Chunsheng Yan, Ikechukwu C Okorafor, Colin W Johnson, Kendall N Houk, Neil K Garg, Yi Tang
{"title":"Microbial Biosynthesis of Rare Cannabinoids.","authors":"Chunsheng Yan, Ikechukwu C Okorafor, Colin W Johnson, Kendall N Houk, Neil K Garg, Yi Tang","doi":"10.1093/jimb/kuaf013","DOIUrl":"https://doi.org/10.1093/jimb/kuaf013","url":null,"abstract":"<p><p>∆9-tetrahydrocannabinol (∆9-THC) and cannabidiol (CBD) are the most abundant natural cannabinoids isolated from the different cultivars of the Cannabis plant. Other natural ∆9-THC analogs, especially those with different alkyl chain substitutions, display different and potent bioactivity. However, these rare cannabinoids are typically isolated at minuscule amounts and are difficult to synthesize. Targeted microbial biosynthesis can therefore be an attractive route to access such molecules. Here, we report the development of a Saccharomyces cerevisiae host to biosynthesize two rare cannabinoids from simple sugars. The yeast host is engineered to accumulate excess geranyl pyrophosphate (GPP), to overexpress a fungal pathway to 2,4-dihydroxy-6-alkyl-benzoic acids, as well as the downstream UbiA-prenyltransferase and THCA synthase. Two rare cannabinoid acids, the C1-substituted ∆9-tetrahydrocannabiorcolic acid (∆9-THCCA, ∼16 mg/L) and the C7-substituted ∆9-tetrahydrocannabiphorolic acid (∆9-THCPA, ∼5 mg/L) were obtained from this host; the latter was thermally decarboxylated to give ∆9-tetrahydrocannabiphorol (∆9-THCP). Given the diversity of fungal biosynthetic gene clusters (BGCs) that can produce resorcylic acids, this microbial platform offers potential to produce other rare and new-to-nature cannabinoids.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143996141","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}
{"title":"Enhanced amino acid biosynthesis in Phaffia rhodozyma via herbicide-induced selection.","authors":"Svetlana Raita, Iveta Kuzmika, Taras Mika, Zane Geiba, Kriss Spalvins","doi":"10.1093/jimb/kuaf011","DOIUrl":"https://doi.org/10.1093/jimb/kuaf011","url":null,"abstract":"<p><p>According to the Food and Agricultural Organisation 2024 statement, developing single-cell protein technology is important to reduce the burden on conventional feed protein production sectors. In this regard, improved commercial strains rich in amino acids, especially Lys and Met, may provide a sustainable alternative source of protein in aquaculture diets. The developed and laboratory-validated methodology for creating protein-synthesising mutants will strengthen the competitiveness of SCP production technology. The present work provides unique results on improving the protein-producing properties of wild-type Phaffia rhodozyma DSM 5626 by mutagenesis and screening on herbicide-containing medium as a selective agent for amino acid biosynthesis inhibition. Inhibitory concentrations of pure herbicide actives were determined for S-(2-aminoethyl)-L-cysteine (AEC) and glufosinate-ammonium (GA) for complete inhibition and strong inhibition of the DSM 5626 strain. GA at a concentration of 50 mM and 100 mM and AEC at 0.5 mM and 2.5 mM were chosen for mutant selection after chemical mutagenesis. The use of herbicides resulted in the selection of mutants with significantly improved synthesis of Met and Lys. Specifically, mutants GA6/4 and GA7/5 exhibited 37% and 26% higher Met levels, respectively, while GA6/3 had a 14% increase in Lys compared to the wild-type strain. The AEC3/9 mutant demonstrated a 35% increase in Met, 24% in Lys, 8% in Ile, and 6% in Phe, underscoring the efficacy of this screening approach in enhancing essential amino acid content. The protein quality parameters EAAI and AAS of these mutants became higher compared with commercial strains of SCP yeast such as C. utilis, S. cerevisiae, K. marxianus, etc.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144021174","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}
{"title":"Microbial Enzymes in Industrial Biotechnology: Sources, Production and Significant Applications of Lipases.","authors":"Nisha Sharma, Yogesh K Ahlawat, Nattan Stalin, Sajid Mehmood, Sonia Morya, Anurag Malik, Malathi H, Jayshree Nellore, Deepak Bhanot","doi":"10.1093/jimb/kuaf010","DOIUrl":"https://doi.org/10.1093/jimb/kuaf010","url":null,"abstract":"<p><p>The variety of microorganisms represents the most prevalent sources utilized within diverse industries and research fields. Enzymes with microorganisms are applied in the use of industrial biotechnology. Since the dawn of civilization, there are techniques like extraction and fermentation that used plant or bacterial enzymes as well