bluuebunny/crossref_metadata_2025_split huggingface dataset:
Utilization of orange pulp and corn steep liquor for L-methioninase production by Wickerhamomyces subpelliculosus
Authors: Hassabo Amany A. , Mostafa Elsayed E. , Saad Moataza M. , Selim Mohsen H. 
Published: August 2024

Background and objective
L-methioninase has attracted much attention with respect to its proposed applications in both pharmaceuticals and food industry. The aim of this study was to develop an economic medium formulation using agro-industrial by-products as substrates for large-scale production of L-methioninase.

Materials and methods
Identification of a high L-methioninase-producing yeast isolate was carried out using 18S rRNA molecular technique. Screening of various agro-industrial by-products and optimization of different process parameters were investigated. Partial purification and characterization of a crude enzyme were carried out.

Results and discussion
A high L-methioninase-producing yeast isolate was phylogenetically identified as Wickerhamomyces subpelliculosus. Among different agro-industrial by-products tested, orange pulp supported maximum enzyme production (94.08 U/ml) followed by cane and beet molasses. In addition, corn steep liquor (CSL) gave high enzyme level (141.12 U/ml) and could be used as an inexpensive alternate for yeast extract. The optimum growth conditions were found to be orange pulp 30% (w/v), CSL 4% (v/v), CaCl2 0.05%, and KH2PO4 0.05% (w/v) at pH 6.0 after 48 h of incubation. This developed medium formulation increased L-methioninase production (161.95 U/ml) by twofold compared with that obtained by the Czapek–Dox’s medium (73.92 U/ml). Crude enzyme was partially purified by heat treatment at 70°C with 2.9 purification fold. The enzyme activity was optimal at temperature 60°C and pH 7.0. The results showed that a mixed formulation of orange pulp and CSL can be used as an effective and economic substrate for the production of L-methioninase by W. subpelliculosus.
Statistical optimization of lipase production in solid-state fermentation by Aspergillus tamarii NDA03a and application of the fermented solid as a biocatalyst for biodiesel production
Authors: Ahmed Hanan M. , Mohamed Sayeda S. , Moharam Maysa E. , El-bendary Magda A. , Abd El-lateaf Hisham A. , Amin Hala A. 
Published: August 2024

Background and objective
Biodiesel, an attractive alternative fuel, is defined by the American Society for Testing and Materials (ASTM) as fatty acid methyl esters (FAME). Biodiesel is an ecofriendly fuel compared with many other transportation fuels. The aim of this study was to implement the statistical approaches for optimization of Aspergillus tamarii NDA03a mutant G lipase produced in solid-state fermentation (SSF), and then application of the dried fermented solid as a biocatalyst for biodiesel production from waste frying oil (WFO).

Materials and methods

A. tamarii NDA03a mutant (3G) was previously selected as a good lipase producer. Five oil residue meals were evaluated in the presence of wheat bran (WB) for their potential as enzyme inducers and substrates for the production of 3G lipase by SSF. The best oil residue meal was selected and used in subsequent experiments. The fermented solid thus obtained was collected, lyophilized, and used as a biocatalyst for waste frying oil transesterification to FAME. To optimize SSF conditions for lipase production using 3G, a Plackett–Burman design was used at first to screen the critical factors from several process variables, and finally, a central composite design was applied to further estimate the relationship between the variables and response as well as optimize the levels. Response was measured in terms of FAME yield. To verify the adequacy and accuracy of the model, validation experiments were also carried out.

Results and conclusion
The most favorable oil residue meal that enhances 3G lipase production by SSF was black cumin meal. Results of the Plackett–Burman design revealed that the factors contributing to the main effect were incubation temperature, incubation period, and moisture content. The optimal SSF conditions for lipase production were WB 10 g, black cumin meal 6% (w/w of WB), pH 8, temperature 28°C, moisture content 40%, molasses 1% (w/w of WB), and incubation period 3 days. Under these optimized conditions, produced FAME yield (65.55%) increased by 58% compared with the basal medium (41.46%). A good agreement between the experimental (65.55%) and predicted (65.03%) values was detected. The significance of this model was confirmed by its probability value and lack of fit (P<0.05) and clearly showed that the model was sufficient to describe the correlation between the FAME yield and the tested variables. The obtained results ascertained the success of response surface methodology as an efficient technique to optimize the lipase production in SSF and consequently the ability of application of the dried fermented solid as a biocatalyst for biodiesel production.
Solid-state fermentation and optimization of cellulase production using local fungal isolate
Authors: Basha Maha A. , Shetaia Yousseria M. , Mehaya Fathy M. , Abdelzaher Fatma H. 
Published: August 2024

