Microbial transglutaminase (MTG) is an enzyme widely used in the food industry because it creates cross-links between proteins, enhancing the texture and stability of food products. Its unique properties make it a valuable tool for modifying the functional characteristics of proteins, significantly impacting the quality and innovation of food products. In this study, response surface methodology was employed to optimize the fermentation conditions for microbial transglutaminase production by the strain Streptoverticillium cinnamoneum KKP 1658. The effects of nitrogen dose, cultivation time, and initial pH on the activity of the produced transglutaminase were investigated. The significance of the examined factors was determined as follows: cultivation time > nitrogen dose > pH. The interaction between nitrogen dose and cultivation time was found to be crucial, having the second most significant impact on transglutaminase activity. Optimal conditions were identified as 48 h of cultivation with a 2% nitrogen source dose and an initial medium pH of approximately 6.0. Under these conditions, transglutaminase activity ranged from 4.5 to 5.5 U/mL. The results of this study demonstrated that response surface methodology is a promising approach for optimizing microbial transglutaminase production. Future applications of transglutaminase include the development of modern food products with improved texture and nutritional value, as well as its potential use in regenerative medicine for creating biomaterials and tissue scaffolds. This topic is particularly important and timely as it addresses the growing demand for innovative and sustainable solutions in the food and biomedical industries, contributing to an improved quality of life.
The present study has undertaken the isolation of marine yeasts from mangrove sediment samples and their ability to produce alkaline protease enzymes. A total of 14 yeast isolates were recovered on yeast-malt agar (YMA) and yeast extract peptone dextrose (YEPD) agar medium. After screening for proteolytic activity on skim milk agar, marine yeast isolate, AKB-1 exhibited a hydrolysis zone of 18 mm. Optimal conditions for the enzyme production from yeast isolate AKB-1 were at 30 °C, pH 8, fructose as carbon source, potassium nitrate as nitrogen source, and 25% saline concentration. Under the optimal conditions, the protease enzyme activity of the isolate AKB-1 was observed to be 978 IU/mL. The structural and functional analysis was carried out through FTIR and HPLC analysis for the extracted protease enzyme. Furthermore, the enzyme produced was partially purified by solvent extraction using ethyl acetate and ammonium sulfate precipitation (3.4-fold) followed by dialysis (56.8-fold). The molecular weight of the purified enzyme was observed to be around 60 kDa using SDS-PAGE. The extracted protein showed good antibacterial activity against six different clinical bacterial pathogens and the highest against Bacillus cereus (16 ± 0.5 mm). The extracted protease enzyme was revealed to remove blood stains from cloth within 20 min of application similar to the commercial detergent. The marine yeast isolate was further identified as Candida orthopsilosis AKB-1 (Accession number KY348766) through 18S rRNA sequencing, and a phylogenetic tree was generated.
- MeSH
- Anti-Bacterial Agents pharmacology metabolism chemistry isolation & purification MeSH
- Bacillus cereus drug effects MeSH
- Bacterial Proteins * chemistry pharmacology metabolism isolation & purification MeSH
- Candida * enzymology isolation & purification genetics classification MeSH
- Endopeptidases * chemistry metabolism isolation & purification pharmacology MeSH
- Phylogeny MeSH
- Geologic Sediments microbiology MeSH
- Hydrogen-Ion Concentration MeSH
- Culture Media chemistry MeSH
- Microbial Sensitivity Tests MeSH
- Molecular Weight MeSH
- Enzyme Stability MeSH
- Temperature MeSH
- Publication type
- Journal Article MeSH
β-Glucans comprise a group of β-D-glucose polysaccharides (glucans) that occur naturally in the cell walls of bacteria, fungi, and cereals. Its degradation is catalyzed by β-glucanases, enzymes that catalyze the breakdown of β-glucan into cello-oligosaccharides and glucose. These enzymes are classified as endo-glucanases, exo-glucanases, and glucosidases according to their mechanism of action, being the lichenases (β-1,3;1,4-glucanases, EC 3.2.1.73) one of them. Hence, we aimed to enhance lichenase production by Thermothelomyces thermophilus through the application of response surface methodology, using tamarind (Tamarindus indica) and jatoba (Hymenaea courbaril) seeds as carbon sources. The crude extract was immobilized, with a focus on improving lichenase activity, using various ionic supports, including MANAE (monoamine-N-aminoethyl), DEAE (diethylaminoethyl)-cellulose, CM (carboxymethyl)-cellulose, and PEI (polyethyleneimine)-agarose. Regarding lichenase, the optimal conditions yielding the highest activity were determined as 1.5% tamarind seeds, cultivation at 50 °C under static conditions for 72 h. Moreover, transitioning from Erlenmeyer flasks to a bioreactor proved pivotal, resulting in a 2.21-fold increase in activity. Biochemical characterization revealed an optimum temperature of 50 °C and pH of 6.5. However, sustained stability at varying pH and temperature levels was challenging, underscoring the necessity of immobilizing lichenase on ionic supports. Notably, CM-cellulose emerged as the most effective immobilization medium, exhibiting an activity of 1.01 U/g of the derivative (enzyme plus support), marking a substantial enhancement. This study marks the first lichenase immobilization on these chemical supports in existing literature.