as other byproducts. Enzymes, the natural catalysts, are intricately involved in many aspects of life. Enzymes pose remarkable specificity for their substrate, which implies that these metabolic cycles in a living cell need to be executed by a team working in collaboration. The major sources of these enzymes are yeast, some fungi and bacteria. Just like all living forms, microbes interact with their environment in which they must live in order to survive. A large number of microorganisms that are capable of producing great varieties of enzymes are important in the production of bread, cheese, yogurt, beer, and many other foods. One of the most widely used lipolytic enzyme is lipase from various sources including food and dairy industry, leather, detergent, pulp and paper, bioenergy and even pharma. With the latest innovation in biotechnology, the need for organisms that produce different commercially important lipases which other strains of lipases do is increasing. Lipases produced from microbial cells have a major industrial significance because of their property versatility and ease of mass production. This review seeks to clarify the sources of microorganisms, lipase production and purification processes, as well as the environmental and industrial uses of lipase enzymes.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144016825","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}
Hyun Jin Kim, Yeda Lee, Yuni Shin, Suhye Choi, Jinok Oh, Suwon Kim, Jungoh Ahn, Kwon-Young Choi, Jeong Chan Joo, Shashi Kant Bhatia, Yung-Hun Yang
{"title":"Finding of the positive impact of glucose on the production of indican over indigo in engineered Escherichia coli.","authors":"Hyun Jin Kim, Yeda Lee, Yuni Shin, Suhye Choi, Jinok Oh, Suwon Kim, Jungoh Ahn, Kwon-Young Choi, Jeong Chan Joo, Shashi Kant Bhatia, Yung-Hun Yang","doi":"10.1093/jimb/kuae048","DOIUrl":"10.1093/jimb/kuae048","url":null,"abstract":"<p><p>Indigo is a plant-based natural blue dye that can be produced via chemical synthesis and biological pathways. However, the toxic reduction processes and intracellular production of indigo through microbial metabolism are often limited by insolubility of indigo and complex downstream processing, causing environmental issues in the dyeing processes. Additionally, indican, a precursor of indigo with a glucose moiety, is highly soluble and can be easily converted into indoxyl by β-glucosidase, forming indigo under mild conditions. We constructed an indican-producing strain Escherichia coli BL21 HI201 by introducing a UDP-glycosyltransferase (ugt) into an indoxyl production system containing tryptophanse (tnaA) and flavin-containing monooxygenase (FMO) genes, enabling conversion of tryptophan into indican. Testing of the effect by various carbon sources suggested that glucose is one of the major factors affecting the ratio of indigo to indican, and increase in glucose concentration to more than 1.5% could produce sole indican without indigo. Under optimal conditions, E. coli BL21 HI201 biosynthesized 5.65 mM indican from tryptophan. Additionally, after deletion of various β-glucosidase genes, the bglA knockout strain E. coli BL21 HI204 produced more indican, achieving 6.79 mM after 24 hr of cultivation. This study demonstrated the strategic production of indican through the installation of a production system, deletion of a byproduct pathway, and control of glucose concentration.</p><p><strong>One-sentence summary: </strong>This paper demonstrates the strategic enhancement of indican production in genetically engineered Escherichia coli BL21 by optimizing metabolic pathways and controlling glucose concentrations.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":"52 ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11947661/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143730374","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Caitlin A McCadden, Tyler A Alsup, Ion Ghiviriga, Jeffrey D Rudolf
