Brazilian Journal of Microbiology最新文献

Chemical composition, antibacterial and antioxidant activities of Piper betle and Anethum graveolens essential oils against Methicillin-resistant Staphylococcus aureus clinical isolates.

IF 2.1 4区生物学

Brazilian Journal of Microbiology Pub Date : 2025-01-30 DOI: 10.1007/s42770-024-01567-9

Geetanjali Raikwar, Sumedha Mohan, Praveen Dahiya

{"title":"Chemical composition, antibacterial and antioxidant activities of Piper betle and Anethum graveolens essential oils against Methicillin-resistant Staphylococcus aureus clinical isolates.","authors":"Geetanjali Raikwar, Sumedha Mohan, Praveen Dahiya","doi":"10.1007/s42770-024-01567-9","DOIUrl":"https://doi.org/10.1007/s42770-024-01567-9","url":null,"abstract":"Methicillin-resistant Staphylococcus aureus (MRSA) is one of the leading causes of infection worldwide due to its antimicrobial resistance. Plant-derived essential oils (EOs) have undergone extensive observational and clinical research to explore their antimicrobial properties. The present study aimed to check mec A positive MRSA isolates using sequencing analysis, determination of chemical composition using gas chromatography-mass spectroscopy (GC-MS), antioxidant, and antimicrobial activity of Anethum graveolens and Piper betle EOs against the infectious agent MRSA. The result demonstrates a significant antibacterial activity of both essential oils against MRSA clinical isolates. GC-MS analysis of Piper betle showed (41.06%) 3-Allyl-6-methoxyphenol (Eugenol) as the major compound, whereas Anethum graveolens exhibited o-cymene (14.01%) abundantly. Piper betle essential oil retained appreciable levels of total phenolic (39.5 ± 10.9 mg/g of gallic acid equivalents) and flavonoid content (216 ± 145 mg quercetin equivalent/g), when compared to Anethum graveolens essential oil. A strong correlation was observed between antioxidant activity (DPPH, FRAP, and ABTS), total phenolic, and total flavonoid content in the Piper betle and displayed using principal component analysis (PCA) and a scatter matrix plot. Parallelly, clear morphological bacterial alterations were visualized by scanning electron microscopy after treating it with essential oils. MRSA showed malformed cell surfaces or broken cells with pore formation and septae. These findings imply that both essential oils are potential natural sources of antimicrobials against the MRSA superbug. They can also be used in combination therapies with other plant EOs or with traditional antibiotics to combat the rise of bacteria resistance.","PeriodicalId":9090,"journal":{"name":"Brazilian Journal of Microbiology","volume":" ","pages":""},"PeriodicalIF":2.1,"publicationDate":"2025-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143063646","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Biochemical features and biotechnological potential of a proteolytic extract from a psychrophilic Antarctic bacterium.

IF 2.1 4区生物学

Brazilian Journal of Microbiology Pub Date : 2025-01-29 DOI: 10.1007/s42770-024-01605-6

Franco Laureano, Susana Castro-Sowinski, Carolina Villadóniga

{"title":"Biochemical features and biotechnological potential of a proteolytic extract from a psychrophilic Antarctic bacterium.","authors":"Franco Laureano, Susana Castro-Sowinski, Carolina Villadóniga","doi":"10.1007/s42770-024-01605-6","DOIUrl":"https://doi.org/10.1007/s42770-024-01605-6","url":null,"abstract":"Proteases are hydrolases that act on peptide bonds, releasing amino acids and/or oligopeptides, and are involved in essential functions in all organisms. They represent an important segment of the global enzyme market, with applications in the food, leather, detergent, and pharmaceutical industries. Depending on their industrial use, proteases should exhibit high activity under extreme conditions, such as low temperatures, e.g. cold-active protease may have potential uses in the detergent industry. Cold-active enzymes show high catalytic constants (kcat) at low temperatures and thermolability, allowing their inactivation at moderate temperatures. This work aimed to characterize an extracellular proteolytic extract produced by an Antarctic isolate identified as Flavobacterium sp. strain AU13, and to evaluate its biotechnological potential as a detergent additive. By mass spectrometry analysis, we identified a major 50 kDa protease, with high identity with an epralysin from Pseudomonas fluorescens Pf0-1, an alkaline extracellular metalloprotease belonging to the serralysin subfamily. The AU13 proteolytic extract showed metalloprotease activity and, maximal activity over a wide pH range (pH 5 to 8); it also showed maximal activity at 40 °C, suggesting that this extracellular protease is a cold-active enzyme. The AU13 proteolytic extract demonstrated stable and compatible activity with surfactants and oxidants, making it a promising additive for commercial laundry detergents. Its ability to function effectively in cold-water washing conditions offers a significant advantage over conventional enzymes, potentially improving energy efficiency in industrial processes. The biochemical properties and performance of the AU13 proteolytic extract in the presence of laundry detergents, suggest that AU13 produces an extracellular protease with a biotechnological potential.","PeriodicalId":9090,"journal":{"name":"Brazilian Journal of Microbiology","volume":" ","pages":""},"PeriodicalIF":2.1,"publicationDate":"2025-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143057440","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Exploration of endophytic and rhizospheric bacteria of invasive plant Xanthium strumarium L. reveals their potential in plant growth promotion and bacterial wilt suppression.

