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Biosurfactants, sustainable alternatives to petrochemical surfactants, are gaining attention for their potential in medical applications. This study focuses on producing, purifying, and characterizing a glycolipid biosurfactant from Candida sp. UFSJ7A, particularly for its application in biofilm prevention on siliconized latex catheter surfaces. The glycolipid was extracted and characterized, revealing a critical micellar concentration (CMC) of 0.98 mg/mL, indicating its efficiency at low concentrations. Its composition, confirmed through Fourier transform infrared spectroscopy (FT-IR) and thin layer chromatography (TLC), identified it as an anionic biosurfactant with a significant ionic charge of -14.8 mV. This anionic nature contributes to its biofilm prevention capabilities. The glycolipid showed a high emulsification index (E 24 ) for toluene, gasoline, and soy oil and maintained stability under various pH and temperature conditions. Notably, its anti-adhesion activity against biofilms formed by Escherichia coli, Enterococcus faecalis, and Candida albicans was substantial. When siliconized latex catheter surfaces were preconditioned with 2 mg/mL of the glycolipid, biofilm formation was reduced by up to 97% for E. coli and C. albicans and 57% for E. faecalis. These results are particularly significant when compared to the efficacy of conventional surfactants like SDS, especially for E. coli and C. albicans. This study highlights glycolipids' potential as a biotechnological tool in reducing biofilm-associated infections on medical devices, demonstrating their promising applicability in healthcare settings., (© 2024. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Titanium implants are subject to bacterial adhesion and peri-implantitis induction, and biosurfactants bring a new alternative to the fight against infections. This work aimed to produce and characterize the biosurfactant from Bacillus subtilis ATCC 19,659, its anti-adhesion and antimicrobial activity, and cell viability. Anti-adhesion studies were carried out against Streptococcus sanguinis, Staphylococcus aureus, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and Proteus mirabilis as the minimum inhibitory concentration and the minimum bactericidal concentration. Cell viability was measured against osteoblast and fibroblast cells. The biosurfactant was classified as lipopeptide, with critical micelle concentration at 40 µg mL - 1 , and made the titanium surface less hydrophobic. The anti-adhesion effect was observed for Staphylococcus aureus and Streptococcus sanguinis with 54% growth inhibition and presented a minimum inhibitory concentration of 15.7 µg mL - 1 for Streptococcus sanguinis and Aggregatibacter actinomycetemcomitans. The lipopeptide had no cytotoxic effect and demonstrated high potential application against bacterial biofilms., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Phytases [myo-inositol(1,2,3,4,5,6) hexakisphosphate phosphohydrolases] are phytate-specific phosphatases not present in monogastric animals. Nevertheless, they are an essential supplement to feeding such animals and for human special diets. It is crucial, hence, the biotechnological use of phytases with intrinsic stability and activity at the acid pHs from gastric environments. Here we use Metadynamics (METADY) simulations to probe the conformational space of the Aspergillus nidulans phytase and the differential effects of pH and glycosylation in this same space. The results suggest that strategic combinations of pH and glycosylation affect the stability of native-like conformations and alternate these structures from a metastable to a stable profile. Furthermore, the protein segments previously reported as more thermosensitive in phytases from this family present a pivotal role in the conformational changes at different conditions, especially H2, H5-7, L8, L10, L12, and L17. Also, the glycosylations and the pH-dependent charge balance modulate the mobility and interactions at these same regions, with consequences for the surface solvation and active site exposition. Finally, although the glycosylations have stabilized the native structure and improved the substrate docking at all the studied pHs, the data suggest a higher phytate receptivity at catalytic poses for the unglycosylated structure at pH 6.5 and the glycosylated one at pH 4.5. This behavior agrees with the exact change in optimum pH reported for this enzyme, expressed on low or high glycosylating systems. We hope the results and insights presented here will be helpful in future approaches for rational engineering of technologically promising phytases and intelligent planning of their heterologous expression systems and conditions for use.Communicated by Ramaswamy H. Sarma.

