Biotechnological Production Research Articles

Induction of point and structural mutations in engineered yeast Saccharomyces cerevisiae improve carotenoid production.

β-Carotene is an attractive compound and that its biotechnological production can be achieved by using engineered Saccharomyces cerevisiae. In a previous study, we developed a technique for the efficient establishment of diverse mutants through the introduction of point and structural mutations into the yeast genome. In this study, we aimed to improve β-carotene production by applying this mutagenesis technique to S. cerevisiae strain that had been genetically engineered for β-carotene production. Point and structural mutations were introduced into β-carotene-producing engineered yeast. The resulting mutants showed higher β-carotene production capacity than the parental strain. The top-performing mutant, HP100_74, produced 37.6mg/L of β-carotene, a value 1.9 times higher than that of the parental strain (20.1mg/L). Gene expression analysis confirmed an increased expression of multiple genes in the glycolysis, mevalonate, and β-carotene synthesis pathways. In contrast, expression of ERG9, which functions in the ergosterol pathway competing with β-carotene production, was decreased in the mutant strain. The introduction of point and structural mutations represents a simple yet effective method for achieving mutagenesis in yeasts. This technique is expected to be widely applied in the future to produce chemicals via metabolic engineering of S. cerevisiae.

PHBV cycle of life using waste as a starting point: from production to recyclability

Global concern about plastic pollution is forcing new policies and modifications of human consumption as well as promoting new research lines aiming at the replacement of non-degradable plastics with other polymers more environmentally friendly. Addressing food waste and promoting circular economy strategies, among other approaches, are crucial in reducing environmental impacts and fostering sustainability in several sectors like the agri-food industry. The European Union’s Circular Economy Action Plan is a significant initiative in this direction. Biotechnological processes, especially the valorisation of agri-food waste to produce highly marketed biomolecules like poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) using microorganisms as cellular factories, offer promising avenues for achieving these goals. PHBV is a biodegradable polymer firstly characterised as an isolated biopolymer from bacterial biomass. This biopolymer shows interesting physicochemical properties making possible immense potential in various applications due to its biocompatibility and sustainability, thus revealing it as a good candidate to replace plastics produced by chemical synthesis from petroleum (which are highly recalcitrant and consequently pollutants). This review critically analyses the PHBV synthesis and end-of-life scenarios from their synthesis using chemical and biological pathways, through the forms of biotechnological operation and production, to the forms described until the moment of recycling.

Isolation and characterization of a phage collection against Pseudomonas putida.

The environmental bacterium, Pseudomonas putida, possesses a broad spectrum of metabolic pathways. This makes it highly promising for use in biotechnological production as a cell factory, as well as in bioremediation strategies to degrade various aromatic pollutants. For P. putida to flourish in its environment, it must withstand the continuous threats posed by bacteriophages. Interestingly, until now, only a handful of phages have been isolated for the commonly used laboratory strain, P. putida KT2440, and no phage defence mechanisms have been characterized. In this study, we present a new Collection of Environmental P. putida Phages from Estonia, or CEPEST. This collection comprises 67 double-stranded DNA phages, which belong to 22 phage species and 9 phage genera. Our findings reveal that most phages in the CEPEST collection are more infectious at lower temperatures, have a narrow host range, and require an intact lipopolysaccharide for P. putida infection. Furthermore, we show that cryptic prophages present in the P. putida chromosome provide strong protection against the infection of many phages. However, the chromosomal toxin-antitoxin systems do not play a role in the phage defence of P. putida. This research provides valuable insights into the interactions between P. putida and bacteriophages, which could have significant implications for biotechnological and environmental applications.

Distribution and Level of Bioactive Monoacylglycerols in 12 Marine Microalgal Species.

