Journal, genetic engineering & biotechnology最新文献

{"title":"Intrinsic disorder in the open reading frame 2 of hepatitis E virus: a protein with multiple functions beyond viral capsid.","authors":"Zoya Shafat,&nbsp;Anwar Ahmed,&nbsp;Mohammad K Parvez,&nbsp;Shama Parveen","doi":"10.1186/s43141-023-00477-x","DOIUrl":"https://doi.org/10.1186/s43141-023-00477-x","url":null,"abstract":"<p><strong>Background: </strong>Hepatitis E virus (HEV) is the cause of a liver disease hepatitis E. The translation product of HEV ORF2 has recently been demonstrated as a protein involved in multiple functions besides performing its major role of a viral capsid. As intrinsically disordered regions (IDRs) are linked to various essential roles in the virus's life cycle, we analyzed the disorder pattern distribution of the retrieved ORF2 protein sequences by employing different online predictors. Our findings might provide some clues on the disorder-based functions of ORF2 protein that possibly help us in understanding its behavior other than as a HEV capsid protein.</p><p><strong>Results: </strong>The modeled three dimensional (3D) structures of ORF2 showed the predominance of random coils or unstructured regions in addition to major secondary structure components (alpha helix and beta strand). After initial scrutinization, the predictors VLXT and VSL2 predicted ORF2 as a highly disordered protein while the predictors VL3 and DISOPRED3 predicted ORF2 as a moderately disordered protein, thus categorizing HEV-ORF2 into IDP (intrinsically disordered protein) or IDPR (intrinsically disordered protein region) respectively. Thus, our initial predicted disorderness in ORF2 protein 3D structures was in excellent agreement with their predicted disorder distribution patterns (evaluated through different predictors). The abundance of MoRFs (disorder-based protein binding sites) in ORF2 was observed that signified their interaction with binding partners which might further assist in viral infection. As IDPs/IDPRs are targets of regulation, we carried out the phosphorylation analysis to reveal the presence of post-translationally modified sites. Prevalence of several disordered-based phosphorylation sites further signified the involvement of ORF2 in diverse and significant biological processes. Furthermore, ORF2 structure-associated functions revealed its involvement in several crucial functions and biological processes like binding and catalytic activities.</p><p><strong>Conclusions: </strong>The results predicted ORF2 as a protein with multiple functions besides its role as a capsid protein. Moreover, the occurrence of IDPR/IDP in ORF2 protein suggests that its disordered region might serve as novel drug targets via functioning as potential interacting domains. Our data collectively might provide significant implication in HEV vaccine search as disorderness in viral proteins is related to mechanisms involved in immune evasion.</p>","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":"21 1","pages":"33"},"PeriodicalIF":0.0,"publicationDate":"2023-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10018590/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9139723","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Xylanopectinolytic enzymes by marine actinomycetes from sediments of Sarena Kecil, North Sulawesi: high potential to produce galacturonic acid and xylooligosaccharides from raw biomass.","authors":"Hana Nadhifah,&nbsp;Nanik Rahmani,&nbsp;Wibowo Mangunwardoyo,&nbsp;Yopi,&nbsp;Akhirta Atikana,&nbsp;Shanti Ratnakomala,&nbsp;Puspita Lisdiyanti","doi":"10.1186/s43141-023-00488-8","DOIUrl":"https://doi.org/10.1186/s43141-023-00488-8","url":null,"abstract":"<p><strong>Background: </strong>Actinomycetes isolated from marine habitats are known to have the potential for novel enzymes that are beneficial in the industry. In-depth knowledge is necessary given the variety of this bacterial group in Indonesia and the lack of published research. Actinomycetes isolates (BLH 5-14) obtained from marine sediments of Sarena Kecil, Bitung, North Sulawesi, Indonesia, showed an ability to produce pectinase and xylanase that have equal or even higher potential for pectic-oligosaccharides (POS) and xylooligosaccharides (XOS) production from raw biomass than from commercial substrates. This study's objective was to characterize both enzymes to learn more for future research and development.