IF 2.1 4区生物学

Brazilian Journal of Microbiology Pub Date : 2025-01-29 DOI: 10.1007/s42770-024-01599-1

Priyanka Das, Sampurna Kashyap, Indrani Sharma, Suvendra Kumar Ray, Niraj Agarwala

{"title":"Exploration of endophytic and rhizospheric bacteria of invasive plant Xanthium strumarium L. reveals their potential in plant growth promotion and bacterial wilt suppression.","authors":"Priyanka Das, Sampurna Kashyap, Indrani Sharma, Suvendra Kumar Ray, Niraj Agarwala","doi":"10.1007/s42770-024-01599-1","DOIUrl":"https://doi.org/10.1007/s42770-024-01599-1","url":null,"abstract":"Plant-associated microbiome plays important role in maintaining overall health of the host plant. Xanthium strumarium displaying resilience to various environmental fluctuations may harbor some bacterial isolates which can help this plant to grow worldwide. The present study aims to isolate endophytic and rhizospheric bacteria from X. strumarium and assess their plant growth-promoting and Ralstonia solanacearum antagonism activity. From a total of 148 isolated bacteria, 7 endophytic and 2 rhizospheric bacterial isolates were found to endow with significant in vitro plant growth promotion activities. The 16S rRNA gene sequence similarity of the 7 endophytic isolates has revealed these bacteria belonging to 5 genera viz. Curtobacterium, Pantoea, Pseudomonas, Microbacterium and Paracoccus whereas, the two rhizospheric isolates were identified as species of Ralstonia pickettii and Priestia megaterium. Maximum growth promotion was observed using the strains Pseudomonas fluorescens XSS6 and Microbacterium hydrothermale XSS20 in the assay conducted on tomato plants. In the in planta inhibition assay of R. solanacearum carried out in tomato seedlings using root bacterization method, Pseudomonas fluorescens XSS6 and Panotea vagans XSS3 showed antagonistic activity with biocontrol efficacy of 94.83% and 83.96%, respectively. GC-MS analysis detected several known antimicrobial compounds in the extract of the culture supernatant of Pseudomonas fluorescens XSS6 and Panotea vagans XSS3 strains, which may contribute to the inhibition of R. solanacearum by these strains. The results of our study indicated that the bacteria associated with X. strumarium exhibit multiple plant-beneficial effects. These bacteria have the potential to be developed as effective biofertilizers and biological control agents, promoting sustainable agriculture practices.","PeriodicalId":9090,"journal":{"name":"Brazilian Journal of Microbiology","volume":" ","pages":""},"PeriodicalIF":2.1,"publicationDate":"2025-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143057912","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Biogenic silver nanoparticle as an adjuvant in an S1 subunit recombinant vaccine.

IF 2.1 4区生物学

Brazilian Journal of Microbiology Pub Date : 2025-01-29 DOI: 10.1007/s42770-025-01613-0

Stella Buchhorn de Freitas, Amilton Clair Pinto Seixas Neto, Luciano Aparecido Panagio, Tiago Veiras Collares, Fabiana Kömmling Seixas, Fernanda Severo Sabedra Sousa, Daiane Drawanz Hartwig