{"title":"Biocatalytic diversification of abietic acid in Streptomyces.","authors":"Caitlin A McCadden, Tyler A Alsup, Ion Ghiviriga, Jeffrey D Rudolf","doi":"10.1093/jimb/kuaf003","DOIUrl":"10.1093/jimb/kuaf003","url":null,"abstract":"<p><p>Biocatalysis provides access to synthetically challenging molecules and commercially and pharmaceutically relevant natural product analogs while adhering to principles of green chemistry. Cytochromes P450 (P450s) are among the most superlative and versatile oxidative enzymes found in nature and are desired regio- and stereoselective biocatalysts, particularly for structurally complex hydrocarbon skeletons. We used 10 genome-sequenced Streptomyces strains, selected based on their preponderance of P450s, to biotransform the bioactive diterpenoid abietic acid. We isolated and structurally characterized seven oxidized abietic acid derivatives from three different strains, including four products that are new bacterial biotransformants or enzymatic products. Oxidations (hydroxylation, dehydrogenation, and aromatization) were seen on both the B and C rings of abietic acid and five products had multiple modifications. Notable conversions observed in the study were that of abietic acid to 15-hydroxy-7-oxo-8,11,13-abietatrien-18-oic acid, 7, which involves multiple hydroxylation reactions and dehydrogenation. The findings from this study will lead to identifying P450s or other enzymes that may act as general biocatalysts to modify abietanes and other labdane-type diterpenoid skeletons.</p><p><strong>One-sentence summary: </strong>Genome-guided biotransformation of the bioactive diterpenoid abietic acid in Streptomyces yielded seven oxidized derivatives including four that have not been previously seen from bacteria.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11812575/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143065574","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ambreen Latif, Noor Hassan, Hazrat Ali, Muhammad Bilal Khan Niazi, Zaib Jahan, Iqra Latif Ghuman, Farwa Hassan, Anam Saqib
{"title":"An overview of key industrial product citric acid production by Aspergillus niger and its application.","authors":"Ambreen Latif, Noor Hassan, Hazrat Ali, Muhammad Bilal Khan Niazi, Zaib Jahan, Iqra Latif Ghuman, Farwa Hassan, Anam Saqib","doi":"10.1093/jimb/kuaf007","DOIUrl":"10.1093/jimb/kuaf007","url":null,"abstract":"<p><p>Citric acid possesses high economic value and is considered as the world's largest consumed organic acid in numerous industries. Citric acid applications range from food to beverage industries, pharmaceuticals, cosmetics, and the environment. It is mostly produced by microbial fermentation, but Aspergillus niger is considered as the main workhorse for large-scale production of citric acid. In the current review, special devotion has been made toward addressing the latest and innovative literature related to production of citric acid by A. niger. The review article discusses A. niger historical involvement in citric acid production, fermentation technologies, molecular biology, biosynthesis, accumulation of citric acid, methods for enhanced production of citric acid, different operational factors also influencing citric acid production, and various techniques used for citric acid recovery. Also, copious biotechnological applications of citric acid are summarized for a fundamental comprehension of the subject and its critical role in diverse fields of industries.</p><p><strong>One-sentence summary: </strong>This review describes the historical role of Aspergillus niger in the production of citric acid, fermentation technologies, molecular biology, techniques for increased citric acid production, and other physical and chemical variables influencing the production of citric acid.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11956825/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143742898","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Amr A Hemeda, Sara A Zahran, Marwa Ali-Tammam, Menna A Ewida, Mona T Kashef, Aymen S Yassin, Avishek Mitra, Noha H Youssef, Mostafa S Elshahed
{"title":"Metagenomic mining unveils a novel GH130 enzyme with exclusive xylanase activity over a wide temperature and pH ranges.","authors":"Amr A Hemeda, Sara A Zahran, Marwa Ali-Tammam, Menna A Ewida, Mona T Kashef, Aymen S Yassin, Avishek Mitra, Noha H Youssef, Mostafa S Elshahed","doi":"10.1093/jimb/kuaf006","DOIUrl":"10.1093/jimb/kuaf006","url":null,"abstract":"<p><p>The equine gut harbors a diverse microbial community and represents a rich source of carbohydrate-active enzymes (CAZymes). To identify and characterize potentially novel CAZymes from a horse's hindgut metagenome, shotgun metagenomic sequencing was performed on DNA extracted from a stool sample of a male horse, followed by CAZyme annotation. Here, we report on the characterization of a novel enzyme (AH2) that was identified, synthesized, cloned, and characterized from the obtained CAZyme dataset. AH2 was identified as a GH130 family member and displayed exclusive xylanase activity, a trait hitherto unreported in prior characterization of GH130 CAZymes. AH2 displayed an optimal activity at a pH of 5.6 and a temperature of 50°C. AH2 maintained significant activity across a pH range of 4-10 (62-72%) and temperatures of 30-70°C (77-86%). The enzyme had remarkable stability, with minimal reductions in activity across a temperature range of 4-70°C and pH levels of 3, 7, and 9. Docking studies identified AH2's amino acids (Glu90 and Glu149) to be involved in substrate binding. Molecular dynamics simulation confirmed the structural stability of AH2 at pH 5.6 and 50°C, further supporting its resilience under these conditions. Our results expand on the known activities associated with the GH130 CAZyme family and demonstrate that the horse gut metagenome represents an unexplored source of novel CAZymes.