- MeSH
- Enzymes, Immobilized * metabolism chemistry MeSH
- Fungal Proteins * metabolism chemistry MeSH
- Glycoside Hydrolases * metabolism chemistry biosynthesis MeSH
- Hydrogen-Ion Concentration MeSH
- Fruit metabolism MeSH
- Seeds metabolism MeSH
- Sordariales MeSH
- Enzyme Stability MeSH
- Tamarindus metabolism microbiology MeSH
- Publication type
- Journal Article MeSH
Among carotenoids, ꞵ-carotene has the highest biological activity and is found as an all-trans isomer in many biological systems. Blakeslea trispora is a microorganism that is of interest to industries for the commercial production of ꞵ-carotene. This study investigated the effect of different bacteria on carotenogenesis in B. trispora. The B. trispora bisexual mold was cultured in a production medium, and different bacterial cells were added to it after 24 h. Then, the culture conditions and the culture medium were optimized in the presence of the selected bacteria using the experimental design. The percentage of carotenoids obtained from the mixed culture was determined using high-performance liquid chromatography (HPLC). Results showed that Kocuria rhizophila had the greatest effect on increasing the production of carotenoids in B. trispora. The highest content of carotenoids obtained during optimization was 770 ± 7.5 mg/L, a 6.8-fold increase compared to the control. HPLC analysis of carotenoids indicated the presence of two main peaks, ꞵ-carotene and γ-carotene, in which the primary carotenoid was ꞵ-carotene followed by γ-carotene with a lower content. Therefore, due to the importance of ꞵ-carotene in industry, the use of biostimulants is one of the appropriate strategies to increase the production of this pigment in industry.
Feruloyl esterases (FAEs) are a crucial component of the hemicellulose-degrading enzyme family that facilitates the degradation of lignocellulose while releasing hydroxycinnamic acids such as ferulic acid with high added value. Currently, the low enzyme yield of FAEs is one of the primary factors limiting its application. Therefore, in this paper, we optimized the fermentation conditions for the expression of FAE BpFaeT132C-D143C with excellent thermal stability in Escherichia coli by experimental design. Firstly, we explored the effects of 11 factors such as medium type, isopropyl-β-D-thiogalactopyranoside (IPTG) concentration, and inoculum size on BpFaeT132C-D143C activity separately by the single factor design. Then, the significance of the effects of seven factors, such as post-induction temperature, shaker rotational speed, and inoculum size on BpFaeT132C-D143C activity, was analyzed by Plackett-Burman design. We identified the main factors affecting the fermentation conditions of E. coli expressing BpFaeT132C-D143C as post-induction temperature, pre-induction period, and post-induction period. Finally, we used the steepest ascent path design and response surface method to optimize the levels of these three factors further. Under the optimal conditions, the activity of BpFaeT132C-D143C was 3.58 U/ml, which was a significant 6.6-fold increase compared to the pre-optimization (0.47 U/ml), demonstrating the effectiveness of this optimization process. Moreover, BpFaeT132C-D143C activity was 1.52 U/ml in a 3-l fermenter under the abovementioned optimal conditions. It was determined that the expression of BpFaeT132C-D143C in E. coli was predominantly intracellular in the cytoplasm. This study lays the foundation for further research on BpFaeT132C-D143C in degrading agricultural waste transformation applications.