{"title":"Biogenic silver nanoparticle as an adjuvant in an S1 subunit recombinant vaccine.","authors":"Stella Buchhorn de Freitas, Amilton Clair Pinto Seixas Neto, Luciano Aparecido Panagio, Tiago Veiras Collares, Fabiana Kömmling Seixas, Fernanda Severo Sabedra Sousa, Daiane Drawanz Hartwig","doi":"10.1007/s42770-025-01613-0","DOIUrl":"https://doi.org/10.1007/s42770-025-01613-0","url":null,"abstract":"Adjuvants are crucial for maintaining specific, protective, and long-lasting immunity. Here, we aimed to evaluate the antigenic and immunogenic activity of a recombinant form of the S1 domain of the Spike protein, associated with biogenic silver nanoparticles (bio-AgNP) and Alhydrogel® as an alternative and conventional adjuvant, respectively, for a SARS-CoV-2 subunit vaccine. We produced and evaluated the antigenicity of the recombinant S1 (rS1) protein by testing its recognition by antibodies present in SARS-CoV-2 positive human serum. The immunogenicity of the rS1 protein was assessed in vivo by its ability to induce antibody production in mice. Furthermore, we sought to establish the types of humoral and cellular immune responses generated through intramuscular immunization in BALB/c mice with the rS1 + bio-AgNP vaccine. The recombinant S1 (rS1) protein was successfully cloned, expressed in Escherichia coli, and confirmed to have strong antigenic potential, being recognized by human antibodies up to a 1:51,200 serum dilution. In vivo assays showed that vaccines using rS1 with either bio-AgNP or Alhydrogel® as adjuvants induced high and consistent antibody titers (1:51,200) and a range of antibody isotypes in mice. Cellular immune response analysis revealed significant IL-10 expression with rS1 + bio-AgNP and both IL-10 and TNFα expression with rS1 + Alhydrogel®. These results support rS1 + bio-AgNP as a promising vaccine candidate for future development, highlighting its potential not only as a viable alternative to traditional adjuvants but also as an innovative contribution to advancing more effective and accessible immunological strategies.","PeriodicalId":9090,"journal":{"name":"Brazilian Journal of Microbiology","volume":" ","pages":""},"PeriodicalIF":2.1,"publicationDate":"2025-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143057977","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

An assessment of plant growth and physiological responses in annual crops grown in P deficient soils inoculated with indigenous arbuscular mycorrhizal fungi.

IF 2.1 4区生物学

Brazilian Journal of Microbiology Pub Date : 2025-01-29 DOI: 10.1007/s42770-025-01618-9

Tancredo Souza, Josefa Bruna Lima Dos Santos, Diego Silva Batista

{"title":"An assessment of plant growth and physiological responses in annual crops grown in P deficient soils inoculated with indigenous arbuscular mycorrhizal fungi.","authors":"Tancredo Souza, Josefa Bruna Lima Dos Santos, Diego Silva Batista","doi":"10.1007/s42770-025-01618-9","DOIUrl":"https://doi.org/10.1007/s42770-025-01618-9","url":null,"abstract":"The influence of arbuscular mycorrhizal fungi (AMF) inoculation on the growth and physiology of Phaseolus vulgaris L. and Zea mays L. in the Brazilian tropical seasonal dry forest is not well known. We examined the effects of indigenous AMF species inoculation on these two annual crops at phosphorus-deficient soils under glasshouse conditions. The AMF species used as inoculum were collected from the root zone of Mimosa tenuiflora, a native plant of the Caatinga ecoregion. The inoculum was propagated in plastic pots with Trifolium sp. as the host plant. We investigated the effect of four treatments: (1) Control- non-inoculated; (2) Acaulospora inoculum- inoculation with A. tuberculata; (3) Gigaspora inoculum- inoculation with G. gigantea; and (4) Mixed inoculum- inoculation with both Acaulospora and Gigaspora species. We found that inoculation with the indigenous AMF community enhanced plant dry biomass, plant P concentration, root colonization, and physiological responses for both model plants. This study highlighted the importance of exploring the P molecule as an integral element in plant development and presents the AMF inoculation as an efficient alternative to overcome low-P conditions, leading to improved nutrient acquisition in low-fertility soils, better growth conditions through enhanced physiological responses, and ultimately increased plant dry biomass.","PeriodicalId":9090,"journal":{"name":"Brazilian Journal of Microbiology","volume":" ","pages":""},"PeriodicalIF":2.1,"publicationDate":"2025-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143058080","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Detection and characterization of pathogenic Bacillus haynesii from Tribulus terrestris extract: ways to reduce its levels.