Microalgae are currently considered an attractive source of highly valuable metabolites potentially exploitable as anticancer agents, nutraceuticals and cosmeceuticals and for bioenergy purposes. Their ease of culturing and their high growth rates further promote their use as raw material for the production of specialty products. In the present paper, we focused our attention on specific glycerol-based lipid compounds, monoacylglycerols (MAGs), which displayed in our previous studies a selective cytotoxic activity against the haematological U-937 and the colon HCT-116 cancer cell lines. Here, we performed a quali/quantitative analysis of MAGs and total fatty acids (FAs) along with a profiling of the main lipid classes in a panel of 12 microalgal species, including diatoms and dinoflagellates. Our results highlight an inter- and intraspecific variability of MAG profile in the selected strains. Among them, Skeletonema marinoi (strain FE7) has emerged as the most promising source for possible biotechnological production of MAGs.

The Progress of the Biotechnological Production of Class IIa Bacteriocins in Various Cell Factories and Its Future Challenges.

Class IIa bacteriocins produced in lactic acid bacteria are short cationic peptides with antimicrobial activity. In the search for new biopreservation agents, class IIa bacteriocins are considered to be the best potential candidates, not only due to their large abundance but also because of their high biological activity and excellent thermal stability. However, regulated by the biosynthetic regulatory system, the natural class IIa bacteriocin yield is low, and the extraction process is complicated. The biotechnological production of class IIa bacteriocins in various cell factories has been attempted to improve this situation. In this review, we focus on the application of biotechnological routes for class IIa bacteriocin production. The drawbacks and improvements in the production of class IIa bacteriocins in various cell factories are discussed. Furthermore, we present the main challenge of class IIa bacteriocins, focusing on increasing their production by constructing suitable cell factories. Recombinant bacteriocins have made considerable progress from inclusion body formation, dissolved form and low antibacterial activity to yield recovery. The development of prospective cell factories for the biotechnological production of bacteriocins is still required, which may facilitate the application of bacteriocins in the food industry.

The master key: structural science in unlocking functional materials advancements

From the historical roots of metalworking to the forefront of modern nanotechnology, functional materials have played a pivotal role in transforming societies, and their influence is poised to persist into the future. Encompassing a wide array of solid-state materials, spanning semiconductors to polymers, molecular crystals to nanoparticles, functional materials find application in critical sectors such as electronics, computers, information, communication, biotechnology, aerospace, defense, environment, energy, medicine and consumer products. This feature article delves into diverse instances of functional materials, exploring their structures, their properties and the underlying mechanisms that contribute to their outstanding performance across fields like batteries, photovoltaics, magnetics and heterogeneous catalysts. The field of structural sciences serves as the cornerstone for unraveling the intricate relationship between structure, dynamics and function. Acting as a bridge, it connects the fundamental understanding of materials to their practical applications.

A new Zymomonas mobilis platform strain for the efficient production of chemicals

BackgroundZymomonas mobilis is well known for its outstanding ability to produce ethanol with both high specific productivity and with high yield close to the theoretical maximum. The key enzyme in the ethanol production pathway is the pyruvate decarboxylase (PDC) which is converting pyruvate to acetaldehyde. Since it is widely considered that its gene pdc is essential, metabolic engineering strategies aiming to produce other compounds derived from pyruvate need to find ways to reduce PDC activity.ResultsHere, we present a new platform strain (sGB027) of Z. mobilis in which the native promoter of pdc was replaced with the IPTG-inducible PT7A1, allowing for a controllable expression of pdc. Expression of lactate dehydrogenase from E. coli in sGB027 allowed the production of D-lactate with, to the best of our knowledge, the highest reported specific productivity of any microbial lactate producer as well as with the highest reported lactate yield for Z. mobilis so far. Additionally, by expressing the L-alanine dehydrogenase of Geobacillus stearothermophilus in sGB027 we produced L-alanine, further demonstrating the potential of sGB027 as a base for the production of compounds other than ethanol.ConclusionWe demonstrated that our new platform strain can be an excellent starting point for the efficient production of various compounds derived from pyruvate with Z. mobilis and can thus enhance the establishment of this organism as a workhorse for biotechnological production processes.