</p><p><strong>Results: </strong>Pectinase had the highest activity on the 6^th day (1.44±0.08 U/mL) at the optimum pH of 8.0 and optimum temperature of 50 °C. Xylanase had the maximum activity on the 6^th day (4.33±0.03 U/mL) at optimum pH 6.0 and optimum temperature 60 °C. Hydrolysis and thin layer chromatography also showed that pectinase was able to produce monosaccharides such as galacturonic acid (P1), and xylanase was able to yield oligosaccharides such as xylotriose (X3), xylotetraose (X4), and xylopentaose (X5). BLH 5-14 identified as the genus Streptomyces based on the 16S rDNA sequences and the closely related species Streptomyces tendae (99,78%).</p><p><strong>Conclusions: </strong>In the eco-friendly paper bleaching industry, Streptomyces tendae has demonstrated the potential to create enzymes with properties that can be active in a wide range of pH levels. The oligosaccharides have the potential as prebiotics or dietary supplements with anti-cancer properties. Further research is needed to optimize the production, purification, and development of the application of pectinase and xylanase enzymes produced by Actinomycetes isolates.</p>","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":"21 1","pages":"31"},"PeriodicalIF":0.0,"publicationDate":"2023-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10017887/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9131711","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An assessment for in vitro propagation and genetic stability of Phoebe goalparensis Hutchinson, an endemic valuable timber tree of North East India.","authors":"Kalpataru Dutta Mudoi,&nbsp;Barbi Gogoi,&nbsp;Gitasree Borah,&nbsp;Marine Hussain,&nbsp;Tabassum Tasfia,&nbsp;Krishnakhi Borah,&nbsp;Himangshu Lekhak,&nbsp;Siddhartha Proteem Saikia","doi":"10.1186/s43141-023-00487-9","DOIUrl":"https://doi.org/10.1186/s43141-023-00487-9","url":null,"abstract":"<p><strong>Background: </strong>Phoebe goalparensis is an endemic forest species of North East India that belongs to Lauraceae family. P. goalparensis is used as timbers yielding plants for commercial importance in the local furniture markets of North East India. A rapid in vitro micropropagation protocol was established by using apical and axillary shoot tips on Murashige and Skoog medium with varied concentrations of plant growth regulators.</p><p><strong>Results: </strong>In this study, 5.0 mg/l BAP augmented medium was chosen as the best for shoot multiplication of the plant. However, IBA (2.0 mg/l) was the most responsive for root induction. Moreover, 70% of root induction was recorded during rooting experiment and 80-85% survivability was observed during the acclimatization of this species. Clonal fidelity of P. goalparensis was determined with ISSR marker and it was observed that in vitro raised plantlets were polymonomorphic.</p><p><strong>Conclusion: </strong>Hence, an efficient protocol with high proliferation and rooting was established for P. Goalparensis that could aid in massive propagation in future.</p>","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":"21 1","pages":"29"},"PeriodicalIF":0.0,"publicationDate":"2023-03-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10006360/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9099664","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Insight study on synthesis and antibacterial mechanism of silver nanoparticles prepared from indigenous plant source of Jharkhand.","authors":"Koel Mukherjee,&nbsp;Namrata Bhagat,&nbsp;Madhubala Kumari,&nbsp;Arnab Roy Choudhury,&nbsp;Biplab Sarkar,&nbsp;Barnali Dasgupta Ghosh","doi":"10.1186/s43141-023-00463-3","DOIUrl":"https://doi.org/10.1186/s43141-023-00463-3","url":null,"abstract":"<p><strong>Background: </strong>The Ag-NPs by green synthesis has a notable interest because of their eco-friendliness, economic views, feasibility, and applications in a wide range. Herein, native plants of Jharkhand (Polygonum plebeium, Litsea glutinosa, and Vangueria spinosus) were selected for the current work of Ag-NP synthesis and further antibacterial activity. Green synthesis was performed for Ag-NPs using Silver nitrate solution as precursor and the dried leaf extract performs as a reductant and stabilizer here.