Electronic Journal of Biotechnology最新文献

Background

Soil salinization is one of the key factors restricting the production of cropland. Once rice is subjected to alkali stress at the bud burst stage, the yield will suffer irreparable serious loss. Compared with salt tolerance, studies on QTL mapping and candidate gene analysis of rice alkali tolerance are limited.

Results

In this study, we used the F_2:3 population derived from the alkali-tolerant cultivar LD21 and the alkali-sensitive cultivar WL138 to construct an alkali-tolerant DNA mixing pool, and the BSA (Bulked Segregation Analysis) method was used for re-sequencing. The main QTL qRSLB9 controlling the relative shoot length of rice under alkali stress was mapped by QTL-seq. The candidate interval was narrowed to 346.5 kb by regional linkage mapping, which containing 6 DEGs screened through transcriptome sequencing. The qRT-PCR and candidate gene sequencing showed that LOC_Os09g24260 was most likely to control relative shoot length (RSL) in rice as a major gene who encodes the WD domain, G-beta repeat domain-containing protein.

Conclusions

Based on these results, LOC_Os09g24260 was the candidate gene of qRSLB9 conferring alkalinity tolerance to rice at the bud burst stage. Our study provides valuable genetic information and materials for breeding new rice varieties with alkalinity tolerance.

How to cite: Wang J, Bian J, Liu L, et al. Screening and analysis of candidate genes conferring alkalinity tolerance in rice (Oryza sativa L.) at the bud burst stage based on QTL-seq and RNA-seq. Electron J Biotechnol 2024;71. https://doi.org/10.1016/j.ejbt.2024.07.002.

Background

Different methods for the extraction of trypsin inhibitors in beans (Phaseolus spp.) were investigated. Two randomised complete laboratory experiments were performed, one on the seeds and one on the pods. In the first, the seeds of common bean variety KIS Marcelijan, breeding line Ref_316 × 498 and runner bean variety Bonela were examined. In the second, the fresh pods of five common beans (three breeding lines, two varieties) were analysed. Four extraction methods were used, including ultrasonic-assisted extraction (UAE) for 15 and 30 min and shaking-assisted extraction for 60 and 180 min.

Results

The results showed a significant increase in trypsin inhibitor activity-related traits in UAE compared to shaking extraction, with the 15 min ultrasonic process showing better efficacy than the one with 30  min duration. In the seed experiment, the breeding line Ref_316 × 498 showed the highest Trypsin Units Inhibited (TUI) and TUI/mg sample after a 15 min UAE. In the pod experiment, the breeding line 228_4aa_ca also showed the highest TUI and TUI/mg sample after a 15 min extraction with UAE. These results underline the potential of UAE to maximise trypsin inhibitor content. In addition, remarkable correlations between TUI, TUI/mg sample and the percentage of trypsin inhibition (%TIn) were observed in both experiments.

Conclusions

These results provide valuable insights into the relationship between bean genetic resources, extraction methods and trypsin inhibitor content in bean pods and seeds and serve as a basis for refining extraction protocols. The study encourages further research on the practical implications of investigated protocols for breeding programmes and agricultural practices.

How to cite: Tavakoli Hasanaklou H, Pipan B, Meglič V, et al. Trypsin inhibitors in seeds and pods of Phaseolus vulgaris/coccineus: A comparative study of shaking and ultrasonic extraction methods. Electron J Biotechnol 2024;71. https://doi.org/10.1016/j.ejbt.2024.05.003.

Background

Using cell lines to explore the function of organic compounds is fundamental in biotechnology. Evaluating new additives intended to improve animal production is challenging due to the complexity and uncertainty of in vivo testing. This study investigated the action of a compound with antioxidant properties using cells from terrestrial (LMH cells line) and aquatic vertebrates (CHSE-214).

Results

The results of our study provide reassuring evidence of the compound’s safety for use in animal production. The compound demonstrated no adverse effects on cell viability, indicating its potential for safe application. Furthermore, the compound’s antioxidant properties were evident, with a 100% recovery in both cell lines when exposed to hydrogen peroxide 0.1 mM. It also effectively reduced cellular ageing caused by metabolic processes, as measured by the TBARS formation in both cell lines, from 5 MDA µM/mg protein to 2.5 MDA µM/mg protein when used at 0.05 or 0.5 g/L. Notably, this action did not increase cell membrane oxidation, further supporting its safety profile.

