Microbial Biotechnology最新文献_第9页

De novo biosynthesis and nicotinamide biotransformation of nicotinamide mononucleotide by engineered yeast cells 工程酵母细胞对烟酰胺单核苷酸的新生物合成和烟酰胺生物转化。

IF 5.7 2区生物学

Microbial Biotechnology Pub Date : 2024-11-09 DOI: 10.1111/1751-7915.70048

Yanna Ren, Bei Han, Shijie Wang, Xingbin Wang, Qi Liu, Menghao Cai

{"title":"De novo biosynthesis and nicotinamide biotransformation of nicotinamide mononucleotide by engineered yeast cells","authors":"Yanna Ren, Bei Han, Shijie Wang, Xingbin Wang, Qi Liu, Menghao Cai","doi":"10.1111/1751-7915.70048","DOIUrl":"10.1111/1751-7915.70048","url":null,"abstract":"β-Nicotinamide mononucleotide (NMN) is a precursor of NAD+ in mammals. Research on NAD+ has demonstrated its crucial role against aging and disease. Here two technical paths were established for the efficient synthesis of NMN in the yeast Pichia pastoris, enabling the production of NMN from the low-cost nicotinamide (NAM) or basic carbon sources. The yeast host was systematically modified to adapt to the biosynthesis and accumulation of NMN. To improve the semi-biosynthesis of NMN from NAM, nicotinamide phosphoribosyltransferases were expressed intracellular to evaluate their catalytic activities. The accumulation of extracellular NMN was further increased by the co-expression of an NMN transporter. Fine-tuning of gene expression level produced 72.1 mg/L NMN from NAM in flasks. To achieve de novo biosynthesis NMN, a heterologous biosynthetic pathway was reassembled in yeast cells. Fine-tuning of pathway nodes by the modification of gene expression level and enhancement of precursor generation allowed efficient NMN synthesis from glucose (36.9 mg/L) or ethanol (57.8 mg/L) in flask. Lastly, cultivations in a bioreactor in fed-batch mode achieved an NMN titre of 1004.6 mg/L at 165 h from 2 g NAM and 868 g glucose and 980.4 mg/L at 91 h from 160 g glucose and 557 g ethanol respectively. This study provides a foundation for future optimization of NMN biosynthesis by engineered yeast cell factories.","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"17 11","pages":""},"PeriodicalIF":5.7,"publicationDate":"2024-11-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11549688/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142613274","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Enhanced biosynthesis of poly(3-hydroxybutyrate) in engineered strains of Pseudomonas putida via increased malonyl-CoA availability 通过增加丙二酰-CoA 的供应量，提高假单胞菌工程菌株中聚（3-羟基丁酸）的生物合成能力。

IF 5.7 2区生物学

Microbial Biotechnology Pub Date : 2024-11-06 DOI: 10.1111/1751-7915.70044

Giusi Favoino, Nicolas Krink, Tobias Schwanemann, Nick Wierckx, Pablo I. Nikel

{"title":"Enhanced biosynthesis of poly(3-hydroxybutyrate) in engineered strains of Pseudomonas putida via increased malonyl-CoA availability","authors":"Giusi Favoino, Nicolas Krink, Tobias Schwanemann, Nick Wierckx, Pablo I. Nikel","doi":"10.1111/1751-7915.70044","DOIUrl":"10.1111/1751-7915.70044","url":null,"abstract":"Malonyl-coenzyme A (CoA) is a key precursor for the biosynthesis of multiple value-added compounds by microbial cell factories, including polyketides, carboxylic acids, biofuels, and polyhydroxyalkanoates. Owing to its role as a metabolic hub, malonyl-CoA availability is limited by competition in several essential metabolic pathways. To address this limitation, we modified a genome-reduced Pseudomonas putida strain to increase acetyl-CoA carboxylation while limiting malonyl-CoA utilization. Genes involved in sugar catabolism and its regulation, the tricarboxylic acid (TCA) cycle, and fatty acid biosynthesis were knocked-out in specific combinations towards increasing the malonyl-CoA pool. An enzyme-coupled biosensor, based on the rppA gene, was employed to monitor malonyl-CoA levels in vivo. RppA is a type III polyketide synthase that converts malonyl-CoA into flaviolin, a red-colored polyketide. We isolated strains displaying enhanced malonyl-CoA availability via a colorimetric screening method based on the RppA-dependent red pigmentation; direct flaviolin quantification identified four engineered strains had a significant increase in malonyl-CoA levels. We further modified these strains by adding a non-canonical pathway that uses malonyl-CoA as precursor for poly(3-hydroxybutyrate) biosynthesis. These manipulations led to increased polymer accumulation in the fully engineered strains, validating our general strategy to boost the output of malonyl-CoA–dependent pathways in P. putida.","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"17 11","pages":""},"PeriodicalIF":5.7,"publicationDate":"2024-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11539682/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142581547","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Genome reduction improves octanoic acid production in scale down bioreactors 减少基因组可提高缩比生物反应器中辛酸的产量。