ALTERNATIVE NON-TRADITIONAL SOURCES OF FEED PROTEIN AND AMINO ACIDS

The article presents a literature review on the risks of the global problem of fodder protein and protein amino acids in the feeding of agricultural animals and poultry. Alternative ways of solving it are due to the production and use of non-traditional feed raw materials in feed diets. A number of scientists consider atypical agricultural biotechnological products obtained from insects to be one of the alternative sources of fodder protein and amino acids and see prospects in the development of their production. The most common subjects for study in this regard are crickets, locusts, silkworm larvae, flies, worms, etc. Their protein value, amino acid, fatty acid and mineral composition, the content of vitamins, and other biologically active substances are studied. The development of this direction is considered one of the ways to solve a number of urgent environmental and fodder problems on a biological basis. Рurpose of the research was to study the protein value and complete amino acid composition of flour from insects, namely flour from the larvae of the BSF fly, crickets and snails, and the methods of its determining: crude protein by the Kjeldahl method, digestible protein with the help of 0,2% pepsin, and the digestibility coefficient, and the principles of conducting researches, as well as the characteristics of methods for determining the content of replaceable and essential amino acids, including sulfur-containing, using the Kapel-105M capillary electrophoresis system. As a result of the research, it was found that cricket flour is the least valuable in terms of protein value (digestible protein and digestibility coefficient), and it is also poor in the content of nonessential and essential amino acids, compared to other studied types of insect flour. The best indicators of protein value were noted in the flour from BSF fly larvae in terms of the content of crude and digestible proteins, as well as the digestibility coefficient, which is 75.6%, which indicates its good digestibility by the bodies of animals and birds. The highest content of amino acids is contained in the flour from BSF fly larvae. Taking into account the above results of crude protein content, digestibility, and digestibility in this sample, amino acid digestibility will also be higher compared to other flour samples. In addition, flour from BSF fly larvae is rich in sulfur-containing amino acids. The content of tryptophan and histidine, compared to other types of raw materials, is the highest. The studied samples of insect flour contain a complete set of essential and nonessential amino acids, but BSF fly larvae flour and snail flour are more valuable for essential amino acids.

Paclitaxel - a Product of Fungal Secondary Metabolism or an Artefact?

Taxol (common name: paclitaxel) is an extremely important component of drugs for the treatment of various cancers. Thirty years after the discovery of its effectiveness, a metabolic precursor of Taxol (10-deacetylbaccatin III) is still primarily extracted from needles of European yew trees. In order to meet the considerable demand, hopes were pinned on the possibilities of biotechnological production from the very beginning. In 1993, as if by chance, Taxol was supposedly discovered in fungi that grow endobiotically in yew trees. This finding aroused hopes of biotechnological use to produce fungal Taxol in large quantities in fermenters. It never came to that. Instead, a confusing flood of publications emerged that claimed to have detected Taxol in more and more eukaryotic and even prokaryotic species. However, researchers never reproduced these rather puzzling results, and they could certainly not be applied on an industrial scale. This paper will show that some of the misguided approaches were apparently based on a seemingly careless handling of sparse evidence and on at least questionable publications. Apparently, the desired gold rush of commercial exploitation was seductive. Scientific skepticism as an indispensable core of good scientific practice was often neglected, and the peer review process has not exerted its corrective effect. Self-critical reflection and more healthy skepticism could help to reduce the risk of such aberrations in drug development. This article uses this case study as a striking example to show what can be learned from the Taxol case in terms of research ethics and the avoidance of questionable research practices.