Background: Due to the importance of not mistaking when determining the brain death (BD) diagnostic, reliable confirmatory exams should be performed to enhance its security. This study aims to evaluate the intracranial pressure (ICP) pulse morphology behavior in brain-dead patients through a noninvasive monitoring system., Methods: A pilot case-control study was conducted in adults that met the BD national protocol criteria. Quantitative parameters from the ICP waveforms, such as the P2/P1 ratio, time-to-peak (TTP) and pulse amplitude (AMP) were extracted and analyzed comparing BD patients and health subjects., Results: Fifteen patients were included, and 6172 waveforms were analyzed. ICP waveforms presented substantial differences amidst BD patients when compared to the control group, especially AMP, which had lower values in patients diagnosed with BD ( p < 0.0001) and the TTP median ( p < 0.00001), but no significance was found for the P2/P1 ratio ( p = 0.8). The area under curve for combination of parameters on the BD prediction was 0.77., Conclusions: In this exploratory study, noninvasive ICP waveforms have shown potential as a screening method in patients with suspected brain death. Future studies should be carried out in a larger population.

Background: Biosurfactants are natural bioactive compounds produced from fungi, bacteria and plants. These molecules have several properties that enable them to be involved in various industrial applications. The surface-active properties of biosurfactants allow their use in various sectors, such as agriculture, health, cosmetics, bioremediation/ petroleum, mining, and bioprocess., Objective: The objective of this study was to analyze the patents deposited in intellectual property databases for applications of biosurfactants in the areas of agriculture, health, cosmetics, bioremediation/petroleum, mining, and bioprocesses, with the research period consisting of the last six years., Methods: This study targeted the use of biosurfactants in various industrial sectors. The patent search was carried out using the Google Patents platform, a platform that is commonly used for this purpose and freely accessible., Results: During the search for patents related to applications of biosurfactants in industry filed in the last 6 years, results were found to be distributed in the following areas: agriculture (70), cosmetics (2311), bioremediation/petroleum (179), health (1794), mining (5), and bioprocess (133). Six patents were selected from each area, except for health and mining, for which the search resulted in only 5 and 4 patents, respectively, to be discussed and provide information on the biotechnological applications of biosurfactants in the industry., Conclusion: The present study showed a wide area of application of biosurfactants in industry. The interest in the inclusion of biosurfactants in the industry is directly related to the need for more sustainable solutions to solve real market problems. The cosmetics sector presented the most patents that employ the use of biosurfactants., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Background: Botulism is a rare and potentially fatal neuroparalytic syndrome caused by the gram-positive anaerobe spore-forming bacterium Clostridium botulinum . The microorganism produces a neurotoxin that inhibits the presynaptic release of acetylcholine at the neuromuscular junction, clinically leading to a myasthenic syndrome., Objective: To describe the recent outbreak of botulism cases and its demographic, clinical, and laboratory characteristics., Methods: We report 4 patients with botulism in the recent outbreak occurred between 2017 and 2019 in the state of Amazon., Results: Out of four patients with botulism, three contracted it from eating contaminated food and one had wound botulism. We emphasize the excellent clinical outcome of the different disease presentations in our case series., Conclusion: The temporal proximity of these reports may suggest a new rise in the number of cases in the upcoming years. A possible hypothesis is that the rarity of the disease decreased the awareness regarding the primary prevention or even a diagnosis by an untrained physician., Competing Interests: The authors have no conflict of interests to declare., (Academia Brasileira de Neurologia. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/).)

Creutzfeldt-Jakob disease (CJD) is a rare and fatal condition that leads to progressive neurodegeneration due to gliosis, vacuolation of central nervous system tissue, and loss of neurons. Microglia play a crucial role in maintaining Central Nervous System (CNS) homoeostasis, both in health and disease, through phagocytosis and cytokine production. In the context of CJD, the immunomodulatory function of microglia turns it into a cell of particular interest. Microglia would be activated by infectious prion proteins, initially acquiring a phagocytic and anti-inflammatory profile (M2), and producing cytokines such as IL-4, IL-10, and TGF-β. Therefore, microglia are seen as a key target for the development of new treatment approaches, with many emerging strategies to guide it towards a beneficial role upon neuroinflammation, by manipulating its metabolic pathways. In such a setting, many cellular targets in microglia that can be involved in phenotype modulation, such as membrane receptors, have been identified and pointed out as possible targets for further experiments and therapeutic approaches. In this article, we review the major findings about the role of microglia in CJD, including its relationship to some risk factors associated with the development of the disease. Furthermore, considering its central role in neural immunity, we explore microglial connection with other elements of the immune system and cell signalling, such as inflammasomes, the complement and purinergic systems, and the latest finding strategies to guide these cells from harmful to beneficial roles.