Background
Cellulase is the most employed industrial enzyme in biological conversion of many cellulosic wastes. In this work, economic cellulase production by fungi in solid-state fermentation (SSF) by using solid wastes of medicinal plants was studied. Optimization of growth conditions for production of cellulase was the main target of this study.

Objective
The current study aimed to isolate and screen fungal isolates that have the ability to produce enzymes to degrade solid wastes of medicinal plant process and optimization of growth factors that affect cellulase production.

Materials and methods
Thirty-five fungal isolates were isolated from different sources by plating and screened for their cellulase activities using Czapek–Dox broth medium amended with 1% cellulose. Cellulase production by tested fungal isolates was carried out through utilization of olive (Olea europaea), black seeds (Nigella sativa), and castor bean (Ricinus communis) cakes in SSF. Optimization of the cellulase productivity was performed by Plackett–Burman design (PBD) and Box–Behnken design.

Results and conclusion
Out of the isolated 35 fungi, only 12 (34%) produced cellulase in SSF using olive, black seeds (Nigella), and castor bean cakes. Out of these fungal isolates, only 4, that is, no. 1, 7, 10, and 17 were superior in reducing sugar production from olive cakes (13.04, 15.61, 17.03, and 12.85 mg/ml), respectively. While four fungal isolates no. (1, 7, 7, and 10) were active producers of reducing sugars from black seeds (15.45, 18.96, 20, and 18.08 mg/ml), respectively. Only a fungal isolate no. 7 gave high reducing sugars (15.34 mg/ml) in castor cake SSF. The most potent fungal isolate (no. 10) produced 20 mg/ml of reducing sugars using black seed cakes as substrate for SSF. The potential fungal isolate was identified as Aspergillus terreus (OQ085169) based on the extracted fungal DNA that was amplified by PCR using specific internal-transcribed spacer primer (ITS1/ITS4). The PCR products were sequenced and compared with the other related sequences in GenBank (NCBI).
The screening of seven factors using PBD showed that only three variables: pH, incubation time, and aeration rate (rpm) affected significantly cellulase production. Box–Behnken design was used to estimate the optimal level of the selected variables based on the results of the PBD. All variables increased significantly cellulase using A. terreus (OQ085169). The P value was very low (0.0207) that indicated the significant, high correlation between the predicted and actual values (R
2=0.98), this indicating 98% of the variation in the cellulase activity was owing to the selected independent variables.
Analytical characterization of biomarkers in an optimized novel antidiabetic polyherbal formulation using high-performance thin-layer chromatography and liquid chromatography with tandem mass spectrometry
Authors: Jayaraju Kummu Jyothsna , Ishaq Beludari Mohammed 
Published: August 2024

Background
Diabetes mellitus is a chronic health issue that requires novel approaches to treatment and a multimodal approach to prevention. In the treatment of diabetes, a polyherbal formulation is the finest alternative medicine. A polyherbal formulation was developed in-house and evaluated for its antidiabetic potential on streptozotocin-induced diabetes rat. The same extract was now characterized analytically utilizing a variety of methods.

Objective
The goal of this study was to quantify the biomarkers in a novel antidiabetic polyherbal formulation made in-house with Cinnamonum zeylanicium bark, Eugenia jambolana seeds, Vinca rosea whole plant, and Gymnema sylvestre (GS) leaves, using high-performance thin-layer chromatography (HPTLC) and liquid chromatography with tandem mass spectrometry (LC–MS/MS).