</p><p><strong>One-sentence summary: </strong>A novel activity for members of the CAZyme family GH130.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11905756/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143557068","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ibrahim M Elgendy, Nehal E Elkaliny, Hoda M Saleh, Gehad O Darwish, Mervt M Almostafa, Kamel Metwally, Galal Yahya, Yehia A-G Mahmoud
{"title":"Bacteria-powered self-healing concrete: Breakthroughs, challenges, and future prospects.","authors":"Ibrahim M Elgendy, Nehal E Elkaliny, Hoda M Saleh, Gehad O Darwish, Mervt M Almostafa, Kamel Metwally, Galal Yahya, Yehia A-G Mahmoud","doi":"10.1093/jimb/kuae051","DOIUrl":"10.1093/jimb/kuae051","url":null,"abstract":"<p><p>In a world where concrete structures face constant degradation from environmental forces, a revolutionary solution has emerged: bio-self-healing concrete. This innovation involves embedding dormant bacteria within the concrete mix, poised to spring into action when cracks form. As moisture seeps into the cracks, these bacterial agents are activated, consuming nutrients and converting them into calcium carbonate, a natural substance that fills and repairs the fractures, restoring the material's integrity. This fascinating process represents a cutting-edge approach to maintaining concrete infrastructure, turning once-vulnerable materials into self-sustaining systems capable of healing themselves. The ongoing research into bio-self-healing concrete is focused on selecting bacterial strains that can withstand the extreme conditions within concrete, including its highly alkaline environment. The bacteria must also form resilient spores, remaining viable until they are needed for repair. Additionally, the study explores various challenges associated with this technology, such as the cost of production, the bacteria's long-term viability, and their potential environmental impact. Advancements in genetic engineering and smart technology are being explored to enhance these bacterial strains, making them more efficient and robust in their role as microscopic repair agents. This review delves into the potential of bio-self-healing concrete to revolutionize how we approach infrastructure maintenance, offering a glimpse into a future where concrete structures not only endure but actively repair themselves, extending their lifespan and reducing the need for costly repairs.</p><p><strong>One-sentence summary: </strong>Bio-self-healing concrete utilizes bacteria that activate upon crack formation to repair structures by producing calcium carbonate, offering a sustainable solution to prolong the lifespan of concrete infrastructure.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11730074/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142824235","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Navindu Dinara Gajanayaka, Eunyoung Jo, Minthari Sakethanika Bandara, Svini Dileepa Marasinghe, Sachithra Amarin Hettiarachchi, Sithumini Wijewickrama, Gun-Hoo Park, Chulhong Oh, Youngdeuk Lee
{"title":"Pseudoalteromonas agarivorans-derived novel ulvan lyase of polysaccharide lyase family 40: Potential application of ulvan and partially hydrolyzed products in cosmetic industry.","authors":"Navindu Dinara Gajanayaka, Eunyoung Jo, Minthari Sakethanika Bandara, Svini Dileepa Marasinghe, Sachithra Amarin Hettiarachchi, Sithumini Wijewickrama, Gun-Hoo Park, Chulhong Oh, Youngdeuk Lee","doi":"10.1093/jimb/kuaf004","DOIUrl":"10.1093/jimb/kuaf004","url":null,"abstract":"<p><p>Ulvan is a complex sulfated polysaccharide in the cell walls of green algae with extensive applications in food, pharmaceutical, and agricultural industries, prompting extensive studies on ulvan, its oligosaccharides, monosaccharides, and cost-effective depolymerization methods. Our primary objectives were to investigate novel ulvan-utilizing marine bacteria, perform recombinant engineering of genes responsible for ulvan depolymerization, and determine their potential industrial applications. Samples