Conclusions

These findings indicate that the compound has an antioxidant effect and can be used independently or in combination with metabolic stimulants in the diets of production animals. Applying this additive and its possible synergy with other compounds could help reduce oxidative stress and improve growth in animal production. The data generated in this study provide a solid basis for designing diets incorporating this additive to observe improvements in animal production based on activity observed at the cellular level.

How to cite: Olivares- Ferretti P, Maguregui E, Chavez V, et al. Novel antioxidant additive ENTAN molecule for animal production: Evaluation at the cellular level. Electron J Biotechnol 2024;71. https://doi.org/10.1016/j.ejbt.2024.07.001.

Background

Lactobacillus plantarum can produce many secondary metabolites, some of which have antibacterial effects. This study aimed to explore the main antimicrobial metabolites of Lactobacillus plantarum LPZN19.

Results

The results of antibacterial activity after fermentation for different durations showed that the metabolites from the LPZN19 cell-free supernatant (LCFS) after 24 h had the strongest antibacterial activity, which was confirmed by the highest contents of organic acids and fatty acids in the LCFS after 24 h. Lactic acid, phenyllactic acid, malic acid, aspartic acid, dodecanoic acid and propionic acid were the main differentially abundant metabolites. LCFS was separated by semi-preparative liquid chromatography to obtain 4 antibacterial parts, mainly organic acids such as lactic acid, glycolic acid, and citric acid, and fatty acids such as stearic acid, palmitic acid, and octanoic acid. In addition, fatty glycerides and amino acids with antimicrobial activity were included.

Conclusions

Our findings indicate that the main antimicrobial metabolites of L. plantarum LPZN19 include organic acids, fatty acids, fatty glycerides and some amino acids with antimicrobial activity, which not only clarifies the main antimicrobial metabolites of L. plantarum LPZN19 but also provides an effective method for rapid screening of antimicrobial substances.

How to cite: Wang Y, Xu Y. Analysis and identification of the main antimicrobial metabolites of Lactobacillus plantarum LPZN19. Electron J Biotechnol 2024;71. https://doi.org/10.1016/j.ejbt.2024.05.005.

Background

Xylitol, a five-carbon polyalcohol, is used in the food and pharmaceutical industries and as a building block in the synthesis of high-value chemicals. It can be sustainably produced from renewable sources through xylose assimilating microbe fermentation.

Results

We screened microbial strains for xylitol production and identified Wickerhamomyces anomalus Z1 as a key xylitol producer. Utilizing lignocellulosic biomass hydrolysates for xylitol production poses challenges due to microbial sensitivity to inhibitors from biomass pre-treatment. In this study, an adaptive laboratory evolution (ALE) of W. anomalus Z1 was performed by culturing the yeast in a mineral medium supplemented with gradual increases of sugarcane bagasse hemicellulosic hydrolysate (SCHH) obtained by intensified steam explosion pretreatment. The performance of the adapted yeast, named Wickerhamomyces anomalus ALE, was assessed in comparison to the wild-type strain regarding its capacity to produce xylitol using SCHH. The evolved yeast reached a xylitol yield of 0.11 g xylitol/g xylose whereas the wild-type strain could not produce xylitol. Removing acetic acid from SCHH enhanced W. anomalus ALE performance, with optimal results at 75% hydrolyzed hemicellulose, yielding 0.44 g xylitol/g xylose and 13.41 g/L xylitol.

Conclusions

This study demonstrates the potential of W. anomalus ALE in successfully valorizing the hemicellulosic fraction of sugarcane bagasse for sustainable xylitol production.

How to cite: Bonfiglio F, Cagno M, Nuñez L, et al. Xylitol production by a Wickerhamomyces anomalus strain adapted for enhanced tolerance to sugarcane bagasse hemicellulosic hydrolysate with high content of fermentation inhibitors. Electron J Biotechnol 2024;71. https://doi.org/10.1016/j.ejbt.2024.05.004.