Materials and methods
Cinnamaldehyde (CIN), gallic acid (GLA), vincristine (VC), vinblastine (VB), and gymnemic acid (GYA) were identified as bioactive components of polyherbal formulation hydroalcoholic extract utilizing HPTLC and LC–MS/MS. Acetonitrile, methanol, and 0.1 percent formic acid was used as mobile phase, chromatographic separation was accomplished in 30 min using a gradient system and a SUNFIRE C18, 250×4.6, 5-µm analytical column with a flow rate of 1.0 ml/min in LC–MS/MS research. Scanned in a positive mode with a scan speed of 100–2000 AMU/s over a mass range of 20–1974 Da. The electron-spray ionization mode was used, with a source temperature of 150°C and a desolvation temperature of 350°C. The HPTLC separation was performed using ethyl acetate/acetonitrile/water/formic acid/N-dimethyl formamide 5.5 : 2.5 : 0.5 : 1.0 : 0.5 (v/v) as the mobile phase on precoated silica gel 60 GF254 plates. At room temperature, the plates were developed to a distance of 9.0 cm. CIN, GLA, VC, VB, and GYA plates were scanned and measured at wavelengths of maximum absorption of 259, 287, 342, 355, and 387 nm, respectively. Band size, chamber-saturation duration, migration of the solvent front, slit width, and other experimental parameters were carefully examined, and the optimized chromatographic conditions were chosen.

Results
LC–MS analysis of the hydroalcoholic extract of the polyherbal formulation revealed the presence of all the five bioactive chemical constituents, CIN, GLA, VC, VB, and GYA. Similarly, the drug samples were satisfactorily resolved with Rf 1.81±0.01, 0.05±0.01, 0.02±0.01, 0.09±0.01, and 0.04±0.01 for CIN, GLA, VC, VB, and GYA respectively, using HPTLC.

Conclusion
The importance of combining Ayurvedic formulations with contemporary high-throughput screening techniques will spark new interest in more powerful biocompatible drug leads. The findings of this study lend scientific credence to the therapeutic applications of the polyherbal formulation.
Enhancement of some barley (Hordeum vulgare L.) resistance for nematode (Heterdra avanae) using DNA fingerprinting analysis
Authors: Rashad Shimaa E. , Eldemerdash Ibthal S. , Hamed Hamdy M. , El-Enany Magda A.M. , Heiba Samy A.A. 
Published: August 2024

Background

Heterodera avenae, the cereal cyst nematode or European cyst nematode, is a plant pathogen and an obligate parasite of cereal crops including barley, oats, wheat, and rye. Cereal crops infected with this nematode are more susceptible to infection by fungal diseases such as Rhizoctonia root rot.

Objective
The main objective of the present study was to map Heterodera avenae resistance and to compare seven different species. This study also aimed to increase the efficiency and precision of standard procedures for testing the resistance of barley toward root-lesion nematodes.

Materials and methods
The genetic variability analysis of seven barley genotypes, of Egypt was achieved using 7 Inter simple sequence repeat (ISSR) primers. Phylogenetic relationships of seven accessions of (Hordeum vulgare L.) collected from different region of Egypt were assessed. The soil was sterilized in an autoclave of each pot was planted in the comparison experiment, as well as in the treatment experiment with nematode Heterdra avanae infected soil brought from an infected field, and this soil was added to the treated pots. Also (ISSR) molecular marker technique was used for DNA fingerprinting and assessing genetic diversity and phylogenetic relationships in barley germplasm.

Results and conclusion
A significant correlation was observed between the Jaccard’s dissimilarity matrices based on ISSR markers, as revealed by Mantel test using the Pearson correlation coefficient (r = 0.69; P < 0.05). The results showed that ISSR primers produced 37 bands their size ranged between 100-2000 bp with (87.5%). polymorphism percentage. Polymorphic information content PIC was 0.74 for ISSR. Unweighted Pair Group Method with Arithmetic (UPGMA), Dendrogram was divided into two clusters by morphological traits and ISSR analysis. Genetic similarity matrix was examined with Jaccard’s coefficient, maximum similarity was found between Giza126 and Giza127 (99%) with morphological analysis both and lowest similarity between Giza123 and Giza126 (6%) with ISSR analysis. Determination of genetic diversity between barley is of major importance for characterization of barley germplasm, breeding programs and conservation purposes. Morphological traits and ISSR analysis are effective tools for detecting genetic variations. The results showed that H. vulgare have high ratio of variation. This study may be considered as reference study for further studies on H. vulgare and may contribute to species concept and breeding programs.
Impact of turmeric and carrot extracts on morphological, chemical composition and isozymes patterns of Azadirachta indica seedlings under water deficiency conditions
Authors: Mohamed Amr S. , El-Sayed Samah M. , Elsayed Shaimaa I.M. , Mazher Azza A.M. 
Published: August 2024