were collected from Jeju Island, which is a South Korean region with significant excessive green algal growth, especially that of Ulva species. The marine bacterium Pseudoalteromonas agarivorans efficiently uses ulvan as its primary carbon source, indicating its potential for ulvan degradation. Through whole-genome sequencing the paul40 gene, which is a polysaccharide lyase family 40 (PL40) member, was identified and subsequently engineered into the pET-16b vector for expression as a His-tagged 95 kDa fusion protein. The ulvan depolymerization process was evaluated and confirmed using various analytical techniques including dinitrosalicylic acid assay, thin-layer chromatography, and gel permeation chromatography. Optimal enzyme activity occurred at 35°C, pH 8.0 in phosphate buffer, and 2.5 mM of NaCl. Furthermore, enzyme characterization and specific activity measurements were performed. This study is the first to report hyaluronidase and elastase inhibition by ulvan and its derivatives along with the characterization of an ulvan lyase enzyme from the PL40 family.</p><p><strong>One-sentence summary: </strong>This study reports the identification and recombinant expression of a novel ulvan-degrading enzyme from Pseudoalteromonas agarivorans, demonstrating its potential for cosmetic industrial applications by revealing ulvan's and partially hydrolyzed ulvan's hyaluronidase and elastase inhibition properties.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11835017/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143370648","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Jennifer Reid, Joyce Ni, Airong Chen, Patricia Gomes, Andrew Szto, Analyn Yu, Angela Luo, Belinda Kong, Calvin Adams, Neveathan Jeyachandran, Anumta Amir, Xavier Teixeira, Tao Yuan, Cédric Charretier
{"title":"Exploration of alternative microfiltration modalities for the harvest and clarification of diverse recombinant proteins from high-density E. coli culture and lysate using hollow fibre, flat sheet cassette, and vibro membrane filtration technologies.","authors":"Jennifer Reid, Joyce Ni, Airong Chen, Patricia Gomes, Andrew Szto, Analyn Yu, Angela Luo, Belinda Kong, Calvin Adams, Neveathan Jeyachandran, Anumta Amir, Xavier Teixeira, Tao Yuan, Cédric Charretier","doi":"10.1093/jimb/kuaf008","DOIUrl":"10.1093/jimb/kuaf008","url":null,"abstract":"<p><p>Industrial bioprocess optimization has significantly increased the productivity of biomass and biologics in upstream production. Such process improvement in fermentation often translates to challenges in recovering intracellularly expressed recombinant proteins due to increased matrix complexity, resulting in a higher performance burden in midstream. Tangential flow filtration (TFF) is a popular industry standard for buffer exchange and protein separation from cellular debris. However, due to variations in the physicochemical properties of recombinant proteins, solutions for E. coli-based protein clarification remain challenging and often necessitate extensive exploration and process optimization. With growing options in filtration-based technologies, the identification of a near-universal clarification platform is desirable to accelerate bioprocess development overall. In this study, three TFF modalities, hollow fibre (HF), flat-sheet cassette (CAS), and vibro membrane filtration (VMF), were assessed in parallel to evaluate their clarification performance for three E. coli recombinant proteins with different biochemical properties. Reverse phase liquid chromatography data showed target protein recovery was uniformly higher for VMF than HF at equivalent loading. Cell density and lysate protein load were comparable for HF and VMF, and lower for CAS. These results support the choice of VMF and HF as easily optimized and operated TFF modalities for clarification of recombinant protein from complex crude bacterial matrix, where either can be efficiently performed with ease and minimum supervision. Both TFF applications were successfully demonstrated in primary cell harvest, cell wash and cell lysate clarification, for E. coli-based recombinant proteins.</p><p><strong>One-sentence summary: </strong>High-density E. coli microfiltration and lysate clarification were tested for three diverse recombinant proteins, where hollow fibre and vibro membrane filtration outperformed flat sheet cassette in terms of process time, suspended solid loading, and target protein recovery.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12022607/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143788516","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}