IF 5.7 2区生物学

Microbial Biotechnology Pub Date : 2024-11-06 DOI: 10.1111/1751-7915.70034

William T. Cordell, Gennaro Avolio, Ralf Takors, Brian F. Pfleger

{"title":"Genome reduction improves octanoic acid production in scale down bioreactors","authors":"William T. Cordell, Gennaro Avolio, Ralf Takors, Brian F. Pfleger","doi":"10.1111/1751-7915.70034","DOIUrl":"10.1111/1751-7915.70034","url":null,"abstract":"Microorganisms in large-scale bioreactors are exposed to heterogeneous environmental conditions due to physical mixing constraints. Nutritional gradients can lead to transient expression of energetically wasteful stress responses and as a result, can reduce the titres, rates and yields of a bioprocess at larger scales. To what extent these process parameters are impacted is often unknown and therefore bioprocess scale-up comes with major risk. Designing platform strains to account for these intermittent stresses before introducing synthesis pathways is one strategy for de-risking bioprocess development. For example, Escherichia coli strain RM214 is a derivative of wild-type MG1655 that has had several genes and whole operons removed from its genome based on their metabolic cost. In this study, we engineered E. coli strain RM214 (referred to as WG02) to produce octanoic acid from glycerol in batch-flask and fed-batch bioreactor cultivations and compared it to an octanoic acid-producing E. coli MG1655 (WG01). In batch flask cultivations, the two strains performed similarly. However, in carbon limited fed-batch bioreactor cultivations, WG02 provided a greater than 22% boost to biomass compared to WG01 while maintaining similar titres of octanoic acid. Reducing the biomass accumulation of WG02 with nitrogen limited fed-batch cultivation resulted in a 16% improvement in octanoic acid titre over WG01. Finally, in a scale-down system consisting of a stirred tank reactor (representing a well-mixed zone) and plug flow reactor (representing an intermittent carbon starvation zone), WG02 again improved octanoic acid titre by almost 18% while maintaining similar biomass concentrations as WG01.","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"17 11","pages":""},"PeriodicalIF":5.7,"publicationDate":"2024-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11540873/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142589944","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Thermodynamic constraints on the window of opportunity for direct interspecies electron transfer (DIET) 种间直接电子转移（DIET）机会窗口的热力学约束。