Background

Azadirachta indica trees are of great importance because of the high-quality wood they produce, which is used in a wide range of industries, and their production of insect repellent oils.

Objective
Due to these great benefits that have drawn attention to them, it was necessary to find environmentally friendly solutions to improve the ability of this plant species to grow in the event of exposure to conditions of water shortage.

Materials and methods
The plants were sprayed with Carrot extract (C) at rates of 50 and 100 ml/ l and/or Turmeric extract (T) at a rate of 20 and 40 ml/l under irrigation intervals every 4 and 8 days. Total chlorophyll content, total sugars content (mg/g F.W.), flavonoids content (mg/g F.W.), total phenols (mg/g F.W.) and total indoles (mg/100 g F.W.) were determined. Also, Peroxidase isozymes (POD) and Polyphenol oxidase isozymes (PPO) were determined.

Results and conclusion
The obtained results confirmed that increasing irrigation periods (8 days) had a negative effect on most vegetative traits except for all root characteristics, and also all chemical properties except the total indoles content and lipid peroxidation, it was also observed that the enzymatic activity of peroxidase isozymes (POD) and polyphenol oxidase isozymes (PPO) compared to plants that were watered every 4 days. The most of growth parameters, chemical composition and the activity of POD in plants irrigated every four days were improved with C treatment at 100 ml/l, while the activity of PPO increased with 100 ml/l C+40 ml/l T. On the other hand, all growth parameters, chemical compositions and POD in plants irrigated every 8 days were increased with 100 ml/l C +20 ml/l T except lipid peroxidation which gave the highest value in control plants, whereas PPO increased with C treatment 50 ml/l+T 40 ml/l.
Production of isoflavones-enriched soy yogurt through soymilk fermentation using probiotic bacteria
Authors: El-Shazly Asmaa I. , Gamal Amira A. , El-Dein Asmaa N. , Mettwally Walaa S.A. , Farid Mohamed A. 
Published: August 2024

Background and objective
Fermented soy products were regarded as healthy foods and hence are considered an essential part of the diet. Lactic acid bacteria isolated from naturally fermented Egyptian food products were screened for their ability to produce β-glucosidase, isoflavone aglycone, phenolics, and antioxidant activity during the formation of soy yogurt. The present research is a preliminary attempt to ascertain soy yogurt production by different strains of lactic acid bacteria and their efficacy for the production of the aforementioned products.

Materials and methods
A total of 16 probiotic lactic acid bacteria were used for the preparation of soy yogurt and tested for their probiotic properties. Soymilk was prepared and inoculated (1% v/v) with the probiotic strains previously activated in the MRS medium. After fermentation, cell viability, pH, titratable acidity, total phenolic compounds concentration, antioxidant activity, isoflavones aglycone (daidzein and genistein), and extracellular and cell membrane-bound β-glucosidase activity were determined.

Results and conclusion
A total of 16 probiotic lactic acid bacteria were used for the preparation of soy yogurt. The final pH of the fermented soymilk ranged from 4.92 to 6.6, and their titratable acidity (lactic acid %) ranged from 0.5 to 0.99%. Changes in β-glucosidase, isoflavone aglycone, total phenolics content, and antioxidant activity during the formation of soy yogurt were determined. All bacterial isolates showed positive cell-bound and extracellular β-glucosidase. Their activities ranged from 308.65 to 553 mU/ml. The Lactobacillus strains showed lower extracellular than their cell-bound β-glucosidase, and the opposite was true for the other group. An increase in the content of isoflavone aglycones in soy yogurt could be achieved by aging with bacterial fermentation. Soymilk fermented with Lactobacillus strains showed the highest bioconversion of isoflavone glucosides into isoflavone aglycones than other strains, although they produced less β-glucosidase enzymes. The antioxidant activity is related to changes in total phenolics. All microorganisms were able to increase the total phenols, whereas some Lactobacillus strains were unable to release more phenols compared with unfermented soymilk.
Production of a bacterial extracellular L-glutaminase possessing high antioxidant capability
Authors: El-Sousy Sara M. , Easa Saadia M. , Hassan Amira A. , Ismail Abdel-Mohsen S. 
Published: August 2024