Antioxidant and Anticancer Potential of Extracellular Polysaccharide from Porphyridium aerugineum (Rhodophyta)

Porphyridium aerugineum is a unicellular freshwater red microalga that synthesizes and secretes into the culture medium an extracellular polysaccharide (EPS). In this study, algal growth and polysaccharide production, as well as the antioxidant capacity and antitumor effect of Porphyridium aerugineum EPS (PaEPS), were investigated. Cultivation of the microalgae was carried out in a photobioreactor under controlled conditions. Algal growth and the amount of EPS were monitored daily. The accumulated polysaccharide was extracted and lyophilized. At the end of cultivation, the concentration of microalgal biomass and PaEPS reached 3.3 and 1.2 g L−1, respectively. To examine the antioxidant capacity of PaEPS, FRAP and ABTS assays were performed. The cytotoxic activity of PaEPS was evaluated on the tumor cell lines MCF-7 (breast cancer) and HeLa (cervical adenocarcinoma) and on BJ (a non-tumor human skin fibroblast cell line), using MTT assay. The results obtained indicated that P. aerugineum polysaccharide exhibited a high ABTS radical-scavenging activity reaching up to 55%. The cytotoxic effect was best expressed in MCF-7 cells treated for 72 h with 1000 µg/mL PaEPS, where tumor cell proliferation was inhibited by more than 70%. Importantly, the PaEPS treatments did not significantly affect the viability of BJ cells. These findings promote the biotechnological production of P. aerugineum extracellular polysaccharide and reveal its potential as an anticancer and antioxidant agent for future applications.

Association between organic nitrogen substrates and the optical purity of d-lactic acid during the fermentation by Sporolactobacillus terrae SBT-1

The development of biotechnological lactic acid production has attracted attention to the potential production of an optically pure isomer of lactic acid, although the relationship between fermentation and the biosynthesis of highly optically pure d-lactic acid remains poorly understood. Sporolactobacillus terrae SBT-1 is an excellent d-lactic acid producer that depends on cultivation conditions. Herein, three enzymes responsible for synthesizing optically pure d-lactic acid, including d-lactate dehydrogenase (D-LDH; encoded by ldhDs), l-lactate dehydrogenase (L-LDH; encoded by ldhLs), and lactate racemase (Lar; encoded by larA), were quantified under different organic nitrogen sources and concentration to study the relationship between fermentation conditions and synthesis pathway of optically pure lactic acid. Different organic nitrogen sources and concentrations significantly affected the quantity and quality of d-lactic acid produced by strain SBT-1 as well as the synthetic optically pure lactic acid pathway. Yeast extract is a preferred organic nitrogen source for achieving high catalytic efficiency of d-lactate dehydrogenase and increasing the transcription level of ldhA2, indicating that this enzyme plays a major role in d-lactic acid formation in S. terrae SBT-1. Furthermore, lactate racemization activity could be regulated by the presence of d-lactic acid. The results of this study suggest that specific nutrient requirements are necessary to achieve a stable and highly productive fermentation process for the d-lactic acid of an individual strain.

Mastery concepts and misconceptions in biotechnology courses with a three tier diagnostic test

This research aims to determine the mastery of concepts and misconceptions of students in the Biology Education Study Program wich is in Pekanbaru. This type of research is survey research with a porpusive sampling type. The population of the study was all students who had taken biotechnology courses, while the research sample was 6th semester students who had just completed biotechnology courses in the previous semester. The total research sample was 32 students. The instrument used to measure mastery of concepts and misconceptions is a diagnostic test with 20 multiple choice questions accompanied by reasons and level of belief. The questions are given using a Google form and students work on the questions during class and are given 90 minutes.The research data were analyzed descriptively. Based on the results showed that for indicator 1 30.01% of students understood the basic concepts of biotechnology well and 55.63 had misconceptions. In indicator 2, only 5% of students understood the concept of biotechnology process while 64.38% had misconceptions and 29.38 did not understand the concept. Indicator 3, as many as 10.63% of students understand the concept of conventional and modern biotechnology characteristics and 59.38% of students have misconceptions and 25.63% do not understand the concept while indicator 4 as many as 30.63% of students understand the examples of biotechnology products while still 43.13% have misconceptions and 22.50% still do not understand the concept. Students' mastery of concepts in biotechnology courses is in a very low category and while more than half of the students still experience misconceptions and do not understand the concepts.

