Electronic Journal of Biotechnology最新文献

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

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

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

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

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

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.

Background

Cold damage of maize during germination is a global problem; it occurs frequently in northeast China, and leads to a large-scale reduction in yield. Low temperature tolerance of maize in germination is a complex quantitative trait controlled by multigenes, and no major QTLs or key genes have been identified.

Results

An F₂ isolation population with S319 and R144 as parents was constructed. The bulked segregant analysis (BSA) and specific-locus amplified fragment-sequencing (SLAF-seq) methods were applied to locate the chromosomal association regions related to low-temperature tolerance of maize during germination. Sequencing obtained 221.72 Gbp clean data, with an average sequencing depth of 25.96X. Four candidate regions associated with low-temperature tolerance trait of maize in germination were obtained, with a total length of 25.71 Mb and 1513 annotated genes, including 456 nonsynonymous mutant genes and 111 frameshift mutant genes.

Conclusions

This study aimed to lay the foundation for the mining of candidate genes of low-temperature tolerance in maize during germination, and accelerate the process of targeted improvement of maize low-temperature tolerance molecular marker-assisted breeding.

How to cite: Yu T, Zhang J, Cao J, et al. QTL analysis of low temperature tolerance in maize germination by SLAF-seq and BSA technique. Electron J Biotechnol 2024;70. https://doi.org/10.1016/j.ejbt.2024.04.003.

Background

Cancer continues to be one of the greatest challenges in modern medicine and is second only to cardiovascular disease as the main cause of death. Breast cancer in particular is responsible for 15% of deaths in women. In this study, Lactobacillus bulgaricus was microencapsulated using a chitosan/alginate mixture. Parameters such as chitosan, alginate, and L. bulgaricus populations were optimized using Design Expert software. The responses were loading efficiency, particle size, release, and ζ-potential. Subsequently, the cytotoxicity of the optimized ratio of chitosan/alginate nanoparticles was investigated on MCF-7 cancer cells.

Results

The research revealed that optimal conditions for the mentioned variables were a chitosan concentration of 1% w/w, an alginate concentration of 1% w/w, and a L. bulgaricus count of 8.15 CFU/ml. Following numerical optimization, the loading efficacy = 91.15%, the release = 71.55%, the polydispersity index = 0.11, and the ζ-potential = 61.94 based on numerical optimization. Findings revealed that miracle drinks with L. bulgaricus-loaded chitosan/alginate microcapsule ratios exhibited toxic and potential apoptotic effects on MCF-7 cancer cells. This study showed that a miracle drink prepared with the optimal ratio of probiotic nanoparticles stops cells in the S and G2/M phases.

Conclusions

The results show that Miracle drink supplemented with L. bulgaricus loaded-chitosan/alginate nanoparticles has a toxic and lethal effect on MCF-7 cancer cells. This compound can be suggested and used as an alternative candidate or complementary cancer therapy.

How to cite: Jovaini K, Mortazavian Farsani SAM, Aghaee-Bakhtiari SH, et al. Miracle drink supplemented with L.bulgaricus loaded-chitosan/alginate nanoparticles as a medicinal food for control of MCF7 cancer cells. Electron J Biotechnol 2024;69. https://doi.org/10.1016/j.ejbt.2024.04.002.

Background

The lack of reference genes makes it difficult to conduct molecular biology research on the plant genus, Clematis L. Clematis lanuginosa belongs to Sect. Viticella DC of Clematis L. It is also an important ornamental cultivated variety parent of the early and late large-flowered groups. Studying the reference genes of C. lanuginosa in different tissues will provide a theoretical basis for the reference selection of early and late large-flowered groups of Clematis, which could promote research progress on molecular biology of ornamental Clematis.

Results

The roots, stems, leaves, sepals, stamens, and carpels of C. lanuginosa were used as research materials, and seven candidate reference genes were used for quantitative real-time PCR analysis. Comprehensive stability analysis using geNorm, NormFinder, BestKeeper, and RefFinder software showed that suitable reference genes in C. lanuginosa root, stem, and leaf were PP2A-2 and UBC34; and in floral tissue were UBC34, PP2A-2, and ARP7. These reference genes can be used as internal reference either alone or in combination. The pairwise variation value evaluated with geNorm software showed that two internal reference genes were needed for gene expression correction in the tissues. In the floral organs, three reference genes were required for gene expression correction.

Conclusions

Our results provide a foundation for future gene expression analysis of C. lanuginosa and guidance for the screening of reference genes in Clematis.

How to cite: Li Q, Wang S, Lv F, et al. Selection and validation of reference genes for quantitative real-time PCR in different tissues of Clematis lanuginosa. Electron J Biotechnol 2024;70. https://doi.org/10.1016/j.ejbt.2024.04.005.