Zhuhai Gene-Biocon Biological Technology Co., Ltd.’s Post

Recombinant Enzymes in Genetic Engineering: An Overview. In #genetic #engineering, recombinant #enzymes serve as the molecular scissors that precisely cut and rearrange DNA sequences. The most widely used enzyme in this process is the DNA restriction endonuclease. This versatile tool allows scientists to selectively cleave DNA at specific recognition sites, facilitating the manipulation of genes. Another essential enzyme in the genetic engineering toolkit is DNA ligase, which efficiently seals the gaps created during DNA splicing. Together, these enzymes enable the creation of genetically modified #organisms and pave the way for countless scientific breakthroughs. #biology #biological #bioscience #bioengineering #geneticengineering #biopharmaceuticals #biopharma #proteins https://lnkd.in/gcBNWznN

Compared with other expression systems, the Pichia pastoris expression system has the following advantages: (1) Contains a unique and powerful AOX (alcohol oxidase gene) promoter, which can strictly control the expression of foreign genes with methanol; (2) The expression level is high, and it can be expressed intracellularly or secretedly. In Pichia pastoris, the highest reported expression level of tetanus toxin C is 12g/l, which is generally greater than 1g/l. Most foreign genes are expressed at higher levels than in bacteria, Saccharomyces cerevisiae, and animal cells. Generally, exogenous genes in Pichia pastoris contain signal peptide sequences that guide secretion, so that the expressed exogenous target protein can be secreted into the fermentation broth, which is beneficial to isolation and purification; (3) The fermentation process is mature and easy to scale up. There is already a fermentation process for large-scale industrial high-density production, and the dry cell weight reaches more than 100g/l. When expressing recombinant proteins, it has been successfully scaled up to 10,000 liters; (4) The cultivation cost is low and the products are easy to separate. The fermentation medium used by Pichia pastoris is very cheap. Generally, the carbon sources are glycerol or glucose and methanol, and the rest are inorganic salts. The medium does not contain protein, which is conducive to the isolation and purification of downstream products; while the inducers used by Saccharomyces cerevisiae are generally relatively expensive. of galactose; (5) The foreign protein gene is genetically stable. Generally, foreign protein genes are integrated into the Pichia pastoris chromosome, replicated along with chromosome replication, and are not easily lost; (6) As a eukaryotic expression system, Pichia pastoris has a eukaryotic subcellular structure and has post-translational modification processing functions such as glycosylation, fatty acylation, and protein phosphorylation. #cell #biology #biological #Biotechnology #bioengineering #geneticengineering #expression https://lnkd.in/gcBNWznN

Understanding Recombinant Enzymes and Their Significance. Recombinant #enzymes are derived from genetically modified organisms (GMOs) and possess the ability to cut splice join, and replicate DNA molecules with remarkable precision. By harnessing these enzymes, scientists can manipulate genetic material with unprecedented accuracy, amplifying their impact on various industries. From medical research to agriculture, recombinant enzymes have paved the way for groundbreaking advancements. #enzymes #biology #biological #biosciences #geneticengineering #bioengineering #biopharmaceutical #biomedical https://lnkd.in/gcBNWznN

Promotion: LAB-A-PORTER x OriGene. Use Promo Codes To Redeem Your Offer Today! To support your research needs on recombinant proteins, OriGene and LAB-A-PORTER are thrilled to announce a special promotion for selected recombinant proteins! From now until 31 March 2024, for purchase of MVPro recombinant proteins from OriGene, you can receive 20% off. Limited offer. Grab the chance to stock up on some recombinant proteins! Use promo code: MVPRO20 to redeem your offer today! T&C applies. All products listed on this site and sold through LAB-A-PORTER Limited are for research use only (RUO). Not for human use. Not for therapeutic or diagnostic use. Learn more: https://lnkd.in/gbaewUMy