- MeSH
- Escherichia coli * genetics metabolism enzymology MeSH
- Fermentation * MeSH
- Isopropyl Thiogalactoside metabolism MeSH
- Carboxylic Ester Hydrolases * genetics metabolism chemistry biosynthesis MeSH
- Culture Media chemistry MeSH
- Coumaric Acids metabolism MeSH
- Lignin MeSH
- Recombinant Proteins genetics metabolism biosynthesis chemistry MeSH
- Enzyme Stability MeSH
- Temperature MeSH
- Publication type
- Journal Article MeSH
Biopriming seeds with beneficial bacteria has potential to enhance seed germination. Therefore, in this investigation, five sulphur-oxidizing bacterial cultures, viz., Pantoea dispersa SOB2, Bacillus velezensis SN06, Bacillus cereus SN20, Bacillus tropicus SN16, and Bacillus megaterium SN11, were evaluated for different plant growth-promoting traits and their impact on Vigna radiata L. (mung bean) and Brassica juncea L. (mustard) seed germination. Among these, three bacterial cultures Pantoea dispersa SOB2, Bacillus velezensis SN06, and Bacillus megaterium SN11 evinced potential for mineral solubilization on solid medium where Pantoea dispersa SOB2 had the maximum solubilization indices-3.06, 5.14, and 2.48 for phosphate, zinc, and potassium respectively. The culture also displayed higher indole acetic acid (113.12 μg/mL), gibberellic acid (162.66 μg/mL), ammonia (5.23 μg/mL), and siderophore (69.53%) production than other bacterial cultures whereas Bacillus cereus SN20 showed maximum exopolysaccharide production (9.26 g/L). Bacterial culture Pantoea dispersa SOB2 significantly ameliorated the germination rate (3.73 no./day) and relative seed germination (208%) of Brassica juncea L., while Bacillus velezensis SN06 and Bacillus cereus SN20 followed with germination rate and relative seed germination of 2.86 no./day and 207%, respectively. Pantoea dispersa SOB2 displayed lowest mean germination time 2.91 days followed by Bacillus megaterium SN11 with 3.19 days. Biopriming with sulphur-oxidizing bacterial cultures, germination parameters of Vigna radiata L. were also markedly improved. Pantoea dispersa SOB2 demonstrated the highest germination rate (6.72 no./day), relative seed germination (115.56%), and minimum mean generation time (1.73 days). Bacillus velezensis SN06 inoculation had a beneficial effect on the seedling growth of Vigna radiata L., whereas Pantoea dispersa SOB2 greatly aided the seedling growth of Brassica juncea L. Results corroborated a prominent positive correlation between seed germination and plant growth-promoting traits. This is the first study on Pantoea dispersa as sulphur oxidizer, displaying plant growth promoting traits and seed germination potential. The potent sulphur-oxidizing bacterial cultures possessing plant growth promoting activities enhanced seed germination under in vitro conditions that could be further explored in field as biofertilizers to enhance the growth and yield of Brassica juncea L. and Vigna radiata L. crop.