Amazonian palm berries (açaí, Euterpe oleracea Mart.) are fruits with high nutritional value and antioxidant activity and have aroused the interest of consumers, popularizing fruit pulps enriched with probiotics. Amazonian palm berries (açaí, Euterpe oleracea Mart.) are fruits with high nutritional potential, providing a source of carbohydrates, fibers, proteins, lipids, vitamins, and minerals. Furthermore, açai provides several health benefits, including antioxidant activity. Nutritionally enhanced foods have aroused the interest of consumers, popularizing fruit pulps enriched with probiotics. Probiotics are dietary supplements consisting of live, beneficial microorganisms in the host which improve the intestinal microbiota. The objective of this study was to isolate, identify, and characterize the probiotic potential of an isolated Schleiferilactobacillus harbinensis strain (dubbed Ca12) and provide an optimized bioprocess for its production, using the complete factorial and central rotational compound design to supplement the frozen açai pulp. The isolated strain S. harbinensis Ca12 presented adequate resistance to gastric juice and bile salts, microbial activity against different Candida strains, self-aggregation and coaggregation properties, high adhesion in HT-29 cells, and 35% inhibition of Salmonella in HT-29 cells. When optimized, the cellular biomass production of the S. harbinensis Ca12 strain was approximately 600% higher than the unsupplemented whey, with a production of 3.6 × 1010 CFU mL -1 . The S. harbinensis Ca12 strain's viability in the creamy and traditional frozen açai pulp was shown to be stable for up to 6 months at 20 °C. The impact of this study involved for the first time the S. harbinensis Ca12 described in the Brazilian cocoa pulp with activity against Candida albicans of clinical importance, creating the potential of a new functional food with important benefits to human health as prevention for candidiasis., (© 2021. Sociedade Brasileira de Microbiologia.)

Background: Highly performed nowadays, the pterional craniotomy (PC) has several widespread variants. However, these procedures are associated with complications such as temporalis muscle atrophy, facial nerve frontal branch damage, and masticatory difficulties. The postoperative cranial aesthetic is, nonetheless, the main setback according to patients. This review aims to map different pterional approaches focusing on final aesthetics., Methods: This review follows the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement. Studies were classified through the Oxford method. We searched PubMed/MEDLINE, EMBASE, and Cochrane Library from January 1969 to February 2021 for cohorts and randomized clinical trials that met our inclusion criteria., Results: 1484 articles were initially retrieved from the databases. 1328 articles did not fit the inclusion criteria. 118 duplicates were found. 38 studies were found eligible for the established criteria. 27 (71.05%) were retrospective cohorts, with low evidence level. Only 5 (13.15%) clinical trials were found eligible to the criteria. The majority of the studies (36/38) had the 2B OXFORD evidence level. A limited number of studies addressed cosmetic outcomes and patient satisfaction. The temporal muscle atrophy or temporal hollowing seems to be the patient's main complaint. Only 17 (44.73%) studies addressed patient satisfaction regarding the aesthetics, and only 10 (26.31%) of the studies reported the cosmetic outcome as a primary outcome. Nevertheless, minimally invasive approaches appear to overcome most cosmetic complaints and should be performed whenever possible., Conclusion: There are several variants of the classic PC. The esthetic outcomes are poorly evaluated. The majority of the studies were low evidence articles., Competing Interests: There are no conflicts of interest., (Copyright: © 2021 Surgical Neurology International.)