How to improve transient protein expression agroinfiltrated Nicotiana benthamiana?   Proteins (high-value proteins, including biopharmaceuticals, antibodies, antigens, enzymes, and vaccines) are expressed in large scale in Tobacco plants. This is important for plant science and molecular pharming/farming. Advantages: sustainability and cost-effectiveness   4 Strategies to enhance the efficiency of transient protein expression using agroinfiltration.   - Avoiding Immune Responses: Immune responses can hinder protein expression. Strategies include depleting immune receptors like NbCORE and using pathogen-derived effectors such as AvrPto to suppress immune responses, increasing protein yields.   - Avoiding Transcript Degradation: Transcripts can become unstable due to post-transcriptional gene silencing (PTGS). Using silencing inhibitors like P19 and mutant lines deficient in PTGS machinery (e.g., dcl2dcl4 mutants) enhances transcript stability and protein expression.   - Avoiding Endoplasmic Reticulum (ER) Stress: ER stress from unfolded proteins can be mitigated by co-expressing chaperones. Co-expressing human calreticulin has significantly improved the expression of proteins such as HIV glycoprotein gp140 and SARS-CoV-2 spike protein by enhancing protein folding and reducing ER stress.   - Avoiding Proteolysis: Proteolysis of recombinant proteins is a challenge. Strategies include co-expressing protease inhibitors (e.g., SlCYS8), silencing or knocking out specific proteases, and targeting proteins to different subcellular locations to evade degradation.   Future prospects include improving co-expression strategies, engineering post-translational modifications, optimizing plant growth conditions, and refining the protein extraction process. These advancements will make agroinfiltration a more robust and efficient platform for transient protein expression in plants.   #LeafExpressionsystems in the UK #IconGenetics in Germany acquired by Bayer #Tiamat Biosciences in the Research Triangle in North Carolina   Are there additional companies that use Tobacco for protein expression and molecular pharming? Please mention their names in the comments!   #PlantBiotechnology #MolecularPharming #MolecularFarming #ProteinExpression #Agroinfiltration #NicotianaBenthamiana #PlantScience

Introduction of Gene-Biocon Heat-stable Trypsin Solution. Gene-Biocon has launched a Heat-stable Trypsin Solution product currently. The product was named TrypRE select (1×). The TrypRE select (1×) is a recombinant thermostable trypsin digestion solution produced by fermentation, purification, and formulation from a genetically engineered strain of Pichia Pastoris. TrypRE select is a non-animal origin recombinant enzyme suitable for dissociating various adherent mammalian cells, including CHO, HEK 293, A529, primary human keratinocytes and embryonic stem cells. The recombinant enzyme used in this product is a trypsin-like protease from Fusarium oxysporum, a serine protease that can selectively hydrolyze the peptide chain composed of the carboxyl end of lysine or arginine in proteins, exhibiting enzymatic properties identical to those of recombinant porcine and bovine trypsin. #trypsin #recombinant #biologcial #biotechnology https://lnkd.in/gcBNWznN

Genetic engineering elements: Including exogenous DNA, vector molecules, tool enzymes and receptor cells, etc. The basic contents of a complete genetic engineering technology program for production purposes include: (1) Isolation and cloning of foreign target genes and study of the structure and function of target genes. This part of the work is the basis of the entire genetic engineering, so it is also called the upstream part of genetic engineering. (2) Suitable for transfer, construction of expression vectors or reorganization of expression control structures of target genes. (3) Introduction of foreign genes. (4) Integration, expression and detection of foreign genes on the host genome and screening of genetically modified organisms. (5) Verification of the physiological functions of exogenous gene expression products. (6) Breeding and establishment of new genetically modified lines, and analysis of benefits of new genetically modified lines. (7) Establishment of ecological and evolutionary security mechanisms. (8) Consumer safety evaluation. #geneticengineering #biology #biological #biotechnology #bioengineering https://lnkd.in/gcBNWznN