</p><p><strong>Result: </strong>Visually Ag-NP formation was observed along with a colour change and confirmed by UV-visible spectrophotometry on which an absorbance peak occurs at around 400-450nm. Further characterization was done on DLS, FTIR, FESEM, and XRD. Size around 45-86 nm of synthesized Ag-NPs was predicted through DLS. The synthesized Ag-NPs exhibited significant antibacterial activity against Bacillus subtilis (Gram-positive bacteria) and Salmonella typhi (Gram-negative bacteria). The finest antibacterial activity was disclosed by the Ag-NPs synthesized by Polygonum plebeium extract. The diameter of the zone of inhibition in the bacterial plate measured was 0-1.8 mm in Bacillus and 0-2.2 mm in Salmonella typhi. Protein-Protein interaction study was performed to study the effect of Ag-NPs towards different antioxidant enzyme system of bacterial cell.</p><p><strong>Conclusion: </strong>Present work suggest the Ag-NPs synthesized from P. plebeium were more stable for long term and might have prolonged antibacterial activity. In the future, these Ag-NPs can be applied in various fields like antimicrobial research, wound healing, drug delivery, bio-sensing, tumour/cancer cell treatment, and detector (detect solar energy). Schematic representation of Ag-NPs green synthesis, characterization, antibacterial activity and at the end, in silico study to analyse the mechanism of antibacterial activity.</p>","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":"21 1","pages":"30"},"PeriodicalIF":0.0,"publicationDate":"2023-03-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10006383/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9105162","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Purification, characterization, and enzyme kinetics of a glutathione S transferase from larvae of the camel tick Hyalomma dromedarii.","authors":"Hassan M M Masoud, Mohamed S Helmy, Doaa A Darwish, Mahmoud A Ibrahim","doi":"10.1186/s43141-023-00486-w","DOIUrl":"10.1186/s43141-023-00486-w","url":null,"abstract":"<p><strong>Background: </strong>Glutathione s-transferases (GSTs) perform an essential role in detoxification of xenobiotics and endogenous compounds via their conjugation to reduce glutathione.</p><p><strong>Results: </strong>A GST enzyme, designated tick larvae glutathione S transferase (TLGST), was purified from larvae of the camel tick Hyalomma dromedarii via ammonium sulfate precipitation, glutathione-Sepharose affinity column and Sephacryl S-300 chromatography. TLGST-specific activity was found to be 1.56 Umg^-1 which represents 39 folds and 32.2% recovery. The molecular weight of TLGST purified from camel tick larvae was found as 42 kDa by gel filtration. TLGST has a pI value of 6.9 and was found a heterodimeric protein of 28 and 14 kDa subunits as detected on SDS-PAGE. The Lineweaver-Burk plot calculated the km for CDNB to be 0.43 mM with Vmax value of 9.2 Umg^-1. TLGST exhibited its optimal activity at pH 7.9. Co²⁺, Ni²⁺ and Mn²⁺ increased the activity of TLGST while Ca²⁺, Cu²⁺, Fe²⁺ and Zn²⁺ inhibited it. TLGST was inhibited by cumene hydroperoxide, p-hydroxymercuribenzoate, lithocholic acid, hematin, triphenyltin chloride, p-chloromercuribenzoic acid (pCMB), N-p-Tosyl-L-phenylalanine chloromethyl ketone (TPCK), iodoacetamide, EDTA and quercetin. pCMB inhibited TLGST competitively with Ki value of 0.3 mM.</p><p><strong>Conclusions: </strong>These findings will help to understand the various physiologic conditions of ticks and targeting TLGST could be significant tool for development of prospective vaccines against ticks as a bio-control strategy to overcome the rapid grows in pesticide-resistant tick populations.