Background

L-asparaginase (L-ASNase) is an essential enzyme used to treat acute lymphoblastic leukemia (ALL) by depleting L-asparagine, a vital nutrient for leukemia cells. However, its clinical use is challenged by adverse effects linked to its bacterial origin and L-glutaminase (L-GLNase) co-activity. This study aims to identify fungi capable of producing L-ASNase with reduced L-GLNase co-activity.

Results

Among the fungal iolates, isolate JK12 and ChL11 showed high L-ASNase activity (34.04 ± 1.83^a U/ml and 30.84 ± 0.53^b U/ml, respectively) with reduced L-GLNase co-activity (4.95 ± 0.28^c U/ml and 4.80 ± 0.02^d U/ml, respectively). Sequencing of the internal transcribed spacer (ITS) region of these isolates identified them as Candida palmioleophila isolate JK12 (≥99% identity with Candida genus) and Trichosporon asahii isolate ChL11 (≥98% identity with Trichosporon genus). Moreover, these isolates exhibited distinct preferences for carbon (C) and nitrogen (N) sources, as well as culture conditions for L-ASNase production. C. palmioleophila isolate JK12 demonstrated the highest L-ASNase production in fructose and yeast extract (67.6 ± 0.04^a U/ml and 51.4 ± 0.04^a U/ml, respectively), following 96 h of incubation at 25°C (43.8 ± 1.22^a U/ml, 55.8 ± 0.02^a U/ml, respectively), with an agitation speed of 100 rpm (59.9 ± 0.04^a U/ml). On the other hand, T. asahii isolate ChL11 exhibited maximum L-ASNase production in sucrose and L-asparagine (64.2 ± 0.08^a U/ml and 63.6 ± 0.01^a U/ml, respectively), after 120 h of incubation at 35°C.

Conclusions

The fungal isolates T. asahii isolate ChL11 and C. palmioleophila isolate JK12 have been identified as promising L-ASNase sources of safer therapeutic prospects in cancer therapy due to the reduced GLNase co-activity.

How to cite: Sisay T, Mobegi VA, Wachira S, et al. Isolation and characterization of fungi producing L-asparaginase with reduced L-glutaminase activity from soil samples. Electron J Biotechnol 2024. https://doi.org/10.1016/j.ejbt.2024.05.002.

Background

The important challenge to the biotechnology is to find new effective fusion partners that enable to improve solubility, expression, and optimize the subsequent fine purification of the target protein.

Results

The most invariant part of the most immunogenic region of the surface virulence factor A of Streptococcus pneumoniae was selected as a model target protein, while the thermostable chemotaxis polypeptide of W-type from Thermotoga petrophila was used as a fusion partner. The genes encoding fusion variants of these proteins were constructed and cloned into a plasmid vector under the control of the strong bacteriophage T7 transcription regulatory system. Effective Escherichia coli producer strains were obtained, and optimal conditions were chosen for the production of resulting constructs. The optimal pH and temperature ranges of recombinant proteins were determined, and three-dimensional shapes of their molecules were also predicted. Methods of low-stage protein purification were improved. Some of the isolated proteins demonstrated a high level of expression, solubility and purity.

Conclusions

Novel chimeric proteins were obtained which had not previously been observed in nature in such domain combinations. It was shown that the biotechnologically valuable characteristics of the hybrid proteins were more marked when the thermal-resistant partner was combined with the N-terminus of pneumococcal protein. The principles of their low-stage purification were performed which does not require any special equipment. That is a basis for significant reduction of prices for diagnostic test-systems components and subunit vaccine production.

How to cite: Grishin DV, Sidorov NG, Parfenova OK, et al. Construction, heterological expression and a simple purification of the BP region of the pneumococcal surface protein a fused in different orientations to the chemotaxis adaptor protein CheW from Thermotoga petrophila. Electron J Biotechnol 2024;71. https://doi.org/10.1016/j.ejbt.2024.05.001.

Background

Chinese hamster ovarian (CHO) cells are widely employed in biotechnology for the production of recombinant proteins. Extending the life span of CHO cells and inhibiting the loss of producing cell population through the inhibition of apoptosis can benefit the productivity of those cells. In this study, we aimed to screen and evaluate the impact of some caspase-7 inhibitor candidates on the lifespan of CHO cells.