IF 2.1 4区生物学

Brazilian Journal of Microbiology Pub Date : 2025-01-29 DOI: 10.1007/s42770-024-01608-3

Priti Darne, Shital Palghadmal, Nisha Mehta, Sriram Padmanabhan

{"title":"Detection and characterization of pathogenic Bacillus haynesii from Tribulus terrestris extract: ways to reduce its levels.","authors":"Priti Darne, Shital Palghadmal, Nisha Mehta, Sriram Padmanabhan","doi":"10.1007/s42770-024-01608-3","DOIUrl":"https://doi.org/10.1007/s42770-024-01608-3","url":null,"abstract":"Plant parts such as roots, bark, leaves, flowers, and fruits that hold ethnopharmacological significance are naturally prone to microbial contamination, influenced by environmental factors like moisture and humidity. This study focuses on assessing the microbial load in the raw material of Tribulus terrestris (TT). The primary bacterium isolated from the pulverized raw material was identified as Bacillus haynesii through 16S rRNA sequencing. Biochemical assays revealed the organism's ability to utilize lysine and ornithine, produce urease, and generate hydrogen sulfide. The bacterium exhibited resistance to multiple antibiotics and caused 21.5% hemolysis in RBC lysis assays. To reduce microbial contamination, Glutaraldehyde (GA) and polyhexamethylene biguanide (PHMB) were tested, with GA at 1% reducing the microbial load by 99% without affecting the yield (0.5%) or bioactive saponin content. High-Performance Liquid Chromatography (HPLC) confirmed the absence of residual GA, ensuring an eco-friendly and safe process. This highlights the importance of quality control measures, including Hazard Analysis and Critical Control Points (HACCP) regulations, in maintaining the integrity of herbal extracts.","PeriodicalId":9090,"journal":{"name":"Brazilian Journal of Microbiology","volume":" ","pages":""},"PeriodicalIF":2.1,"publicationDate":"2025-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143057909","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Biofilm formation by the plant growth promoting bacterium Bacillus cereus (EB-40).

IF 2.1 4区生物学

Brazilian Journal of Microbiology Pub Date : 2025-01-29 DOI: 10.1007/s42770-025-01617-w

Débora Francine Gomes Silva Pereira, Silvia Nietsche, Adelica Aparecida Xavier, Maria Olívia Mercadante-Simões, Marlon Cristian Toledo Pereira

引用次数: 0

Bioactivities of secondary metabolites of two actinomycetes Streptomyces parvulus GloL3, and Streptomyces lienomycini SK5, endophytes of two Indian medicinal herbs- Globba marantina L. and Selaginella kraussiana (Kunze) A. Braun.

IF 2.1 4区生物学

Brazilian Journal of Microbiology Pub Date : 2025-01-29 DOI: 10.1007/s42770-025-01615-y

Hiran Kanti Santra, Debdulal Banerjee

{"title":"Bioactivities of secondary metabolites of two actinomycetes Streptomyces parvulus GloL3, and Streptomyces lienomycini SK5, endophytes of two Indian medicinal herbs- Globba marantina L. and Selaginella kraussiana (Kunze) A. Braun.","authors":"Hiran Kanti Santra, Debdulal Banerjee","doi":"10.1007/s42770-025-01615-y","DOIUrl":"https://doi.org/10.1007/s42770-025-01615-y","url":null,"abstract":"Endophytic actinomycetes are potential sources of novel pharmaceutically active metabolites, significantly advancing natural product research. In the present investigation, secondary metabolites from two endophytic actinomycetes, Streptomyces parvulus GloL3, and Streptomyces lienomycini SK5, isolated from medicinal plant taxa, Globba marantina, and Selaginella kraussiana, exhibited broad-spectrum bioactivity. Ethyl Acetate (EA) extract of SK5 showed antimicrobial activity against nine human pathogens, including Methicillin-resistant Staphylococcus aureus (MRSA), Candida tropicalis, and C. albicans, with a minimum microbicidal concentration (MMC) of 50-300 µg mL-1. It healed the MRSA-mediated wounds in Swiss albino mice in vivo. EA extracts dissociate the pathogenic cell membranes and cause leakage of biomacromolecules-nucleic acid, protein, and potassium ions. Also, critical housekeeping enzymes involved in the cellular respiratory mechanisms of the pathogens were blocked. GloL3 has antioxidative potentialities against DPPH, ABTS, FRAP, and H2O2 free radical generators with an IC50 value of 21.18 ± 0.33, 43.58 ± 0.91, 88.24 ± 1.24, and 111.03 ± 6.42 µg mL-1. It improves the enzymatic antioxidant parameters in treated peritoneal macrophage cells of Swiss albino mice. Constituents of the EA extracts of GloL3 and SK5 are bactobolin, actinobolin, 5-(2-aminoethyl)-1 H imidazole-2-carbaldehyde, isovaleric acid, fulvic acid, phenol, 4-[2-(methylamino) ethyl]-, eicosanoic acid, heptadecanoic acid, etc. The present findings suggest that metabolites from the endophytes of medicinal plants hold potent pharmaceutical utilities.","PeriodicalId":9090,"journal":{"name":"Brazilian Journal of Microbiology","volume":" ","pages":""},"PeriodicalIF":2.1,"publicationDate":"2025-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143058081","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Soil and climatic conditions determine the rhizobia in association with Phaseolus vulgaris in southern Brazil.