</p>","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":"21 1","pages":"28"},"PeriodicalIF":3.6,"publicationDate":"2023-03-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9995618/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9139762","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Extract of Bletilla formosana callus elevates cellular antioxidative activity via Nrf2/HO-1 signaling pathway and inhibits melanogenesis in zebrafish.","authors":"Ruei-Ting Wang,&nbsp;Jui-Hung Yen,&nbsp;Yi-Chi Liao,&nbsp;Yi-Zhen Li,&nbsp;Wei-Ping Wang","doi":"10.1186/s43141-023-00482-0","DOIUrl":"https://doi.org/10.1186/s43141-023-00482-0","url":null,"abstract":"<p><strong>Background: </strong>Bletilla species are endangered terrestrial orchids used in natural skin care formulas in Asia for a long history. In order to explore the bioactivity potential of Bletilla species as a cosmetic ingredient in a sustainable resource manner, the callus of Bletilla formosana (Hayata) Schltr. was established and extracted by an eco-friendly supercritical fluid CO₂ extraction (SFE-CO₂) method. The intracellular reactive oxygen species (ROS) scavenging activity and antioxidation-related gene expression of the callus extract were evaluated in both Hs68 fibroblast cells and HaCaT keratinocytes. The melanogenesis-inhibitory effect was investigated in B16F10 melanoma cells and in an in vivo zebrafish model.</p><p><strong>Results: </strong>The calli of B. formosana were propagated for 10-15 generations with a consistent yellow friable appearance and then subjected to SFE-CO₂ extraction to obtain a yellow pasty extract. Obvious intracellular ROS scavenging activity of the extract was detected in both Hs68 and HaCaT cells with 64.30 ± 8.27% and 32.50 ± 4.05% reduction at the concentration of 250 μg/mL. Moreover, marked expression levels of heme oxygenase-1 (HO-1) and (NAD(P)H) quinone oxidoreductase-1 (NQO1) genes were detected after 6-h and 24-h treatments. These results indicate the cellular antioxidative activity of B. formosana callus extract was probably activated via the nuclear factor erythroid 2-related factor 2 (Nrf2)/HO-1 signaling pathway. Melanogenesis-inhibitory effect of the extract was observed in α-MSH stimuli-inducing B16F10 cells with 28.46% inhibition of intracellular melanin content at the concentration of 50 μg/ml. The effect was confirmed with in vivo zebrafish embryos that showed a relative pigmentation density of 80.27 ± 7.98% at the concentration of 100 μg/mL without toxicity.</p><p><strong>Conclusion: </strong>Our results shed light on a sustainable utilization of Bletilla species as a potential ingredient for skin.</p>","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":"21 1","pages":"26"},"PeriodicalIF":0.0,"publicationDate":"2023-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9989080/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9131358","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Gold nanoparticles combined baker's yeast as a successful approach for breast cancer treatment.","authors":"Amany Elwakkad,&nbsp;Amina A Gamal El Din,&nbsp;Hisham A Saleh,&nbsp;Noha E Ibrahim,&nbsp;Mohamed A Hebishy,&nbsp;Hagar H Mourad,&nbsp;Mahitab I El-Kassaby,&nbsp;Howida Sayed Abou-Seif,&nbsp;Ghada M Elqattan","doi":"10.1186/s43141-023-00481-1","DOIUrl":"https://doi.org/10.1186/s43141-023-00481-1","url":null,"abstract":"<p><strong>Background: </strong>Saccharomyces cerevisiae (S. cerevisiae) has been demonstrated in vitro to sensitize several breast cancer cell lines and to be a safe, non-toxic drug with anti-skin cancer action in mice. Furthermore, plasmonic photothermal treatment using gold nanorods has been authorized as a novel method for in vitro and in vivo cancer therapy.</p><p><strong>Results: </strong>When compared to tumor-free rats, the treatment with S. cerevisiae conjugated to gold nanospheres (GNSs) lowered Bcl-2 levels while increasing FasL, Bax, cytochrome c, and caspases 8, 9, and 3 levels. Histopathological results showed changes reflecting the ability of nanogold conjugated heat-killed yeast to induce apoptosis is greater than heat-killed yeast alone as the nanogold conjugated with heat-killed yeast showed no tumor, no hyperplasia, no granulation tissue formation, no ulceration, and no suppuration. Nanogold conjugated with heat-killed yeast-treated breast cancer group displayed normal levels of ALT and AST, indicating relatively healthy hepatic cells.</p><p><strong>Conclusion: </strong>Our results proved that nanogold conjugated heat-killed yeast can initiate apoptosis and can be used as a safe non-invasive method for breast cancer treatment more effectively than the yeast alone. This, in turn, gives us new insight and a future hope for the first time that breast cancer can be treated by non-invasive, simple, safe, and naturally originated method and achieves a hopeful treatment and a novel method for in vivo cancer therapy.