Background: Traumatic brain injury (TBI) represents one of the leading public health problems and a significant cause of neurological damage. Unintentional causes of TBI are the most frequent. However, fruit falling over the head causing TBI is extremely rare. In the Amazon region, accidents with ouriços, a coconut-like shell fruit, seem relatively common. However, to the best our knowledge, it has never been described in a scientific journal before. Therefore, we aim to evaluate a series of TBI caused by this tropical fruit., Methods: This study is a retrospective review of 7 TBI cases due to the fall of ouriços admitted to two tertiary hospitals in the Amazon region from January 2017 to December 2018. The collected data included: age, Glasgow Coma Scale, skull fracture, venous sinus injury, hematoma, surgical treatment, and outcome., Results: All patients were men, with an average age of 38, ranging from 8 to 77-years-old. Four out of seven had skull fractures. Five patients developed an epidural hematoma, and two of them had an associated subdural hematoma. Dura mater injury was observed in two patients, whereas four patients were operated. There was one related death., Conclusion: This case series is the first to describe an unconventional but potentially fatal cause of TBI in the Amazon: the falling of the Brazil-nut fruit. Most patients were diagnosed with mild TBI. Nevertheless, patients may have cranial fractures and epidural hematomas, leading to death when there's a delay in medical assistance., Competing Interests: There are no conflicts of interest., (Copyright: © 2021 Surgical Neurology International.)

The inactivation of bacteria Escherichia coli (E. coli) by non-thermal plasma (NTP) was investigated using argon, air and 1:1 mixture of air/Ar as plasma gas on five different reactors. The experiments were carried out in triplicate in each reactor, using 60 mL of distilled water pre-inoculated with E.coli. The physical-chemical analysis of pH, conductivity, nitrite, nitrate and temperature were performed soon after of 10 min of NTP treatment. The microbiological analysis of E. coli inactivation was performed using 100 μL samples withdrawn from the plasma reactor after 10 min and compared with the positive and negative control test results. The best performance were achieved whit the NTP reactors working with the upper electrode in the gas phase using 1:1 air/Ar and air as plasma gas. The results are linked with the E. coli inactivation due to membrane rupture by the NTP discharge followed by the attack of the reactive species produced in the solution. The E. coli inactivation was only partial using argon as plasma gas and the direct barrier discharge reactors showed partial inactivation even when air was used as plasma gas., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Background: Diagnosing brain death (BD) with accuracy and urgency is of great importance because an early diagnosis may guide the clinical management, optimize hospital beds, and facilitate organ transplantation. The clinical diagnosis of nonreactive and irreversible coma can be confirmed with additional tests. Among the complimentary exams that may testify brain circulatory arrest, transcranial Doppler (TCD) can be an option. It is a real-time, bedside, inexpensive, noninvasive method that assesses cerebral blood flow. In patients with suspected BD, especially those who are under sedative drugs, early diagnosis is imperative. The aim of the present study was to evaluate the role of TCD in predicting BD., Methods: One hundred consecutive comatose patients with a Glasgow Coma Scale score of less than 5, owing to different etiologies, were included. TCD was performed in all patients. The TCD operator was blinded for clinical and neurologic data. This study is in compliance with the Declaration of Helsinki., Results: Sixty-nine patients with TCD-brain circulatory collapse were diagnosed later with BD. Of the 31 patients with brain circulatory activity, 8 (25.8%) were clinically brain dead and 23 (74.2%) were alive. TCD showing brain circulatory collapse had a sensitivity of 89.6%, specificity of 100%, positive predictive value of 100%, and negative predictive value of 74.2%., Conclusion: TCD is highly specific (100%) and sensitive (89.6%) as a method to confirm the clinical diagnosis of BD, even in patients under sedation. The possibility of patients presenting with cerebral circulatory activity and clinical diagnosis of BD was demonstrated to occur., (Copyright © 2021 Elsevier Inc. All rights reserved.)