Taxonomic report of filamentous fungi presents in sewage waste sludge with biotechnologi-cal potential

There is few information regarding the identification of filamentous fungi present in sewage sludge, the importance of these microorganisms lies in the potential use they have in the food, pharmaceutical and agri-environmental industries as they are beneficial in the remediation of water, sludge and soil contaminated by Heavy metals and hydrocarbons also contribute to the adsorption of nutrients by crops. The use of residual sludge once decontaminated by these microorganisms can be used in agriculture as soil structure improvers, organic fertilizers and as an amendment to prevent soil erosion. The objective of this research was to characterize and identify fast and medium-growing filamentous fungi from sewage sludge. The residual sludge was collected directly from the thickness of the municipal water treatment plant of Cd. Victoria, Tamaulipas, Mexico. Subsequently, the fungi were isolated on agar plates and identified using morphological characters of the colonies and by PCR and diana sequences. The species identified were: Trichoderma longibrachiatum, Aspergillus terreus, Tylopilus porphyrosporus and Aspergillus fumigatus. Being the first report for Mexico of fungal species that grow in residual sludge from water treatment plants. The molecular characterization of the species of filamentous fungi found in sewage sludge will allow progress in research focused on knowing the advantages and biological and chemical mechanisms of these microorganisms for the development of biotechnological processes and products, with high agricultural potential and environmental.

Ultrasound-guided accesses for regenerative injection therapy of hip and knee

Interventional diagnostic and therapeutic technologies have gained considerable popularity in many surgical specialties – cardiology, vascular surgery, neurosurgery, oncology, and in orthopedics and traumatology. But unlike other specialties, ultrasound became the driving force behind the development and implementation of interventional technologies in the treatment of diseases of the musculoskeletal system. In recent years, ultrasound-guided injections have gained significant popularity, as they have given an advantage in accuracy compared to injections without such navigation. The purpose of our study was the development of ultrasound navigation accesses to the hip and knee joints for the interventional application of regenerative technologies in their pathology. The material for the study was the results of ultrasound examination of the hip and knee joints of 486 patients with diseases and injuries of the hip and knee, who were treated in the scientific and practical department of tissue and cell therapy of the State Institute of Traumatology and Orthopedics of the National Academy of Medical Sciences of Ukraine in the period from 2016 to 2023. The results. The following accesses to the knee joint under ultrasound navigation have been developed: suprapatellar longitudinal access for injections into the quadriceps tendon and patellofemoral joint, suprapatellar longitudinal access with knee bending for injections into the upper turn of the knee joint, infrapatellar longitudinal access for injections into the patellar ligament and deep infrapatellar bursa, infrapatellar transverse access for injections into Hoffa’s fat pad, lateral longitudinal access with knee bending for injections into the lateral meniscus, medial longitudinal access with bending in the knee for injections in the medial meniscus, medial transverse access for injections in the medial part of the joint space, lateral transverse access with bending in the knee joint for injections in the lateral part of the joint space, lateral longitudinal access for performing injections in the collateral fibular ligament, medial longitudinal access for performing injections in the collateral tibial ligament, infrapatellar diagonal access for performing injections in the “crow’s foot” area of the knee joint, infrapatellar medial longitudinal access for performing other injection into the medial meniscus and joint capsule, lateral longitudinal access for injections into the tendons of the biceps femoris and hamstrings. The following accesses have been developed for the hip joint: anterior longitudinal access for performing injections in the subcapsular-cervical space, anterior diagonal access for performing injections in the acetabular labrum, capsule-ligament apparatus of the hip and tendon of the rectus femoris muscle, lateral longitudinal access for performing injections in the paratrochanteric region. Conclusions. 13 ultrasonic navigation accesses have been developed for the administration of biotechnological products into the knee joint and 3 – into the hip join, which allow not only to detect damaged intra-articular and para-articular structures of the knee and hip joints, but also to deliver an orthobiologics products directly to them for maximum regenerative effect.