</p>","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":"21 1","pages":"27"},"PeriodicalIF":0.0,"publicationDate":"2023-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9989084/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9131357","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cultivation of Pseudochlorella pringsheimii for biodiesel production in a scalable indoor photobioreactor: case studies from Egypt.","authors":"Hanaa Abd El Baky,&nbsp;Gamal El Baroty","doi":"10.1186/s43141-022-00450-0","DOIUrl":"https://doi.org/10.1186/s43141-022-00450-0","url":null,"abstract":"<p><strong>Background: </strong>Enhancement of lipid accumulation is the major strategy to improve the commercial feasibility of microalgae as a source for biodiesel production. Pseudochlorella pringsheimii (Formally was named as Chlorella ellipsoidea) green microalgae strain was chosen with respect to their ability as a potential source to produce high lipids content, could be used for the production of biofuel, which can be an alternative renewable energy source instead of fossil fuels.</p><p><strong>Results: </strong>Initially, the Pseudochlorella pringsheimii microalgae was evaluated on the basis of tested at Lab scales 2 L by applicable different nutrient individual of N, P, Fe conditions in BBM medium concentrations for choosing the best concentrations induce lipid contents and productivity to cultivate in large scale in the 2000 L PBR. The suitable concentrations of nutrients with highest lipid contents were obtained under deficient of nitrogen (1.25 gL^-1, limited N) and phosphorus (0.1 mg L^-1, limited P) coupled with high iron concentration (10 mg L, rich Fe) and CO₂ (6%). Therefore, their collective of nutrients was applied to culture of microalgae cells at large scale in 2000 L photobioreactor (PBR model), which, this techniques was used to quantify high lipid contents (25% w/w) and high lipid productivity (74.07 mgL^-1 day^-1). The inducted lipid conversion to biodiesel via transestrification process was 91.54 ± 1.43%. The fatty acid methyl esters (FAMEs profile by means of GC/MS resulted in C16:0, C18:1, C18:2, C18:3 as a main constituents. With regard to physical-chemical property (such as density, kinematic viscosity, gravity, and certain number), the Pseudochlorella pringsheimii biodiesel have biofuel properties, in accordance with appropriate biodiesel properties, as ASTM and EU standards, that thereby referring to high quality biodiesel.</p><p><strong>Conclusions: </strong>Pseudochlorella pringsheimii cultured in large scale in photobioreactor under stress condition have a high potential of lipids production with high quality of FAMEs that can be used as a promising biodiesel fuel. It has also a potential to be applied for commercialization based on the techno-economic and environmental impacts.</p>","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":"21 1","pages":"25"},"PeriodicalIF":0.0,"publicationDate":"2023-03-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9981844/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10820720","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Selective isolation and genomic characterization of biopolymer producer-a novel feature of halophile Brachybacterium paraconglomeratum MTCC 13074.","authors":"Teja Mandragutti,&nbsp;G Sudhakar","doi":"10.1186/s43141-023-00484-y","DOIUrl":"https://doi.org/10.1186/s43141-023-00484-y","url":null,"abstract":"<p><strong>Background: </strong>Biopolymers like polyhydroxyalkanoates (PHA) are the best natural macromolecules to use as alternative to the synthetic polymers. Many prokaryotes accumulate PHA as cytoplasmic intracellular granules and their accumulation is triggered by starving conditions. The PHAs are ecofriendly and used to create biodegradable plastics. The microbial synthesized PHA had acquired global importance in industrial and biomedical sectors.