IF 5.7 2区生物学

Microbial Biotechnology Pub Date : 2024-11-02 DOI: 10.1111/1751-7915.70019

Jan Dolfing

引用次数: 0

Proton motive force and antibiotic tolerance in bacteria 质子动力与细菌的抗生素耐受性

IF 5.7 2区生物学

Microbial Biotechnology Pub Date : 2024-11-02 DOI: 10.1111/1751-7915.70042

Yingkun Wan, Jiaqi Zheng, Edward Wai-Chi Chan, Sheng Chen

{"title":"Proton motive force and antibiotic tolerance in bacteria","authors":"Yingkun Wan, Jiaqi Zheng, Edward Wai-Chi Chan, Sheng Chen","doi":"10.1111/1751-7915.70042","DOIUrl":"10.1111/1751-7915.70042","url":null,"abstract":"Bacterial antibiotic tolerance is a decades-old phenomenon in which a bacterial sub-population, commonly known as persisters, does not respond to antibiotics and remains viable upon prolonged antimicrobial treatment. Persisters are detectable in populations of bacterial strains that are not antibiotic-resistant and are known to be responsible for treatment failure and the occurrence of chronic and recurrent infection. The clinical significance of antibiotic tolerance is increasingly being recognized and comparable to antibiotic resistance. To eradicate persisters, it is necessary to understand the cellular mechanisms underlying tolerance development. Previous works showed that bacterial antibiotic tolerance was attributed to the reduction in metabolic activities and activation of the stringent response, SOS response and the toxin–antitoxin system which down-regulates transcription functions. The latest research findings, however, showed that decreased metabolic activities alone do not confer a long-lasting tolerance phenotype in persisters, and that active defence mechanisms such as efflux and DNA repair are required for the long-term maintenance of phenotypic tolerance. As such active tolerance-maintenance mechanisms are energy-demanding, persisters need to generate and maintain the transmembrane proton motive force (PMF) for oxidative phosphorylation. This minireview summarizes the current understanding of cellular mechanisms essential for prolonged expression of phenotypic antibiotic tolerance in bacteria, with an emphasis on the importance of generation and maintenance of PMF in enabling proper functioning of the active tolerance mechanisms in persisters. How such mechanisms can be utilized as targets for the development of anti-persister strategies will be discussed.","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"17 11","pages":""},"PeriodicalIF":5.7,"publicationDate":"2024-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70042","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142563402","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Harnessing Gram-negative bacteria for novel anti-Gram-negative antibiotics 利用革兰氏阴性菌开发新型抗革兰氏阴性菌抗生素。

IF 5.7 2区生物学

Microbial Biotechnology Pub Date : 2024-11-02 DOI: 10.1111/1751-7915.70032

Joy Birkelbach, Carsten E. Seyfert, Sebastian Walesch, Rolf Müller

引用次数: 0

AI in microbiome-related healthcare 人工智能在微生物相关医疗保健领域的应用。

IF 5.7 2区生物学

Microbial Biotechnology Pub Date : 2024-11-02 DOI: 10.1111/1751-7915.70027

Niklas Probul, Zihua Huang, Christina Caroline Saak, Jan Baumbach, Markus List

引用次数: 0

Metabolic engineering of Streptomyces roseosporus for increased production of clinically important antibiotic daptomycin 利用玫瑰孢链霉菌的代谢工程提高临床上重要的抗生素达托霉素的产量。

IF 5.7 2区生物学

Microbial Biotechnology Pub Date : 2024-11-02 DOI: 10.1111/1751-7915.70038

Xingwang Li, Ziwei Sang, Xuejin Zhao, Ying Wen

{"title":"Metabolic engineering of Streptomyces roseosporus for increased production of clinically important antibiotic daptomycin","authors":"Xingwang Li, Ziwei Sang, Xuejin Zhao, Ying Wen","doi":"10.1111/1751-7915.70038","DOIUrl":"10.1111/1751-7915.70038","url":null,"abstract":"Daptomycin (DAP), a novel cyclic lipopeptide antibiotic produced by Streptomyces roseosporus, is clinically important for treatment of infections caused by multidrug-resistant Gram-positive pathogens, but the low yield hampers its large-scale industrial production. Here, we describe a combination metabolic engineering strategy for constructing a DAP high-yielding strain. Initially, we enhanced aspartate (Asp) precursor supply in S. roseosporus wild-type (WT) strain by separately inhibiting Asp degradation and competitive pathway genes using CRISPRi and overexpressing Asp synthetic pathway genes using strong promoter kasOp*. The resulting strains all showed increased DAP titre. Combined inhibition of acsA4, pta, pyrB, and pyrC increased DAP titre to 167.4 μg/mL (73.5% higher than WT value). Co-overexpression of aspC, gdhA, ppc, and ecaA led to DAP titre 168 μg/mL (75.7% higher than WT value). Concurrently, we constructed a chassis strain favourable for DAP production by abolishing by-product production (i.e., deleting a 21.1 kb region of the red pigment biosynthetic gene cluster (BGC)) and engineering the DAP BGC (i.e., replacing its native dptEp with kasOp*). Titre for the resulting chassis strain reached 185.8 μg/mL. Application of our Asp precursor supply strategies to the chassis strain further increased DAP titre to 302 μg/mL (2.1-fold higher than WT value). Subsequently, we cloned the engineered DAP BGC and duplicated it in the chassis strain, leading to DAP titre 274.6 μg/mL. The above strategies, in combination, resulted in maximal DAP titre 350.7 μg/mL (2.6-fold higher than WT value), representing the highest reported DAP titre in shake-flask fermentation. These findings provide an efficient combination strategy for increasing DAP production and can also be readily applied in the overproduction of other Asp-related antibiotics.","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"17 11","pages":""},"PeriodicalIF":5.7,"publicationDate":"2024-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70038","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142563369","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Unlocking the potential for microbiome-based therapeutics to address the sustainable development goal of good health and wellbeing 释放微生物组疗法的潜力，实现良好健康和福祉的可持续发展目标。