The citrus industry has grown exponentially as a result of increasing demand on its consumption, giving it high standing among other fruit crops. Therefore, the citrus sector seeks rapid, easy, and non-destructive approaches to evaluate in real time and in situ the external and internal changes in physical and nutritional quality at any stage of fruit development or storage. In particular, vitamin C is among the most important micronutrients for consumers, but its measurement relies on laborious analytical methodologies. In this study, a portable near infrared spectroscopy (NIRS) sensor was used in combination with chemometrics to develop robust and accurate models to study the ripeness of several citrus fruits (oranges, lemons, clementines, tangerines, and Tahiti limes) and their vitamin C content. Ascorbic acid, dehydroascorbic acid, and total vitamin C were determined by HILIC-HPLC-UV, while soluble solids and total acidity were evaluated by standard analytical procedures. Partial least squares regression (PLSR) was used to build regression models which revealed suitable performance regarding the prediction of quality and ripeness parameters in all tested fruits. Models for ascorbic acid, dehydroascorbic acid, total vitamin C, soluble solids, total acidity, and juiciness showed Rcv2 = 0.77-0.87, Rcv2 = 0.29-0.79, Rcv2 = 0.77-0.86, Rcv2 = 0.75-0.97, Rcv2 = 0.24-0.92, and Rcv2 = 0.38-0.75, respectively. Prediction models of oranges and Tahiti limes showed good to excellent performance regarding all tested conditions. The resulting models confirmed that NIRS technology is a time- and cost-effective approach for predicting citrus fruit quality, which can easily be used by the various stakeholders from the citrus industry., (© AOAC INTERNATIONAL 2020. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Fish fraud is a problematic issue for the industry. For it to be properly addressed will require the use of accurate, rapid, and cost-effective tools. In this work, near infrared reflectance spectroscopy (NIRS) was used to predict nutritional values (protein, lipids, and moisture) as well as to discriminate between sources (farmed vs. wild fish) and conditions (fresh or defrosted fish). Samples of five whitefish species-Alaskan pollock (Gadus chalcogrammu), Atlantic cod (G. morhua), European plaice (Pleuronectes platessa), common sole (Solea solea), and turbot (Psetta maxima)-including farmed, wild, fresh, and frozen ones, were scanned by a low-cost handheld near infrared reflectance spectrometer with a spectral range between 900 and 1700 nm. Several machine learning algorithms were explored for both regression and classification tasks, achieving precisions and coefficients of determination higher than 88% and 0.78, respectively. Principal component analysis (PCA) was used to cluster samples according to classes where good linear discriminations were denoted. Loadings from PCA revealed bands at 1150, 1200, and 1400 nm as the most discriminative spectral regions regarding classification of both source and condition, suggesting the absorbance of OH, CH, CH2, and CH3 groups as the most important ones. This study shows the use of NIRS and both linear and non-linear learners as a suitable strategy to address fish fraud and fish QC., (© AOAC INTERNATIONAL 2020. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Background: For screening probiotic strains with viability and stability in non-dairy foods for health benefits, we revised all patents relating to probiotics in food., Objective: Screening of potential probiotics from Brazilian Minas artisanal cheese and verify their survival in frozen Brazilian cocoa pulp., Methods: Isolation and identification of the strains. The potential probiotic characterization involved gastric juice and bile resistance, antibiotic and antimicrobial activity, hydrophobicity, autoaggregation, coaggregation and adhesion assay in HT-29 cells. Organoleptic, viability and stability of probiotic strain in frozen cocoa pulp were evaluated., Results: Fourteen strains of Lactobacillus plantarum (9), Weissella paramesenteroides (3), Lactobacillus fermentum (1), and Leuconostoc mesenteroides (1) were obtained. Most of the strains were resistant to simulated gastric acidity and bile salts. Almost all strains were sensitive to the antibiotics tested, except to ciprofloxacin and vancomycin. About 47% of the strains are potential producers of bacteriocins. High hydrophobicity was observed for four strains. Autoaggregation ranged from 8.3-72.6% and the coaggregation capacity from 5.2-60.2%. All of the assessed strains presented more than 90% of adhesion to HT-29 intestinal cells. The percentage of Salmonella inhibition in HT-29 cells ranged from 4.7-31.1%. No changes in color, aroma, and pH were observed in cocoa pulps after storage at -20 °C for 90 days., Conclusion: Wild strains of acid lactic bacteria from cheese proved to be viable and stable in frozen Brazilian cocoa pulp. This work showed a promising application of L. plantarum isolated strains to be used with frozen cocoa pulp matrix in probiotics food industry., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Background: Numerous causes of infection in arthroplasties are related to biofilm formation on implant surfaces. In