</p><p><strong>Results: </strong>Ten different bacterial strains were isolated for the screening of PHA producers from the estuarine region of the Bay of Bengal, Suryalanka in Bapatla. A yellowish slimy circular colony known as M4 is actively growing on selective minimal media and was screened for polymeric granules in its cytoplasm using Sudan Black B and confirmed with the fluorescent dye Nile blue A. All of the isolates were biochemically tested and isolate M4 is the most capable of growing at high NaCl concentrations (3.2 percent) and tests positive for catalase, methyl red. The M4 strain revealed clear hydrolysis of gelatin, starch, and casein. The 16S rRNA sequencing revealed that M4 is 99.72% of identity to Brachybacterium paraconglomeratum LMG 19861(T) in BLAST and the obtained strain was assigned with accession no. MTCC 13074 and deposited in NCBI with accession no. MW899045. The chief cellular fatty acids found in M4 were C14:0, C15:0, C16:0, C18:1cis-9, C18:0, iso-C15: 0, iso-C14: 0, anteiso-C17: 0 and C18:1-7. Crotonic acid formation from M4-PHB extract was detected at 235nm in a UV spectrophotometer. Methanolysis was done, and derivatives of polyhydroxybutyric acid (PHB) in the extract were analyzed using GC-MS. Increasing viscosity was seen in the extracts which confirms the presence of polymer in the extracts. Thermogravimetric analysis was studied to determine the thermal profile of the PHB in the extract of M4.</p><p><strong>Conclusion: </strong>In the study, the selective screening and extraction of ecofriendly PHB from M4 strain was highlighted. Brachybacterium paraconglomeratum is a novel strain showed its uniqueness by producing few monomeric derivatives of PHB. The strain was reporting for the first time as PHA producer. B. paraconglomeratum has promising characteristics according to its metabolic profile. In addition, this study also helps to understand the diversity of bacteria isolated from marine sources.</p>","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":"21 1","pages":"24"},"PeriodicalIF":0.0,"publicationDate":"2023-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9975135/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10822973","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimization of recombinant antibody production based on the vector design and the level of metabolites for generation of Ig- producing stable cell lines.","authors":"V A Toporova,&nbsp;V V Argentova,&nbsp;T K Aliev,&nbsp;A A Panina,&nbsp;D A Dolgikh,&nbsp;M P Kirpichnikov","doi":"10.1186/s43141-023-00474-0","DOIUrl":"https://doi.org/10.1186/s43141-023-00474-0","url":null,"abstract":"<p><strong>Background: </strong>The biopharmaceutical industry is significantly growing worldwide, and the Chinese hamster ovary (CHO) cells are used as a main expression host for the production of recombinant monoclonal antibodies. Various metabolic engineering approaches have been investigated to generate cell lines with improved metabolic characteristics for increasing longevity and mAb production. A novel cell culture method based on the 2-stage selection makes it possible to develop a stable cell line with high-quality mAb production.</p><p><strong>Results: </strong>We have constructed several design options of mammalian expression vectors for the high production of recombinant human IgG antibodies. Versions for bipromoter and bicistronic expression plasmids different in promoter orientation and cistron arrangements were generated. The aim of the work presented here was to assess a high-throughput mAb production system that integrates the advantages of high-efficiency cloning and stable cell clones to stage strategy selection reducing the time and effort required to express therapeutic monoclonal mAbs. Development of a stable cell line using bicistronic construct with EMCV IRES-long link gave an advantage in high mAb expression and long-term stability. Two-stage selection strategies allowed the elimination of low-producer clones by using metabolic level intensity to estimate the IgG production in the early steps of selection. The practical application of the new method allows to reduce time and costs during stable cell line development.</p>","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":"21 1","pages":"23"},"PeriodicalIF":0.0,"publicationDate":"2023-02-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9947203/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9320079","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}