Identification of Inhibitors of Fungal Fatty Acid Biosynthesis Fungal fatty acid (FA) synthase and desaturase enzymes are essential for the growth and virulence of human fungal pathogens. These enzymes are structurally distinct from their mammalian counterparts, making them attractive targets for antifungal development. However, there has been little progress in identifying chemotypes that target fungal FA biosynthesis. To accomplish this, we applied a whole-cell-based method known as Target Abundance-based FItness Screening using Candida albicans. Strains with varying levels of FA synthase or desaturase expression were grown in competition to screen a custom small-molecule library. Hit compounds were defined as preferentially inhibiting the growth of the low target-expressing strains. Dose–response experiments confirmed that 16 hits (11 with an acyl hydrazide core) differentially inhibited the growth of strains with an altered desaturase expression, indicating a specific chemical–target interaction. Exogenous unsaturated FAs restored C. albicans growth in the presence of inhibitory concentrations of the most potent acyl hydrazides, further supporting the primary mechanism being inhibition of FA desaturase. A systematic analysis of the structure–activity relationship confirmed the acyl hydrazide core as essential for inhibitory activity. This collection demonstrated broad-spectrum activity against Candida auris and mucormycetes and retained the activity against azole-resistant candida isolates. Finally, a preliminary analysis of toxicity to mammalian cells identified potential lead compounds with desirable selectivities. Collectively, these results establish a scaffold that targets fungal FA biosynthesis with a potential for development into novel therapeutics. #drugdiscovery #drugdevelopment #candida

What is the future of microbial expression systems? 🦠 Microbial expression systems are widely used to manufacture diagnostic and therapeutic proteins. However, the commercialization success rate for new drug developments is less than 3%, partly due to high development and manufacturing costs, long turnaround times, and challenging product safety profiles. 💸 CDMOs, including KBI, are jumping in to update microbial expression systems to surpass the modest yields of traditional E. coli models. KBI Biopharma has developed PUREcoli, a microbial expression system incorporating over 1,000 gene edits to enhance protein production. 📈 Learn more about it in our latest article! ⬇️ https://lnkd.in/d-KT-xbg #biotechinnovation #proteinproduction #therapeuticproteins #CDMOs #drugdevelopment #biomanufacturing #biotechresearch #biotechindustry

📃Scientific paper: Identification of a major facilitator superfamily protein that is beneficial to L-lactic acid production by Bacillus coagulans at low pH Abstract: Background Product inhibition is one of the major problems in lactic acid (LA) fermentation. Our previous study revealed that Bacillus coagulans 2–6 was an efficient producer of high-optical-purity L-LA. Its mutant strain B. coagulans Na-2 has better resistance to sodium lactate stress but the resistance mechanism has not been understood. Results In this study, the whole-genome sequencing of B. coagulans Na-2 was performed and one mutant gene mfs coding for the major facilitator superfamily (MFS) protein was revealed by comparative genome analysis. Ten mutation sites were identified between the wild (MFS-2-6) and mutant (MFS-Na-2) proteins, among which T127A and N154T were predicted locating in the center of the transmembrane transport channel. The MFS-2-6 and MFS-Na-2 were expressed separately in a genetically operable strain, B. coagulans DSM1, using the genes’ native promoter. The expression of the two MFS proteins had no effect and a negative effect on L-LA production when the pH was controlled at 6.0 and 7.0 by sodium hydroxide, respectively. However, 4.2 and 4.6-fold of L-LA concentrations were obtained at pH 5.0 by the strains expressing MFS-2-6 and MFS-Na-2 than that by the control strain, respectively. The intracellular pH values of the strains expressing MFS-2-6 and MFS-Na-2 were approximately 0.69 and 0.45 higher than that of the control strain during pH-controlled fermentation at 5.0. Results suggest that the expression of MFS-2-6 and MFS-Na-2 were both... Continued on ES/IODE ➡️ https://etcse.fr/Cvd0 ------- If you find this interesting, feel free to follow, comment and share. We need your help to enhance our visibility, so that our platform continues to serve you.