Results

Through virtual screening and molecular docking, risperidone was screened and selected as a potential inhibitor of caspase-7 in CHO cells. The results of MTT assay revealed that the cytotoxicity of risperidone at all concentrations was lower than 50%, and thus it can be suggested as a safe treatment for CHO cells. Annexin V apoptosis and flow cytometry assays revealed that risperidone at 1, 25, and 50 µM concentrations inhibited apoptosis 72 h post-treatment through caspase-7 inhibition. Although gene expression analysis through qRT-PCR demonstrates that risperidone did not affect caspase-7 gene expression.

Conclusions

This bioinformatics and experimental study suggests risperidone as a caspase-7 inhibitor with the potential to extend the lifespan of CHO cells and offers possible opportunities in biotechnology.

How to cite: Kafi S, Najafi S, Mahnam K, et al. Virtual screening and experimental analysis of caspase-7 inhibitors as candidates for extending the lifespan of CHO cells. Electron J Biotechnol 2024;71. https://doi.org/10.1016/j.ejbt.2024.04.007.

Background

To explore the efficacy of powder of ginsengporia and atractylodis macrocephalae combined with aerobic exercise in the treatment of obese T2DM and its influence on glucose and lipid metabolism indices.

Results

Both groups of patients were given metformin drug treatment. The control group was given aerobic exercise intervention, and the observation group was given ginsengporia and atractylodis macrocephalae on the basis of the control group. The total effective rate of the observation group (95.74%) was higher than that of the control group (82.98%) (P < 0.05). After treatment, the main symptoms of the observation group were less (P < 0.05), such as thirst and lack of desire to drink, fatigue, loss of appetite, limb numbness and tingling, sticky stool, scanty dark urine, and lower total score. FPG, 2hPG, and HbA1c were lower (P < 0.05), FINS, HOMA-IR, and HOMA-β were higher (P < 0.05), TG, TC, and LDL-C were lower, and HDL-C levels were higher in the observation group (P < 0.05). After treatment, Nesfatin-1 and adiponectin were lower, visfatin and leptin were higher (P < 0.05), and GIP and GLP-1 levels were higher in the observation group (P < 0.05). No significant difference was found in the incidence of adverse reactions between the two groups (P > 0.05).

Conclusions

Powder of ginsengporia and atractylodis macrocephalae combined with aerobic exercise is effective in treating obese T2DM, which can alleviate clinical symptoms and the disorder of glucose and lipid metabolism, and improve the islet function and gastrointestinal function.

How to cite: Liu G, Kanti Das S. Efficacy of powder of ginsengporia and atractylodis macrocephalae combined with aerobic exercise in the treatment of obese T2DM: A randomized controlled clinical trial. Electron J Biotechnol 2024;71. https://doi.org/10.1016/j.ejbt.2024.04.006.

Background

The Pacific white shrimp is one of the world’s most economically significant aquatic species, being one of the top three species cultured globally. However, the increasing incidence of diseases such as acute hepatopancreatic necrosis disease and hepatopancreatic microsporidia has led to a serious decline in shrimp production and severe economic losses. With the increasing demand for pathogen detection in shrimp farms, rapid DNA extraction technology has become more sophisticated. In this study, a rapid and crude method of extracting genomic DNA from shrimp muscle and hepatopancreas using Chelex-100 was established.

Results

DNA was successfully extracted from muscle and hepatopancreatic tissues using both the Chelex-100 method and commercial kits. The internal reference genes of shrimp were successfully amplified via PCR and real-time PCR using the obtained DNA samples. Moreover, a field assay was successfully conducted using real-time PCR and real-time enzymatic recombinase amplification (real-time ERA), indicating that the quality of the DNA extracted using Chelex-100 is sufficient for use in conjunction with nucleic acid amplification to detect pathogens in shrimps.

Conclusions

Chelex-100 is an efficient method for extracting DNA from shrimp muscle or hepatopancreas tissues, with a short extraction time, high extraction efficiency, and simple operation, making it appropriate for use in the detection of pathogens in shrimp.

How to cite: Yang H, Zhou Q, Hu J, et al. A Chelex-100-based rapid DNA extraction method and its application in the detection of shrimp pathogens. Electron J Biotechnol 2024;70. https://doi.org/10.1016/j.ejbt.2024.04.004.