- MeSH
- Bacillus * metabolism MeSH
- Bacteria * metabolism MeSH
- Mustard Plant * growth & development microbiology MeSH
- Germination * MeSH
- Indoleacetic Acids metabolism MeSH
- Oxidation-Reduction MeSH
- Pantoea metabolism MeSH
- Plant Growth Regulators metabolism MeSH
- Seeds * microbiology growth & development MeSH
- Sulfur * metabolism MeSH
- Vigna * growth & development microbiology MeSH
- Publication type
- Journal Article MeSH
Obrovskobunková arteritída predstavuje vaskulitídu stredných a veľkých ciev. Kazuistika opisuje prípad 56-ročnej pacientky s anamnézou chronického vertebrogénneho algického syndrómu cervikálnej chrbtice (VAS C chrbtice) s pestrým klinickým obrazom: bolesť hlavy a šije, bolesť a edém ramien, klaudikácie pri žuvaní, edém krku, febrility, nechutenstvo, dy fágia a nápadný hmotnostný úbytok. Vzhľadom na pomerne fyzicky náročnú prácu pacientky sa opätovne myslelo na vertebrogénny pôvod ťažkostí. V júni 2023 bola hospitali- zovaná na neurologickom oddelení, kde bol stav hodnotený ako recidíva VAS C chrbtice s radikulárnou iritáciou C5 bilaterálne. Avšak vzhľadom na pretrvávajúcu eleváciu zápalových parametrov bola v auguste 2023 opäť hospitalizovaná na internom oddelení. Pre suspekciu na polymyalgia rheumatica bola u nej iniciovaná liečba kortikosteroidmi v dávke 20 mg/deň, pri ktorej došlo k ústupu febrilít a poklesu zápalových parametrov. PET/CT vyšetrenie potvrdilo vaskulitídu (pri liečbe kortikosteroidmi len s miernou zápalovou aktivitou v oblastiach postihnutých ciev). Stav bol nakoniec uzavretý ako obrovskobunková arteritída s polymyalgia rheumatica. Dávka kortikosteroidov navýšená s následnou detrakciou. Vzhľadom na chýbanie postihnutia a. temporalis superficialis bioptické vyšetrenie nebolo realizované. Cieľom kazuistiky bolo poukázať na význam včasnej diagnostiky a liečby pacientov s obrovskobunkovou arteritídou, ktorá umožňuje predísť rozvoju závažných komplikácií ochorenia.
Giant cell arteritis represents vasculitis of medium and large vessels. The case report describes a 56-year-old female patient with a history of chronic vertebrogenic algic syndrome of the cervical spine (VAS C-spine) with a varied clinical picture: headache and neck pain, shoulder pain and oedema, claudication on chewing, neck oedema, febrility, inappetence, dysphagia, and conspicuous weight loss. Given the patient’s relatively physically demanding job, the vertebrogenic origin of the complaints was again thought of. In June 2023, she was admitted to the neurology department where the condition was assessed as recurrence of VAS C-spine with radicular irritation of C5 bilaterally. However, due to persistent elevation of inflammatory parameters, she was again admitted to the internal medicine department in August 2023. Due to suspicion of polymyalgia rheumatica, corticosteroid therapy at a dose of 20 mg/day was initiated, which resulted in recession of febrility and a decrease in inflammatory parameters. PET/CT scan confirmed vasculitis (with only mild inflammatory activity in the areas of affected vessels with corticosteroid treatment). The condition was finally concluded as giant cell arteritis with polymyalgia rheumatica. Corticosteroid dose increased with subsequent detraction. Biopsy examination was not performed due to the absence of involvement of the temporal artery. The aim of the case report was to highlight the importance of early diagnosis and treatment of patients with giant cell arteritis to prevent the development of serious complications of the disease.
Current antibiotics and chemotherapeutics are becoming ineffective because pathogenic bacteria and tumor cells have developed multiple drug resistance. Therefore, it is necessary to find new substances that can be used in treatment, either alone or as sensitizing molecules in combination with existing drugs. Peptaibols are bioactive, membrane-active peptides of non-ribosomal origin, mainly produced by filamentous fungi such as Trichoderma spp. This study focused on producing peptaibol-rich extracts from Trichoderma atroviride O1, cultivated on malt extract agar (MA) under circadian and constant darkness conditions for 13 days. Peptaibol production was detected by MALDI-TOF mass spectrometry after six days of cultivation. The extracts demonstrated antibacterial activity against Staphylococcus aureus strains, particularly the methicillin-resistant variant, but not against the Gram-negative Pseudomonas aeruginosa. Quorum sensing interference revealed that a peptaibol-rich extract suppressed Vibrio campbellii BAA-1119's AI-2 signaling system to a degree comparable with gentamycin. Beyond antibacterial properties, the extracts exhibited notable antiproliferative activity against human ovarian cancer cells and their adriamycin-resistant subline in both 2D and 3D models. Specifically, MA-derived extracts reduced ovarian cancer cell viability by 70% at 50 μg/mL, especially under light/dark regime of cultivation. Compared to previously published results for PDA-based extracts, MA cultivation shifted the biological effects of peptaibol-containing extracts toward anticancer potential. These findings support the idea that modifying fungal cultivation parameters, the bioactivity of secondary metabolite mixtures can be tailored for specific therapeutic applications.