Systems metabolic engineering of Corynebacterium glutamicum to assimilate formic acid for biomass accumulation and succinic acid production

Formate as an ideal mediator between the physicochemical and biological realms can be obtained from electrochemical reduction of CO2 and used to produce bio-chemicals. Yet, limitations arise when employing natural formate-utilizing microorganisms due to restricted product range and low biomass yield. This study presents a breakthrough: engineered Corynebacterium glutamicum strains (L2-L4) through modular engineering. L2 incorporates the formate-tetrahydrofolate cycle and reverse glycine cleavage pathway, L3 enhances NAD(P)H regeneration, and L4 reinforces metabolic flux. Metabolic modeling elucidates C1 assimilation, guiding strain optimization for co-fermentation of formate and glucose. Strain L4 achieves an OD600 of 0.5 and produces 0.6 g/L succinic acid. 13C-labeled formate confirms C1 assimilation, and further laboratory evolution yields 1.3 g/L succinic acid. This study showcases a successful model for biologically assimilating formate in C. glutamicum that could be applied in C1-based biotechnological production, ultimately forming a formate-based bioeconomy.

Toxicity and Teratogenic Potential of Piplartine from Piper tuberculatum Jacq. during Embryonic Development in Mice (Mus musculus)

Piplartine, also known as piperlongumine, is a natural and biologically active amide alkaloid found in various Piper species within the Piperaceae family. It possesses numerous beneficial properties that can be leveraged in the development of nanotechnological and pharmaceutical products. However, information on the effects of piplartine on mammalian embryonic development is scarce. This study aims to assess the general toxicity and teratogenic potential of piplartine during the embryonic development of mice. Pregnant mice received daily treatments of 25, 50, or 100 mg/kg of piplartine via gavage from the sixth day of gestation (implantation) to the eighteenth. On the eighteenth day, the mice were euthanized, and whole organs, blood samples (for hematological and biochemical analyses), and bone marrow cells (for DNA fragmentation and cell cycle assays) were collected. The uterus was examined for implantation sites and embryo resorptions. Additionally, fetuses were collected to assess for fetal anomalies. Piplartine did not result in maternal or embryo-fetal toxicity, induce fetal anomalies, cause hematological and biochemical alterations, or lead to DNA fragmentation. The oral administration of piplartine is safe and does not exhibit toxicity or teratogenic effects in mice. This finding opens avenues for the development of piplartine-based biotechnological products for therapeutic interventions in disease treatment.

Therapeutic Effects of Essential Oils and Their Bioactive Compounds on Prostate Cancer Treatment.

Since prostate cancer (PCa) relies on limited therapies, more effective alternatives are required. Essential oils (EOs) and their bioactive compounds are natural products that have many properties including anticancer activity. This review covers studies published between 2000 and 2023 and discusses the anti-prostate cancer mechanisms of the EOs from several plant species and their main bioactive compounds. It also provides a critical perspective regarding the challenges to be overcome until they reach the market. EOs from chamomile, cinnamon, Citrus species, turmeric, Cymbopogon species, ginger, lavender, Mentha species, rosemary, Salvia species, thyme and other species have been tested in different PCa cell lines and have shown excellent results, including the inhibition of cell growth and migration, the induction of apoptosis, modulation in the expression of apoptotic and anti-apoptotic genes and the suppression of angiogenesis. The most challenging aspects of EOs, which limit their clinical uses, are their highly lipophilic nature, physicochemical instability, photosensitivity, high volatility and composition variability. The processing of EO-based products in the pharmaceutical field may be an interesting alternative to circumvent EOs' limitations, resulting in several benefits in their further clinical use. Identifying their bioactive compounds, therapeutic effects and chemical structures could open new perspectives for innovative developments in the field. Moreover, this could be helpful in obtaining versatile chemical synthesis routes and/or biotechnological drug production strategies, providing an accurate, safe and sustainable source of these bioactive compounds, while looking at their use as gold-standard therapy in the close future.

Advances in biotin biosynthesis and biotechnological production in microorganisms.