IF 2.1 4区生物学

Brazilian Journal of Microbiology Pub Date : 2025-01-23 DOI: 10.1007/s42770-025-01621-0

Tales Gustavo Ferreto de Araújo, Elisete Pains Rodrigues, Mariangela Hungria, Fernando Gomes Barcellos

{"title":"Soil and climatic conditions determine the rhizobia in association with Phaseolus vulgaris in southern Brazil.","authors":"Tales Gustavo Ferreto de Araújo, Elisete Pains Rodrigues, Mariangela Hungria, Fernando Gomes Barcellos","doi":"10.1007/s42770-025-01621-0","DOIUrl":"https://doi.org/10.1007/s42770-025-01621-0","url":null,"abstract":"The common bean (Phaseolus vulgaris L.) plays a significant economic and social role in Brazil. However, the national average yield remains relatively low, largely because most bean cultivation is undertaken by small-scale farmers. In this context, biological nitrogen fixation (BNF) is an effective strategy for improving crop yield. Therefore, it is important to identify novel rhizobial strains well suited to local climatic conditions. This study used Phaseolus vulgaris as a trap plant in soils from three distinct conservation areas (Ponta Grossa, Ortigueira, and Londrina) within Paraná State, Brazil. The soil chemical analysis revealed that the pH values in the Ponta Grossa and Ortigueira regions were low, whereas the Ortigueira region exhibited elevated aluminum levels. A total of 94 strains were obtained from the nodules of plants and subjected to analysis for their morphological and genetic properties. No nodules were observed in the Ortigueira region. In the Ponta Grossa region, most of the strains were identified as belonging to the genus Paraburkholderia, whereas all strains from Londrina were identified as Rhizobium. The 16S rRNA gene phylogenetic analysis revealed a high degree of genetic similarity between the Paraburkholderia and Rhizobium strains. These findings indicate that soil chemical properties (pH and aluminum level) and climate conditions may have a significant impact on the symbiotic association between rhizobia and common bean.","PeriodicalId":9090,"journal":{"name":"Brazilian Journal of Microbiology","volume":" ","pages":""},"PeriodicalIF":2.1,"publicationDate":"2025-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143022094","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Optimizing proinsulin production in E. coli BL21 (DE3) using taguchi method and efficient one-step insulin purification by on-column enzymatic cleavage.

IF 2.1 4区生物学

Brazilian Journal of Microbiology Pub Date : 2025-01-23 DOI: 10.1007/s42770-025-01614-z

Shahin Hadadian, Mina Sepahi, Samin Sedighi

{"title":"Optimizing proinsulin production in E. coli BL21 (DE3) using taguchi method and efficient one-step insulin purification by on-column enzymatic cleavage.","authors":"Shahin Hadadian, Mina Sepahi, Samin Sedighi","doi":"10.1007/s42770-025-01614-z","DOIUrl":"https://doi.org/10.1007/s42770-025-01614-z","url":null,"abstract":"Diabetes is a critical worldwide health problem. Numerous studies have focused on producing recombinant human insulin to address this issue. In this research, the process factors of production of recombinant His-tagged proinsulin in E. coli BL21 (DE3) strain were studied. Bacterial culture factors with significant effects on the amount of produced recombinant proinsulin were screened using a Taguchi L8 orthogonal array. Proinsulin expression was conducted under predicted optimal conditions. The folded impure His-tagged proinsulin was purified using immobilized metal ion affinity chromatography (IMAC). A novel IMAC sequence order combined with the use of non-His-tagged C-peptide cleavage enzymes followed by His- tagged enterokinase enzyme enabled simultaneous protein purification and elimination of C-peptide and His-tag in just one step. Statistical analysis revealed that the amount of produced proinsulin was significantly affected by several factors including the post-induction incubation temperature, Isopropyl ß-D-1-thiogalactopyranoside (IPTG) concentration, pre-induction incubation temperature, the glucose concentration, bacterial cell population at induction step, and the time of harvesting. The optimized model resulted in an empirical maximum proinsulin concentration of 254.5 ± 11.7 µg/ml. The high purity of the purified insulin (> 96% by SDS-PAGE) indicated that applied IMAC sequence order could be considered an efficient technique for on-column cleavage and insulin purification.","PeriodicalId":9090,"journal":{"name":"Brazilian Journal of Microbiology","volume":" ","pages":""},"PeriodicalIF":2.1,"publicationDate":"2025-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143022092","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0