- MeSH
- Agar * chemistry MeSH
- Anti-Bacterial Agents * pharmacology metabolism MeSH
- Hypocreales MeSH
- Culture Media chemistry MeSH
- Humans MeSH
- Microbial Sensitivity Tests MeSH
- Cell Line, Tumor MeSH
- Peptaibols * pharmacology metabolism biosynthesis chemistry MeSH
- Cell Proliferation drug effects MeSH
- Antineoplastic Agents * pharmacology metabolism MeSH
- Pseudomonas aeruginosa drug effects MeSH
- Staphylococcus aureus drug effects MeSH
- Trichoderma * metabolism growth & development chemistry MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
BACKGROUND: This study evaluated in vitro antigiardial activity in four Indonesian plants (Archidendron fagifolium, Diospyros sumatrana, Piper betle and Shorea sumatrana) extracted in methanol, methanol-tetrahydrofuran, and water. These plants exhibiting promising anti-parasitic activity were selected on the basis of collected behavioral data and their ability to decrease parasite load in Sumatran orangutans. Extracts of Arabidopsis thaliana, a plant routinely used as a laboratory model in research, were used as a negative control. METHODS: Plant extracts of different concentrations (400, 100, 25 and 6.25 μg/ml) and metronidazole (100 μg/ml), a standard giardicidal drug, were incubated with 25,000 trophozoites per milliliter of growth medium in 12-well tissue culture plates under anaerobic conditions for 72 h. Cultures were counted in a hemocytometer using a light microscope and then statistically evaluated. The cytotoxicity of the extracts was determined by the MTT Cell Viability Assay. RESULTS: We recorded a statistically-significant decrease in giardia trophozoites in two extracts: the methanolic extract of A. fagifolium (Kruskal-Wallis: p = 0.013) and the aqueous extract of P. betle (Kruskal-Wallis: p = 0.002). Effective concentrations of 400 μg/ml and 100 μg/ml, respectively, were revealed. Cytotoxicity was not demonstrated. CONCLUSIONS: The results of our study show that extracts of A. fagifolium and P. betle have potential as an alternative treatment of G. intestinalis infection. This is the first scientific proof of the biological activity of A. fagifolium, confirming the assumption that orangutans use this plant (as well as P. betle) for self-medication.
- MeSH
- Antiprotozoal Agents pharmacology MeSH
- Giardia lamblia * drug effects MeSH
- Plant Extracts * pharmacology MeSH
- Trophozoites drug effects MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Indonesia MeSH
Polymicrobial biofilms, the reason for most chronic wound infections, play a significant role in increasing antibiotic resistance. The in vivo effectiveness of the new anti-biofilm therapy is conditioned by the profound evaluation using appropriate in vitro biofilm models. Since nutrient availability is crucial for in vitro biofilm formation, this study is focused on the impact of four selected cultivation media on the properties of methicillin-resistant Staphylococcus aureus and Candida albicans dual-species biofilms. To reflect the wound environment, Tryptic soy broth, RPMI 1640 with and without glucose, and Lubbock medium were supplemented with different amounts of host effector molecules present in human plasma or sheep red blood cells. The study demonstrates that the Lubbock medium provided the most appropriate amount of nutrients regarding the biomass structure and the highest degree of tolerance to selected antimicrobials with the evident contribution of the biofilm matrix. Our results allow the rational employment of nutrition conditions within methicillin-resistant Staphylococcus aureus and Candida albicans dual-species biofilm formation in vitro for preclinical research. Additionally, one of the potential targets of a complex antibiofilm strategy, carbohydrates, was revealed since they are prevailing molecules in the matrices regardless of the cultivation media.
- MeSH
- Anti-Bacterial Agents pharmacology MeSH
- Biofilms * drug effects growth & development MeSH
- Candida albicans * drug effects physiology MeSH
- Culture Media * pharmacology MeSH
- Humans MeSH
- Methicillin-Resistant Staphylococcus aureus * drug effects physiology MeSH
- Microbial Sensitivity Tests MeSH
- Sheep MeSH
- Nutrients metabolism MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
