, is dominant. In this article, we describe the possibilities of using microorganisms for the utilization of various waste products. A solution to reduce the costs of waste disposal is the use of yeast biomass. Management of waste products using yeast biomass has made it possible to generate new metabolites, such as ß-glucans, vitamins, carotenoids, and enzymes, which have a wide range of industrial applications. Exploration and discovery of new areas of applications of yeast, fungal, and bacteria cells can lead to an increase in their effective use in many fields of biotechnology.

Optimization of propionic acid production in apple pomace extract with Propionibacterium freudenreichii.

Sequential optimization of propionate production using apple pomace was studied. All experiments were performed in a static flask in anaerobic conditions. Effect of apple pomace as nitrogen source against conventional N sources (yeast extract, peptone) was studied. The double increase was observed in propionic acid production while using yeast extract and peptone (0.29 ± 0.01 g/g), as against the use of only apple pomace extract (APE) (0.14 ± 0.01 g/g). Intensification of propionic acid fermentation was also achieved by increasing the pH control frequency of the culture medium from 24-(0.29 ± 0.01 g/g) to 12-hour intervals (30 °C) (0.30 ± 0.02 g/g) and by increasing the temperature of the culture from 30 to 37 °C (12-hour intervals of pH control) (0.32 ± 0.01 g/g). An important factor in improving the parameters of fermentation was the addition of biotin to the medium. The 0.2 mg/L dose of biotin allowed to attain 7.66 g/L propionate with a yield of 0.38 ± 0.03 g/g (12-hour intervals of pH control, 37 °C).

Evaluation of antioxidant and antimicrobial activity of oregano (Origanum vulgare L.) preparations during storage of low-pressure mechanically separated meat (BAADER meat) from chickens.

The goal of this study was to determine the effect of Origanum vulgare L. (oregano) preparations on the storage stability of vacuum-packed low-pressure mechanically separated meat (BAADER meat) from chickens stored at - 18 °C for 9 months. Oregano was added into the meat as a dried spice, extracts in water and ethanol (40 and 70% (v/v)), and an essential oil. The control samples did not contain oregano. The samples were examined immediately after arrival into the laboratory and after 1, 2, 3, 4, 6, and 9 months of storage. Oregano essential oil was most effective in slowing down lipid oxidation and inhibiting the growth of bacteria in BAADER meat. The number of coliform bacteria in the BAADER meat samples with the 0.1% essential oil was significantly lower than that in the control samples. The storage time was seen to have a more significant effect on the quality of BAADER meat.

Equilibrium modeling of selenium binding from aqueous solutions by Candida utilis ATCC 9950 yeasts.

The study investigated the effectiveness of selenium binding from its salt solution by Candida utilis ATCC 9950 yeast biomass cultured on a medium prepared from the agro-food industry wastes, containing an available source of carbon and nitrogen. Selenium binding by C. utilis yeast strain after 48 h of culturing at 28 °C from aqueous solutions with the addition of 30 mg Se/L reached a value of 2.28 mg Se/g of yeast biomass. The kinetics of selenium binding by the yeasts showed a better fit for the pseudo-second-order kinetic model compared to the pseudo-first-order one. Accumulation stability data were analyzed using the Freundlich and Langmuir isotherm models. The presence of competing anions such as SO42- , and HPO42- at a concentration of 0.5 mM resulted in about 35% reduction of selenium binding by the examined C. utilis strain. FTIR analysis showed that sulfur compounds were involved in selenium biosorption by the yeast. Compounds containing ammonium groups appeared to be very important for selenium binding. The results of the study demonstrated that the yeast can be used to effectively bind selenium from aqueous solution. At the same time, it gives the opportunity to obtain a biomass rich in this deficient element, which can also be used in dietary supplement production.

Propionibacterium spp.-source of propionic acid, vitamin B12, and other metabolites important for the industry.

Bacteria from the Propionibacterium genus consists of two principal groups: cutaneous and classical. Cutaneous Propionibacterium are considered primary pathogens to humans, whereas classical Propionibacterium are widely used in the food and pharmaceutical industries. Bacteria from the Propionibacterium genus are capable of synthesizing numerous valuable compounds with a wide industrial usage. Biomass of the bacteria from the Propionibacterium genus constitutes sources of vitamins from the B group, including B12, trehalose, and numerous bacteriocins. These bacteria are also capable of synthesizing organic acids such as propionic acid and acetic acid. Because of GRAS status and their health-promoting characteristics, bacteria from the Propionibacterium genus and their metabolites (propionic acid, vitamin B12, and trehalose) are commonly used in the cosmetic, pharmaceutical, food, and other industries. They are also used as additives in fodders for livestock. In this review, we present the major species of Propionibacterium and their properties and provide an overview of their functions and applications. This review also presents current literature concerned with the possibilities of using Propionibacterium spp. to obtain valuable metabolites. It also presents the biosynthetic pathways as well as the impact of the genetic and environmental factors on the efficiency of their production.

Research on the ability of propionic acid and vitamin B12 biosynthesis by Propionibacterium freudenreichii strain T82.

The purpose of this study was to determine the potential for biosynthesis of propionic acid and vitamin B12 by Propionibacterium freudenreichii T82 in a medium containing various sources of carbon (glucose, fructose, and saccharose). These sugars are present in apple pomaces, which are the waste from the production of apple juice. Using statistical analysis design of experiments (DoE), the results allowed us to determine which sugars (carbon sources) exert the most beneficial influence on the biosynthesis of propionic acid and cobalamin. The highest production of propionic acid by the tested bacterial strain was obtained in a medium in which glucose accounted for at least 50% of the available carbon sources. Depending on the culture medium, the concentration of this metabolite ranged from 23 to 40 g/L. P. freudenreichii T82 produced the smallest amount of acid in medium in which the dominant nutrient source was saccharose. The results obtained indicated an inverse relationship between the amount of acid produced by the bacteria and vitamin B12 biosynthesis. Because of the high efficiency of propionic acid biosynthesis by P. freudenreichii T82, the prospect of using this strain to obtain propionate with the simultaneous disposal of waste materials (such as apple pomaces) which contain glucose and/or fructose is very promising.

Possibility of using apple pomaces in the process of propionic-acetic fermentation

Background: In 2014, apple production in EU countries amounted to 11.8 million tonnes. A constant increase in the production of these fruits will lead to the accumulation of thousands of tonnes of apple pomace (production waste). The amount of industrial apples is the highest - their proportiononthe market is estimated at 50-60%, of which over 95% is processed into juice. The proportion of pomace in the traditional pressing method accounts for 20% offruits used. Results: Analysis of the growth dynamics of wild strain Propionibacterium freudenreichii T82 in micro-cultures using different carbon sources showed that the highest bacterial growth occurs in an environment with fructose and the most intense biosynthesis of metabolites was found in medium containing only saccharose. It has been found that P. freudenreichii T82 used apple pomaces as a source of carbon. Propionic acid biosynthesis reached its maximum value in the 120th hour of cultivation (1.771 g/L). At this time, the content of the acetic acid produced reached the level of 7.049 g/L. Conclusions: Utilization of by-products is a significant challenge for manufacturing sites and the natural environment. The solution to this problem may involve the use of pomace as a medium component for microorganism cultivation, which is a source of industrially useful metabolites. This study examined the possibility of using apple pomace as a carbon source in the process of propionic-acetic fermentation via wild strain Propionibacterium freudenreichii T82 bacteria.