IF 5.7 2区生物学

Microbial Biotechnology Pub Date : 2024-11-02 DOI: 10.1111/1751-7915.70041

Emily L. Gulliver, Sara K. Di Simone, Michelle Chonwerawong, Samuel C. Forster

{"title":"Unlocking the potential for microbiome-based therapeutics to address the sustainable development goal of good health and wellbeing","authors":"Emily L. Gulliver, Sara K. Di Simone, Michelle Chonwerawong, Samuel C. Forster","doi":"10.1111/1751-7915.70041","DOIUrl":"10.1111/1751-7915.70041","url":null,"abstract":"Recent years have witnessed major advances and an ever-growing list of healthcare applications for microbiome-based therapeutics. However, these advances have disproportionately targeted diseases common in high-income countries (HICs). Within low- to middle-income countries (LMIC), opportunities for microbiome-based therapeutics include sexual health epidemics, maternal health, early life mortality, malnutrition, vaccine response and infectious diseases. In this review we detail the advances that have been achieved in microbiome-based therapeutics for these areas of healthcare and identify where further work is required. Current efforts to characterise microbiomes from LMICs will aid in targeting and optimisation of therapeutics and preventative strategies specifically suited to the unmet needs within these populations. Once achieved, opportunities from disease treatment and improved treatment efficacy through to disease prevention and vector control can be effectively addressed using probiotics and live biotherapeutics. Together these strategies have the potential to increase individual health, overcome logistical challenges and reduce overall medical, individual, societal and economic costs.","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"17 11","pages":""},"PeriodicalIF":5.7,"publicationDate":"2024-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70041","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142563404","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Optimized psilocybin production in tryptophan catabolism-repressed fungi 优化色氨酸分解抑制真菌的迷幻药生产。

IF 5.7 2区生物学

Microbial Biotechnology Pub Date : 2024-11-02 DOI: 10.1111/1751-7915.70039

Slavica Janevska, Sophie Weiser, Ying Huang, Jun Lin, Sandra Hoefgen, Katarina Jojić, Amelia E. Barber, Tim Schäfer, Janis Fricke, Dirk Hoffmeister, Lars Regestein, Vito Valiante, Johann E. Kufs

{"title":"Optimized psilocybin production in tryptophan catabolism-repressed fungi","authors":"Slavica Janevska, Sophie Weiser, Ying Huang, Jun Lin, Sandra Hoefgen, Katarina Jojić, Amelia E. Barber, Tim Schäfer, Janis Fricke, Dirk Hoffmeister, Lars Regestein, Vito Valiante, Johann E. Kufs","doi":"10.1111/1751-7915.70039","DOIUrl":"10.1111/1751-7915.70039","url":null,"abstract":"The high therapeutic potential of psilocybin, a prodrug of the psychotropic psilocin, holds great promise for the treatment of mental disorders such as therapy-refractory depression, alcohol use disorder and anorexia nervosa. Psilocybin has been designated a ‘Breakthrough Therapy’ by the US Food and Drug Administration, and therefore a sustainable production process must be established to meet future market demands. Here, we present the development of an in vivo psilocybin production chassis based on repression of l-tryptophan catabolism. We demonstrate the proof of principle in Saccharomyces cerevisiae expressing the psilocybin biosynthetic genes. Deletion of the two aminotransferase genes ARO8/9 and the indoleamine 2,3-dioxygenase gene BNA2 yielded a fivefold increase of psilocybin titre. We transferred this knowledge to the filamentous fungus Aspergillus nidulans and identified functional ARO8/9 orthologs involved in fungal l-tryptophan catabolism by genome mining and cross-complementation. The double deletion mutant of A. nidulans resulted in a 10-fold increased psilocybin production. Process optimization based on respiratory activity measurements led to a final psilocybin titre of 267 mg/L in batch cultures with a space–time-yield of 3.7 mg/L/h. These results demonstrate the suitability of our engineered A. nidulans to serve as a production strain for psilocybin and other tryptamine-derived pharmaceuticals.","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"17 11","pages":""},"PeriodicalIF":5.7,"publicationDate":"2024-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70039","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142563401","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0