Biotin, also known as vitamin H or B7, acts as a crucial cofactor in the central metabolism processes of fatty acids, amino acids, and carbohydrates. Biotin has important applications in food additives, biomedicine, and other fields. While the ability to synthesize biotin de novo is confined to microorganisms and plants, humans and animals require substantial daily intake, primarily through dietary sources and intestinal microflora. Currently, chemical synthesis stands as the primary method for commercial biotin production, although microbial biotin production offers an environmentally sustainable alternative with promising prospects. This review presents a comprehensive overview of the pathways involved in de novo biotin synthesis in various species of microbes and insights into its regulatory and transport systems. Furthermore, diverse strategies are discussed to improve the biotin production here, including mutation breeding, rational metabolic engineering design, artificial genetic modification, and process optimization. The review also presents the potential strategies for addressing current challenges for industrial-scale bioproduction of biotin in the future. This review is very helpful for exploring efficient and sustainable strategies for large-scale biotin production.

Acetic Fermentation of Cagaita Pulp: Technological and Chemical Characteristics

The Brazilian Cerrado region has a rich plant diversity, with fruits that have peculiar and unique sensory characteristics. For these reasons, using these fruits for biotechnological production is a promising alternative, mainly to protect this biome from deforestation and degradation. The production of fermented acetic acid is an option to add value to native fruits and offer the market beverages with better nutritional quality and bioactive compounds. This work aimed to characterize fruits and to develop cagaita (Eugenia dysenterica DC.) acetic fermented beverage. The fruits were subjected to physical-chemical analyses in the first part. Subsequently, different treatments for fermentation were tested using two types of enzymes (amylase and pectinase), two subspecies of Saccharomyces cerevisiae yeast (UFLA CA11 and thermoresistant LNF Angel), and the chaptalization of the must with sucrose (16 °Brix). Alcoholic fermentation was carried out in an incubator with temperature control at 34 ± 1 °C. The pH, total soluble solids, titratable acidity, alcohol content, and density of the fermented products were monitored daily. The chaptalized must with amylase addition and thermoresistant yeast had the best performance during alcoholic fermentation, demonstrating that thermoresistant yeast is an economically advantageous and efficient alternative for the cagaita juice fermentation process. Subsequently, acetic fermentation was carried out using the slow method. Heat-resistant yeast without added enzymes was used to produce cagaita acetic fermented beverages within the parameters of the Brazilian legislation. Furthermore, phenolic compounds and antioxidant activity in the final product were observed. The work demonstrated the possibility of using cagaita fruits in biotechnological processes to produce new food products.

Potential Pharmaceutical Biotechnology Products in the Form of Formulations and Preparations of Telang Flower Kombucha Dish Soap (Clitoria ternatea L) as an Antibacterial from Beef (Bos taurus) Isolate

This research aims to see the potential of each formulation and preparation of dishwashing soap with the active ingredient telang flower kombucha in inhibiting the growth of pathogenic bacteria isolated from beef. This research is a laboratory experiment, namely by making a dishwashing soap base without the active substance of telang flower kombucha. Make a dishwashing soap base that adds the active substance of butterfly pea flower kombucha at concentrations of 20%, 30% and 40%. Biochemical identification results have shown that beef samples contain gram-positive bacteria in the form of Staphylococcus aureus and Listeria monocytogenes, as well as gram-negative bacteria in the form of Salmonella thyposa and Escherichia coli. The results of this research have also proven that pharmaceutical biotechnology products in the form of formulations and dishwashing soap preparations with the active ingredient telang flower kombucha have a positive correlation in inhibiting the growth of the four test bacteria. Based on the one-way ANOVA statistical test at each P value <0.05, it proves that telang flower kombucha dish soap at a concentration of 40% according to post hoc analysis is not significantly different from concentrations of 20% and 30% as a pharmaceutical and antibacterial biotechnology product. isolated from beef. The conclusion of this research is that the formulation and preparation of dishwashing soap at a concentration of 40% proved to be the most effective as a pharmaceutical biotechnology product in inhibiting the growth of the four test bacteria.