Background and objectives
L-glutaminase has utmost practical importance in many fields, such as medicine, pharmacy, and some industries as an effective antioxidant, anticancer, flavor enhancer, and used as an analytical reagent in the determination of glutamate and glutamine. The objective of the present article was to formulate the production medium and to pinpoint the proper growth conditions for the most potent microorganism producing highly active glutaminase enzyme. The general properties of the crude enzyme preparation were determined to detect the proper conditions for enzyme activity. Under the specified conditions, the capabilities of the crude L-glutaminase preparation for antimicrobial and antioxidant activities were investigated.

Materials and methods
A total of 12 recommended microbial strains were screened for highly active L-glutaminase enzyme production. Factors influencing the production of L-glutaminase enzyme were optimized, and the important properties of the crude enzyme were pinpointed. Finally, biological activities of the crude enzyme were investigated as a preliminary index for the validity of the partially purified L-glutaminase form for medical applications.

Results and conclusion
Among all tested microorganisms, Bacillus subtilis NRRL 1315 was the most potent producer for L-glutaminase enzyme. The maximum glutaminase production was obtained after 48 h of incubation on a rotary shaker (150 rpm) with the medium containing 5 g/l glucose, 0.1 g/l sodium nitrate, and 10 g/l L-glutamine at 37°C and pH 7.5. The important properties of the crude L-glutaminase were duly pinpointed as follows: optimum enzyme protein concentration and substrate concentration were 2 mg/ml and 40 mM, respectively, and optimum reaction pH and temperature were 7.5 and 37°C, respectively. Under the specified conditions, the crude enzyme exhibited considerable 2, 2′-diphenyl-1-picrylhydrazyl radical scavenging activity.
A novel UV-spectrophotometric method as a reliable alternative for the spectrofluorimetric analysis of lucifer yellow
Authors: Monir Hany H. , El-Ashmawy Ahmed A. , Abdou Aya R. , Fayad Walid , Mahmoud Khaled M. , Emara Laila H. 
Published: August 2024

Background
Human colon adenocarcinoma cell-line (Caco-2) monolayer is a well-established ex vivo model used for estimation of drug-apparent permeability through the human intestinal barrier. Lucifer yellow (LY) is a paracellular permeability marker used generally for examining the integrity of Caco-2 monolayer using spectrofluorimetric detection. No other spectroscopic methods were reported for analysis of LY.

Objective
To introduce a novel validated simple ultraviolet (UV)-spectrophotometric method, for LY determination, as an alternative to the commonly used spectrofluorimetry, as well as to apply the developed UV method to confirm the monolayer integrity of Caco-2 cell line compared with the spectrofluorimetric detection.

Materials and methods
The measurement was carried out at a wavelength of 277 nm. The linearity range was 1.0–12.5 μg/ml with the regression coefficient (R
2)=0.9992. Validation of the method was carried out following the ICH guidelines concerning linearity, precision, accuracy, robustness, and stability and statistically compared with the conventional spectrofluorimetric one. Measurement of cultured Caco-2 cell monolayer integrity was carried out using LY employing spectrofluorimetric and UV-spectrophotometric methods.

Results and conclusion
The calculated t test and F values indicated that no significant difference existed between both methods regarding precision and accuracy. The developed method has been successfully applied for checking the monolayer integrity of Caco-2 cell line. Comparing permeability percentage results of LY, both methods resulted in LY-permissible values of 2.354 and 2.303% using spectrofluorimetric and UV-spectrophotometric methods, respectively, indicating Caco-2 cell-line monolayer integrity.