order to circumvent this problem, new alternatives to prevent bacterial adhesion biosurfactants-based are emerging due to low toxicity, biodegradability and antimicrobial activity of several biosurfactants. We revised all patents relating to biosurfactants of applicability in orthopedic implants., Methods: This work aims to evaluate the capability of a lipopeptide produced by Bacillus subtilis ATCC 19659 isolates acting as inhibitors of the adhesion of Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 29213 onto titanium and stainless steel surfaces and its antimicrobial activity., Results: The adhesion of the strains to the stainless-steel surface was higher than that of titanium. Preconditioning of titanium and stainless-steel surfaces with 10 mg mL-1 lipopeptide reduced the adhesion of E. coli by up to 93% and the adhesion of S. aureus by up to 99.9%, suggesting the strong potential of lipopeptides in the control of orthopedic infections. The minimal inhibitory concentration and minimum bactericidal concentration were 10 and 240 µg mL-1 for E. coli and S. aureus, respectively., Conclusion: The lipopeptide produced by Bacillus subtilis ATCC 19659 presented high biotechnological application in human health against orthopedic implants infections., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Background: Phytases are enzymes capable of degrading phytic acid and used in animal feed supplementation in order to improve digestibility through the release of minerals such as phosphorus., Objective: The main goal of this study was to express and characterize a Yersinia intermedia phytase expressed in Escherichia coli cells., Methods: The Y. intermedia phytase gene was synthesized and overexpressed in Escherichia coli cells. The phytase recombinante (rPHY) was purified to homogeneity using a Ni-NTA column. The biochemical and biophysical properties of the rPHY were measured in order to fully characterize the recombinant enzyme. The following patents database were consulted: Espacenet, USPTO, LATIPAT, Patent Scope, WIPO and Google Patents., Results: The results showed that the rPHY is active at 37-40ºC and presented an optimal pH and temperature of 8.0 and 40°C, respectively. The phytase rPHY was activated by Cu2+ ion and showed resistance to trypsin and pepsin, retaining 55% of the activity at the ratio of 0.02. Furthermore, the dissociation constant (Kd = 1.1150 ± 0.0087 mM), as estimated by a fluorescence binding assay, suggests a medium affinity of the enzyme with the substrate., Conclusion: The results of this article can be considered as innovative and for this reason, they were protected by Intellectual Property Law in Brazil. Take together, the biochemical properties of the rPHY could be useful in future for its industrial application of this enzyme as an additive in the monogastric feed., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Yeasts capable of growth on xylose were isolated from macaúba (Acrocomia aculeata) fruit, a Brazilian palm tree with great potential for use as biodiesel feedstock production. Candida boidinii UFMG14 strain achieved the highest ethanol production (5 g/L) and was chosen to ferment macaúba presscake hemicellulosic hydrolysate (MPHH). The MPHH was produced by the first time in this work and the resultant fivefold concentrate showed considerable sugar content (52.3 and 34.2 g/L xylose and glucose, respectively) and low furfural (0.01 g/L) and hydroxymethylfurfural (0.15 g/L) concentrations. C. boidinii UFMG14 fermentation was evaluated in supplemented and non-supplemented MPHH containing either 10 or 25 g/L of xylose. The maximum ethanol production (12 g/L) was observed after 48 h of fermentation. The ethanol yield was significantly affected by supplementation and concentration of MPHH while ethanol productivity was affected only by MPHH concentration. This is the first study demonstrating theC. boidinii potential for ethanol production from hemicellulose byproducts., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Inactivation of the pgg2 gene, a polygalacturonase-encoding gene from Penicillium griseoroseum, reduced the total activity of polygalacturonase (PG) by 90 % in wild-type P. griseoroseum, which indicates that the pgg2 gene is the major gene responsible for PG production in this species. To increase PG production, the coding region of the pgg2 gene was cloned under the control of the glyceraldehyde 3-phosphate dehydrogenase (gpd) promoter and the terminator region of the tryptophan synthase (trpC) gene from Aspergillus nidulans (pAN52pgg2 vector). This vector was then used to transform P. griseoroseum. The transformed strains were characterized according to PG production using glucose, sucrose, or sugar cane juice as the carbon sources. The recombinant P. griseoroseum T146 strain contained an additional copy of the pgg2 gene, which resulted in a 12-fold increase in PG activity when compared with that detected in the supernatant of the control PG63 strain. The proteins secreted by the recombinant strain T146 showed a strong band at 38 kDa, which corresponds to the molecular weight of PG of the P. griseoroseum. The results demonstrate the significant biotechnological potential of recombinant P. griseoroseum T146 for use in PG production.

Background: The microbial bioemulsifiers was surface active compounds, are more effective in stabilizing oil-in-water emulsions. The yeasts have been isolated to produce bioemulsifiers from vegetable oils and industrial wastes., Results: Trichosporon mycotoxinivorans CLA2 is bioemulsifier-producing yeast strain isolated from effluents of the dairy industry, with ability to emulsify different hydrophobic substrates. Bioemulsifier production (mg/L) and the emulsifying activity (E24) of this strain were optimized by response surface methodology using mineral minimal medium containing refinery waste as the carbon source, which consisted of diatomaceous earth impregnated with esters from filters used in biodiesel purification. The highest bioemulsifier production occurred in mineral minimal medium containing 75 g/L biodiesel residue and 5 g/L ammonium sulfate. The highest emulsifying activity was obtained in medium containing 58 g/L biodiesel refinery residue and 4.6 g/L ammonium sulfate, and under these conditions, the model estimated an emulsifying activity of 85%. Gas chromatography and mass spectrometry analysis suggested a bioemulsifier molecule consisting of monosaccharides, predominantly xylose and mannose, and a long chain aliphatic groups composed of octadecanoic acid and hexadecanoic acid at concentrations of 48.01% and 43.16%, respectively. The carbohydrate composition as determined by GC-MS of their alditol acetate derivatives showed a larger ratio of xylose (49.27%), mannose (39.91%), and glucose (10.81%). 1 H NMR spectra confirmed by COSY suggested high molecular weight, polymeric pattern, presence of monosaccharide's and long chain aliphatic groups in the bioemulsifier molecule., Conclusions: The biodiesel residue is an economical substrate, therefore seems to be very promising for the low-cost production of active emulsifiers in the emulsification of aromatics, aliphatic hydrocarbons, and kerosene.

Aims: To obtain recombinant strains of Penicillium griseoroseum that produce high levels of pectin lyase (PL) and polygalacturonase (PG) simultaneously., Methods and Results: A strain with high production of PL was transformed with the plasmid pAN52pgg2, containing the gene encoding PG of P. griseoroseum, under control of the gpd promoter gene from Aspergillus nidulans. Southern blot analysis demonstrated that all strain had at least one copy of pAN52pgg2 integrated into the genome. The recombinant strain P. griseoroseum T20 produced levels of PL and PG that were 266- and 27-fold greater, respectively, than the wild-type strain. Furthermore, the extracellular protein profile of recombinant T20 showed two protein bands of c. 36 and 38 kDa, associated with PL and PG, respectively., Conclusions: This recombinant strain T20 produces PL and PG using carbon sources of low costs, and an enzyme preparation that is free of cellulolytic and proteolytic activities., Significance and Impact of the Study: PL and PG play an important role in the degradation of pectin. Owing to their use in the juice and wines industries, there is a growing interest in the inexpensive production of these enzymes. This work describes an efficient system of protein expression and secretion using the fungus P. griseoroseum., (© 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.)

Previous reports have described pgg2, a polygalacturonase-encoding gene of Penicillium griseoroseum, as an attractive model for transcriptional regulation studies, due to its high expression throughout several in vitro growth conditions, even in the presence of non-inducing sugars such as sucrose. A search for regulatory motifs in the 5' upstream regulatory sequence of pgg2 identified a putative CCAAT box that could justify this expression profile. This element, located 270 bp upstream of the translational start codon, was tested as binding target for regulatory proteins. Analysis of a 170 bp promoter fragment by electrophoretic mobility shift assay (EMSA) with nuclear extracts prepared from mycelia grown in pectin-containing culture medium revealed a high mobility complex that was subsequently confirmed by analyzing it with a double-stranded oligonucleotide spanning the CCAAT motif. A substitution in the core sequence for GTAGG partially abolished the formation of specific complexes, showing the involvement of the CCAAT box in the regulation of the polygalacturonase gene studied.